specimen collection staining gastrointestinal respiratory urinary tract microbiology methods

https://www.pathologylab.org/filesimages/Microbiology%20Coll…

https://www.healthcare.uiowa.edu/path_handbook/appendix/micr…

staining methods gastrointestinal respiratory urinary tract specimens Gram Ziehl-Neelsen trichrome KOH microbiology

ls /home/daytona/workspace/

~/specimen-ppt/create_ppt.js

const pptxgen = require("pptxgenjs");

let pres = new pptxgen();
pres.layout = "LAYOUT_16x9";
pres.title = "Specimen Collection and Staining";
pres.author = "Orris";

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// ═════════════════════════════════════════════
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// SLIDE 2 – OVERVIEW / GENERAL PRINCIPLES
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    "Collect specimens BEFORE starting antimicrobial therapy whenever possible",
    "Use standard precautions — treat all specimens as potentially hazardous",
    "Label all containers: patient ID, date, time, collection site",
    "Deliver to laboratory as soon as possible after collection",
    "Transport guidelines: Urine — refrigerate within 30 min | Stool — within 1 hr | Respiratory — 2 hrs RT",
    "Viral specimens must be transported immediately on ice",
    "Do NOT use swabs from anaerobic flora-rich sites (expectorated sputum, voided urine) for anaerobic culture",
    "Specimen quality directly determines result quality — poor collection = unreliable results",
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// ═════════════════════════════════════════════
// SLIDE 3 – GI TRACT OVERVIEW
// ═════════════════════════════════════════════
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    ["Stool (feces)", "Pass into sterile wide-mouth leak-proof container; transfer from bedpan if needed. No toilet paper.", "Sterile screw-cap. Keep cool. Do NOT incubate."],
    ["Stool – O&P", "Same as above. Place in preservative immediately. 3 specimens over 7-10 days (not >1/day).", "Total-Fix / Protofix vial. RT."],
    ["Rectal Swab", "Insert swab 2-3 cm past anal sphincter; rotate to collect from rectal wall.", "Cary-Blair transport medium."],
    ["Gastric Biopsy / Lavage", "Endoscopic. Lavage → TB/H.pylori. Biopsy → 10% formalin (histo) or sterile saline (culture).", "Sterile container. Transport immediately."],
    ["Duodenal Aspirate", "Via endoscope or Enterotest (string test). For Giardia, microsporidia, Strongyloides.", "Sterile container on ice."],
    ["Sigmoidoscopy specimen", "Scrape/aspirate from mucosal lesions — NOT absorbent swabs. Multiple sites, process individually.", "Sterile container; prep wet smear immediately."],
    ["Pinworm (Enterobius)", "Clear adhesive tape to perianal area in morning BEFORE bathing/defecating. Stick to slide.", "Slide in protective case; pinworm collection kit."],
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// ═════════════════════════════════════════════
// SLIDE 4 – GI STAINING METHODS
// ═════════════════════════════════════════════
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    ["Wet Mount (direct)", "Motile protozoa (Giardia, Entamoeba), helminth eggs/larvae", "Saline ± iodine. Quick; requires fresh specimen."],
    ["Gram Stain", "Bacteria (Campylobacter curved rods), Candida, stool quality", "Gram+ = purple | Gram- = pink/red."],
    ["Trichrome (Wheatley)", "Entamoeba histolytica, Giardia, Blastocystis", "Background green; protozoa: blue-green/purple cytoplasm, red nuclei."],
    ["Iron Hematoxylin", "Intestinal protozoa (alternative to trichrome)", "Superior nuclear detail; more laborious."],
    ["Modified ZN (cold acid-fast)", "Cryptosporidium, Cyclospora, Cystoisospora (Isospora)", "Oocysts: pink-red on blue. No heating required."],
    ["Modified Trichrome Blue (Ryan)", "Microsporidia (E. bieneusi, E. intestinalis)", "Spores: pink-reddish. Also useful on sputum and urine."],
    ["Calcofluor White", "Microsporidia, fungi, Acanthamoeba cysts", "Fluorescence microscope needed. Binds chitin."],
    ["KOH", "Fungi: Candida, molds (esophageal/GI specimens)", "Clears debris; fungal hyphae/spores remain visible."],
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// ═════════════════════════════════════════════
// SLIDE 5 – RESPIRATORY TRACT SPECIMEN TYPES
// ═════════════════════════════════════════════
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    ["Induced Sputum", "Inhale hypertonic saline (3-5%) via nebulizer. Used when patient cannot produce sputum (TB, PCP).", "Sterile container. Label 'induced.'"],
    ["Nasopharyngeal Swab", "Flexible minitip flocked swab to posterior nasopharynx; rotate 5-10 sec. For viruses, Bordetella, Mycoplasma.", "Viral transport media (VTM/UTM). Ice for viral culture."],
    ["Nasopharyngeal Aspirate", "Suction catheter to nasopharynx. More sensitive than swabs in children.", "Mucus trap. On ice."],
    ["Bronchoalveolar Lavage (BAL)", "Bronchoscopy: 60-120 mL sterile saline instilled/aspirated from distal airways. Gold standard for PCP/fungi/TB.", "Sputum trap → sterile container. RT <2 hrs."],
    ["Bronchial Wash / Brush", "Bronchoscopic washings or protected specimen brush.", "Brush in sterile container + 1 mL saline. Fluid in trap."],
    ["Endotracheal Aspirate", "Suction catheter via endotracheal tube.", "Sputum trap/sterile container. RT <2 hrs."],
    ["Pleural Fluid", "Thoracentesis. Submit >10 mL.", "Sterile container or blood culture bottle."],
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// ═════════════════════════════════════════════
// SLIDE 6 – RESPIRATORY STAINING
// ═════════════════════════════════════════════
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    ["Gram Stain", "Bacteria (pneumococci, Klebsiella, Staph, H. influenzae), sputum quality", "Acceptable: <10 squamous cells + >25 PMNs/LPF (Bartlett). Gram+ = purple, Gram- = pink."],
    ["Ziehl-Neelsen (ZN) – Hot", "Mycobacterium tuberculosis, NTM — sputum, BAL, gastric washings", "AFB: pink-red rods on blue background. Heated carbol fuchsin."],
    ["Auramine-Phenol (Fluorochrome)", "Mycobacteria screening — higher sensitivity than ZN", "Yellow-green fluorescence. Positive screens confirmed by ZN."],
    ["Kinyoun (Cold Acid-Fast)", "Mycobacteria, Nocardia (partially AFB)", "Cold method — no heating. Same principle as ZN."],
    ["KOH Preparation", "Fungi: Aspergillus, Mucor, Candida, Cryptococcus, PCP", "Clears mucus/debris; reveals hyphae and spores."],
    ["GMS (Gomori Methenamine Silver)", "Pneumocystis jirovecii (PCP), fungi — BAL/induced sputum", "Cysts: black/dark brown on green background. Gold standard PCP."],
    ["Giemsa / Wright-Giemsa", "PCP trophozoites, Histoplasma in macrophages, viral inclusions", "Trophic forms: purple. Intracellular yeast (Histoplasma) in macrophages."],
    ["DIF (Direct Immunofluorescence)", "Legionella, influenza A/B, RSV, parainfluenza, adenovirus", "Antibody-conjugated fluorescent dye. Rapid antigen detection."],
    ["Papanicolaou (Pap)", "BAL/bronchial wash cytology — malignancy, CMV/HSV inclusions", "Nuclear and cytoplasmic detail for viral cytopathic effect."],
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// ═════════════════════════════════════════════
// SLIDE 7 – URINARY TRACT SPECIMEN TYPES
// ═════════════════════════════════════════════
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  const rows = [
    ["Mid-Stream Clean Catch (MSCC)", "Cleanse meatus. Discard first stream. Collect mid-stream 15-30 mL in sterile container.", "Sterile screw-cap. Refrigerate 2-8°C; process within 2 hrs (24 hrs if refrigerated)."],
    ["Catheter Specimen (CSU)", "Disinfect catheter port with alcohol. Aspirate with syringe from sampling port. NOT from drainage bag.", "Sterile container. Process within 2 hrs."],
    ["Suprapubic Aspirate (SPA)", "Gold standard (esp. infants). Needle inserted suprapubically under aseptic conditions.", "Sterile syringe. Immediate processing."],
    ["Urethral Swab", "Collect ≥1 hr after voiding. Insert small swab 2-4 cm into urethra, rotate, hold 2 sec.", "Specific PCR transport kits for GC/Chlamydia."],
    ["Early Morning Urine", "First void of morning — concentrated. Best for microscopy and AFB (renal TB).", "Same as MSCC. For AFB: 3 specimens on consecutive days."],
    ["3-Glass Test", "Initial stream, mid-stream, post-prostatic massage. Each centrifuged separately.", "3 sterile containers. Examine sediment with/without staining."],
    ["24-Hour Urine", "All urine over 24 hrs for quantitative studies. Schistosoma haematobium: collect noon-3 PM.", "Large sterile container ± preservative."],
  ];
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// ═════════════════════════════════════════════
// SLIDE 8 – URINARY STAINING
// ═════════════════════════════════════════════
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    ["Gram Stain (uncentrifuged urine)", "Rapid presumptive bacteriuria. 1 organism/OIF ≈ 100,000 CFU/mL.", "Gram+ cocci (Staph, Enterococcus) = purple. Gram- rods (E. coli, Klebsiella) = pink."],
    ["Gram Stain (urethral discharge)", "Gonorrhea: intracellular Gram-negative diplococci in PMNs", "Sensitivity ~95% in symptomatic males; <50% in females."],
    ["Methylene Blue / Loeffler's", "Rapid detection of bacteria and pus cells in urine", "Simple; nuclei stain dark blue, cytoplasm lighter."],
    ["Ziehl-Neelsen (AFB)", "Renal tuberculosis — urine AFB smear and culture", "Early morning urine × 3 consecutive days. AFB: pink-red on blue."],
    ["Wet Mount + Microscopy", "Trichomonas vaginalis (motile), Candida, casts, RBCs, WBCs", "Examine within minutes — motility lost quickly."],
    ["KOH Preparation", "Candida, other fungi in urine", "Clears background; yeast cells and pseudohyphae visible."],
    ["Papanicolaou (Pap)", "Urothelial cytology: TCC, viral inclusions (CMV decoy cells, BK polyomavirus)", "Voided urine cytology or bladder washings."],
    ["Modified Trichrome Blue", "Microsporidia in urine (disseminated E. intestinalis)", "Spores stain pink-reddish; also positive in GI/sputum."],
    ["Dipstick (semi-quantitative)", "Nitrite (bacteria), leukocyte esterase (pyuria), pH, protein, glucose, Hb", "Nitrite: only for nitrate-reducers (Enterobacteriaceae); 40-60% alone. Urine in bladder >4 hrs needed."],
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// ═════════════════════════════════════════════
// SLIDE 9 – MASTER STAIN QUICK REFERENCE
// ═════════════════════════════════════════════
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  headerBand(s, "Master Stain Quick Reference  —  All Three Tracts");

  const headers = ["Stain", "GI Tract", "Respiratory", "Urinary"];
  const rows = [
    ["Gram Stain",               "Bacteria, Candida",              "Bacteria, sputum quality",         "Bacteriuria, GC diplococci"],
    ["Ziehl-Neelsen (hot)",       "M. tuberculosis (gastric wash)", "M. tuberculosis, NTM",              "Renal TB (urine AFB)"],
    ["Modified ZN (cold)",        "Cryptosporidium, Cyclospora, Isospora", "Cryptosporidium in BAL",    "—"],
    ["Auramine-Phenol",           "Mycobacteria",                   "Mycobacteria (screening)",          "Mycobacteria (rare)"],
    ["Trichrome (Wheatley)",      "Entamoeba, Giardia, Blastocystis","—",                               "—"],
    ["Modified Trichrome Blue",   "Microsporidia (intestinal)",     "Microsporidia (pulmonary)",         "Microsporidia (renal)"],
    ["KOH",                       "Candida (esophageal)",           "Aspergillus, PCP, fungi",           "Candida"],
    ["GMS",                       "Intestinal fungi",               "PCP, Aspergillus",                  "—"],
    ["Calcofluor White",          "Microsporidia, Cryptosporidium", "Fungi, PCP",                       "—"],
    ["Giemsa / Wright",           "—",                              "PCP trophozoites, Histoplasma",    "Eosinophils, cytology"],
    ["India Ink",                 "—",                              "Cryptococcus (CSF/BAL)",            "—"],
    ["Wet Mount",                 "Protozoa, helminth eggs",        "—",                                 "Trichomonas, casts, cells"],
    ["Papanicolaou (Pap)",        "—",                              "Cytology, viral inclusions",        "Urothelial cytology, BK/CMV"],
  ];
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}

// ═════════════════════════════════════════════
// SLIDE 10 – MOLECULAR / ADVANCED METHODS
// ═════════════════════════════════════════════
{
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  headerBand(s, "Molecular & Advanced Diagnostic Methods");
  accentBar(s, C.teal);

  const cols = [
    {
      title: "GI TRACT",
      color: "1A7A43",
      items: [
        "FilmArray GI Panel (PCR): 22 pathogens — Salmonella, Campylobacter, Shigella, E. coli O157, Cryptosporidium, Giardia, Norovirus, Rotavirus, etc.",
        "Antigen EIA: Rotavirus, Adenovirus 40/41 (C. diff toxin EIA)",
        "NAAT for Entamoeba histolytica, Microsporidia",
        "H. pylori: rapid urease test + culture from gastric biopsy",
        "Rectal swab: acceptable for molecular enteric panel",
      ]
    },
    {
      title: "RESPIRATORY",
      color: "C05000",
      items: [
        "Respiratory Viral Panel (PCR/NAAT): Influenza A/B, RSV, SARS-CoV-2, Parainfluenza, Adenovirus, Metapneumovirus",
        "Bordetella pertussis: NP swab in specific transport",
        "Legionella urinary antigen test",
        "Pneumococcal urinary antigen test",
        "Beta-D-glucan (1,3-β-D-glucan) for invasive fungal infection in BAL",
        "Galactomannan for Aspergillus in BAL/serum",
      ]
    },
    {
      title: "URINARY TRACT",
      color: "6A1F9A",
      items: [
        "NAAT for Chlamydia trachomatis, Neisseria gonorrhoeae (urine or urethral swab)",
        "Urine culture: standard quantitative culture ≥10^5 CFU/mL = significant bacteriuria",
        "Urine cytology (Pap): for bladder carcinoma surveillance",
        "Multiplex PCR panels for UTI pathogens",
        "Urine BK virus PCR: for BK nephropathy post-transplant",
      ]
    }
  ];

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      fontSize: 10, bold: true, color: C.white, align: "center", valign: "middle", margin: 0
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      fontFace: "Calibri", valign: "top"
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}

// ═════════════════════════════════════════════
// SLIDE 11 – CONCLUSION / KEY TAKEAWAYS
// ═════════════════════════════════════════════
{
  const s = pres.addSlide();
  bg(s, C.navy);

  s.addShape(pres.shapes.RECTANGLE, {
    x: 0, y: 4.5, w: "100%", h: 1.125,
    fill: { color: C.teal }, line: { color: C.teal, width: 0 }
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    x: 0.5, y: 0.2, w: 9, h: 0.7,
    fontSize: 28, bold: true, color: C.teal, align: "center", charSpacing: 5
  });

  const takeaways = [
    { icon: "1", text: "Proper specimen collection is the single most important determinant of diagnostic result quality" },
    { icon: "2", text: "Always collect BEFORE antibiotics; label correctly; transport within recommended time windows" },
    { icon: "3", text: "Gram stain is the first-line universal stain across all three tracts; results guide empirical therapy" },
    { icon: "4", text: "Acid-fast staining (ZN / Auramine) essential for Mycobacterium in sputum, gastric wash, and urine" },
    { icon: "5", text: "Trichrome and modified ZN detect intestinal parasites; GMS and Calcofluor White for fungi/PCP" },
    { icon: "6", text: "Molecular platforms (PCR panels, NAAT) now complement and often supersede traditional staining for fastidious pathogens" },
  ];

  takeaways.forEach((t, i) => {
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    const col = i % 2;
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    const y = 1.05 + row * 1.12;
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  s.addText("Clinical Microbiology  |  Laboratory Diagnostics  |  Specimen Quality = Result Quality", {
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    fontSize: 11, color: C.navy, align: "center", italic: true, bold: true, valign: "middle"
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}

// ─────────────────────────────────────────────
// WRITE FILE
// ─────────────────────────────────────────────
pres.writeFile({ fileName: "/home/daytona/workspace/specimen-ppt/Specimen_Collection_and_Staining.pptx" })
  .then(() => console.log("✅ PPT created successfully."))
  .catch(err => console.error("❌ Error:", err));

cd /home/daytona/workspace/specimen-ppt && node create_ppt.js

node -e "const p = require('pptxgenjs'); const pres = new p(); console.log(Objec…

node -e "const p = require('pptxgenjs'); const pres = new p(); const keys = Obje…

node -e "const p = require('pptxgenjs'); const pres = new p(); const keys = Obje…

~/specimen-ppt/create_ppt.js

Successfully replaced 1 instance(s) of the string in '/home/daytona/workspace/specimen-ppt/create_ppt.js'

"ELLIPSE" in ~/specimen-ppt

"ELLIPSE" in ~/specimen-ppt

~/specimen-ppt/create_ppt.js

Successfully replaced 1 instance(s) of the string in '/home/daytona/workspace/specimen-ppt/create_ppt.js'

cd /home/daytona/workspace/specimen-ppt && node create_ppt.js

ls -lh /home/daytona/workspace/specimen-ppt/

smear preparation staining procedure steps respiratory sputum gastrointestinal stool urinary urine microbiology laboratory

https://www.cdc.gov/dpdx/diagnosticprocedures/stool/staining…

https://www.ncbi.nlm.nih.gov/books/NBK563195

Gram stain procedure steps sputum smear preparation heat fixation crystal violet iodine decolorization safranin

~/smear-ppt/create_smear_ppt.js

const pptxgen = require("pptxgenjs");
const pres = new pptxgen();
pres.layout = "LAYOUT_16x9";
pres.title = "Smear Preparation and Staining Procedures";

// ── PALETTE ──────────────────────────────────────────
const C = {
  navy:    "0B1F3A",
  teal:    "0E8A8A",
  white:   "FFFFFF",
  off:     "F2F6FA",
  silver:  "C8D6E5",
  text:    "1A1A2E",
  gi:      "1A7A43",   // green  – GI
  resp:    "B94800",   // burnt orange – Respiratory
  urine:   "5B1A8A",   // purple – Urinary
  yellow:  "F4C430",
  step:    "1565C0",   // step-number blue
};

// ── HELPERS ───────────────────────────────────────────
function bg(s, color){ s.addShape(pres.shapes.RECTANGLE,{ x:0,y:0,w:"100%",h:"100%", fill:{color}, line:{color,width:0} }); }

function headerBand(s, text, color){
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}

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    shadow:{type:"outer",color:"000000",blur:5,offset:2,angle:135,opacity:0.1} });
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  s.addTable(data,{ x,y,w,colW,rowH:0.35, border:{type:"solid",color:C.silver,pt:0.5} });
}

// ════════════════════════════════════════════════════
// SLIDE 1  – TITLE
// ════════════════════════════════════════════════════
{
  const s=pres.addSlide();
  bg(s,C.navy);
  // decorative bars
  s.addShape(pres.shapes.RECTANGLE,{x:0,y:3.6,w:"100%",h:0.07,fill:{color:C.teal},line:{color:C.teal,width:0}});
  s.addShape(pres.shapes.RECTANGLE,{x:0,y:3.68,w:"100%",h:0.04,fill:{color:C.yellow},line:{color:C.yellow,width:0}});

  s.addText("SMEAR PREPARATION",{x:0.5,y:0.6,w:9,h:0.9,fontSize:40,bold:true,color:C.white,fontFace:"Calibri",align:"center",charSpacing:3});
  s.addText("& STAINING PROCEDURES",{x:0.5,y:1.5,w:9,h:0.8,fontSize:34,bold:true,color:C.teal,fontFace:"Calibri",align:"center",charSpacing:2});

  const pills=[
    {label:"GASTROINTESTINAL",color:C.gi, x:0.55},
    {label:"RESPIRATORY",     color:C.resp,x:3.65},
    {label:"URINARY",         color:C.urine,x:6.75},
  ];
  pills.forEach(p=>{
    s.addShape(pres.shapes.ROUNDED_RECTANGLE,{x:p.x,y:2.65,w:2.7,h:0.62,fill:{color:p.color},line:{color:p.color,width:0},rectRadius:0.31});
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  });

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}

// ════════════════════════════════════════════════════
// SLIDE 2  – WHAT IS A SMEAR? + General principles
// ════════════════════════════════════════════════════
{
  const s=pres.addSlide();
  bg(s,C.off);
  headerBand(s,"What is a Smear? — General Principles",C.navy);

  // Left column – definition card
  s.addShape(pres.shapes.RECTANGLE,{x:0.28,y:1.55,w:4.5,h:3.7,fill:{color:C.white},line:{color:C.silver,width:0.75},
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  s.addText("DEFINITION",[],{x:0.28,y:1.55,w:4.5,h:0.36});
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  s.addText(
    "A smear (film) is a thin, even layer of specimen spread on a glass slide, fixed to preserve morphology, and stained so organisms and cells are visible under the microscope.\n\nSmear QUALITY is the single most important technical factor:\n• Too thick → organisms overlap; missed\n• Too thin → low yield\n• Over-fixed → poor staining uptake\n• Under-fixed → material washes off",
    {x:0.38,y:1.95,w:4.3,h:3.2,fontSize:10,color:C.text,fontFace:"Calibri",valign:"top"}
  );

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    {n:"1",t:"COLLECT — Choose the most purulent/diagnostic portion of the specimen (avoid saliva in sputum, watery stool, first stream urine)"},
    {n:"2",t:"SPREAD — Thin, even smear on a clean, grease-free, labeled glass slide"},
    {n:"3",t:"FIX — Air-dry then heat-fix (flame 2-3×) OR methanol/chemical fixation depending on stain required"},
    {n:"4",t:"STAIN — Apply staining reagents in sequence; timing is critical"},
    {n:"5",t:"RINSE — Each step requires a water rinse to remove excess reagent"},
    {n:"6",t:"DRY — Blot dry with bibulous paper; never rub"},
    {n:"7",t:"EXAMINE — Correct objective: 10x (screen) → 40x (detail) → 100x oil (definitive ID)"},
  ];
  steps.forEach((st,i)=>{
    stepBox(s,st.n,st.t,4.95,1.55+i*0.53,4.78,0.47,C.teal);
  });
}

// ════════════════════════════════════════════════════
// SLIDE 3  – GI TRACT: Direct Wet Mount
// ════════════════════════════════════════════════════
{
  const s=pres.addSlide();
  bg(s,C.off);
  headerBand(s,"Gastrointestinal Tract — Direct Wet Mount",C.gi);
  sectionTag(s,"🔬 Detects: Motile protozoa, helminth eggs/larvae",C.gi);

  const steps=[
    "Place a drop of 0.85% normal saline on a clean glass slide",
    "Transfer a match-head size of fresh stool using an applicator stick; mix to a thin suspension",
    "Apply a coverslip (22×22 mm) gently without trapping air bubbles",
    "Examine IMMEDIATELY under 10x systematically across the entire coverslip",
    "Switch to 40x for suspicious objects; watch motile objects for ≥15 sec (slow-moving amebae)",
    "Prepare a SECOND slide: replace saline with 1:5 diluted Lugol's iodine (enhances nuclear detail in cysts)",
    "Do NOT use undiluted Lugol — causes clumping; also kills motility",
  ];
  steps.forEach((st,i)=>{
    stepBox(s,i+1,st,0.28,1.55+i*0.53,9.45,0.47,C.gi);
  });

  s.addShape(pres.shapes.RECTANGLE,{x:0.28,y:5.2,w:9.45,h:0.28,fill:{color:C.gi+"22"},line:{color:C.gi,width:0.5}});
  s.addText("⚠  Examine within 30 minutes of passage. Saline = motility preserved. Iodine = nuclear detail but NO motility.",
    {x:0.38,y:5.2,w:9.25,h:0.28,fontSize:9,color:C.gi,bold:true,valign:"middle"});
}

// ════════════════════════════════════════════════════
// SLIDE 4  – GI: Concentration + Trichrome
// ════════════════════════════════════════════════════
{
  const s=pres.addSlide();
  bg(s,C.off);
  headerBand(s,"GI Tract — Concentration Technique & Trichrome Stain",C.gi);

  // LEFT – concentration
  s.addShape(pres.shapes.RECTANGLE,{x:0.28,y:1.12,w:4.6,h:0.34,fill:{color:C.gi},line:{color:C.gi,width:0}});
  s.addText("FORMALIN-ETHYL ACETATE CONCENTRATION",{x:0.28,y:1.12,w:4.6,h:0.34,fontSize:9,bold:true,color:C.white,align:"center",valign:"middle",margin:0});
  const concSteps=[
    "Emulsify ~1 g stool in 10 mL 10% formalin; strain through gauze",
    "Add 3 mL ethyl acetate; cap and shake vigorously 30 sec",
    "Centrifuge 500×g for 10 min — 4 layers form",
    "Break debris ring with stick; pour off top 3 layers",
    "Use bottom SEDIMENT for wet mount or permanent staining",
  ];
  concSteps.forEach((st,i)=>{ stepBox(s,i+1,st,0.28,1.48+i*0.62,4.6,0.56,C.gi); });

  // RIGHT – Trichrome
  s.addShape(pres.shapes.RECTANGLE,{x:5.08,y:1.12,w:4.65,h:0.34,fill:{color:C.gi},line:{color:C.gi,width:0}});
  s.addText("WHEATLEY TRICHROME STAIN (Permanent)",{x:5.08,y:1.12,w:4.65,h:0.34,fontSize:9,bold:true,color:C.white,align:"center",valign:"middle",margin:0});
  const triSteps=[
    "Smear stool (fresh/PVA-fixed) on slide; air-dry",
    "Fix: 70% ethanol + iodine → 10 min",
    "70% ethanol × 2 (5 min, then 3 min)",
    "Trichrome stain → 10 min",
    "Destain: 90% ethanol + acetic acid → 1-3 sec ONLY",
    "100% ethanol (×2, 3 min each) → Xylene (×2, 10 min)",
    "Mount coverslip; examine 100x oil — 200-300 fields",
  ];
  triSteps.forEach((st,i)=>{ stepBox(s,i+1,st,5.08,1.48+i*0.545,4.65,0.5,C.gi); });

  // Results bar
  s.addShape(pres.shapes.RECTANGLE,{x:0.28,y:5.2,w:9.45,h:0.3,fill:{color:"E8F5EE"},line:{color:C.gi,width:0.5}});
  s.addText("RESULTS — Background: green | Protozoa cytoplasm: blue-green/purple | Nuclei & chromatoids: red/purple-red | Yeast: pink-purple",
    {x:0.38,y:5.2,w:9.25,h:0.3,fontSize:8.5,color:C.gi,bold:true,valign:"middle"});
}

// ════════════════════════════════════════════════════
// SLIDE 5  – GI: Modified Kinyoun + Chromotrope
// ════════════════════════════════════════════════════
{
  const s=pres.addSlide();
  bg(s,C.off);
  headerBand(s,"GI Tract — Acid-Fast & Chromotrope Stains",C.gi);

  // LEFT – Modified Kinyoun
  s.addShape(pres.shapes.RECTANGLE,{x:0.28,y:1.12,w:4.6,h:0.36,fill:{color:C.gi},line:{color:C.gi,width:0}});
  s.addText("MODIFIED KINYOUN (Cold Acid-Fast)\nCryptosporidium | Cyclospora | Isospora",{x:0.28,y:1.12,w:4.6,h:0.36,fontSize:8.5,bold:true,color:C.white,align:"center",valign:"middle",margin:0});
  const kinySteps=[
    "Place 1-2 drops concentrated stool on slide; dry at 60°C (NOT too thick)",
    "Fix with absolute methanol — 30 seconds",
    "Flood with Kinyoun's carbol fuchsin — 1 min; brief water rinse",
    "Destain with acid-alcohol (1% HCl in 95% ethanol) — 2 min; water rinse",
    "Counterstain with malachite green (or methylene blue) — 2 min; water rinse",
    "Dry at 60°C (~5 min); mount with coverslip",
    "Examine 40x or 100x oil; scan 200-300 fields",
  ];
  kinySteps.forEach((st,i)=>{ stepBox(s,i+1,st,0.28,1.52+i*0.54,4.6,0.48,C.gi); });
  s.addShape(pres.shapes.RECTANGLE,{x:0.28,y:5.25,w:4.6,h:0.24,fill:{color:"E8F5EE"},line:{color:C.gi,width:0.5}});
  s.addText("Results: Oocysts = pink-red | Background = blue-green",{x:0.33,y:5.25,w:4.5,h:0.24,fontSize:8.5,color:C.gi,bold:true,valign:"middle"});

  // RIGHT – Chromotrope
  s.addShape(pres.shapes.RECTANGLE,{x:5.08,y:1.12,w:4.65,h:0.36,fill:{color:C.gi},line:{color:C.gi,width:0}});
  s.addText("CHROMOTROPE STAIN (Modified Trichrome)\nMicrosporidia (E. bieneusi, E. intestinalis)",{x:5.08,y:1.12,w:4.65,h:0.36,fontSize:8.5,bold:true,color:C.white,align:"center",valign:"middle",margin:0});
  const chromSteps=[
    "Place ~10 µL of 10% formalin-fixed stool on slide (unconcentrated)",
    "Heat-fix at 60°C until completely dry (5-10 min)",
    "Fix in absolute methanol — 30 seconds",
    "Perform Gram stain steps (crystal violet → iodine → decolorize) — OMIT safranin",
    "Apply chromotrope 2R working stain; heat to 50°C — 10 min",
    "Destain in acid ethanol — 10 seconds; rinse 95% then 100% ethanol",
    "Clear in xylene; mount coverslip; examine 100x oil",
  ];
  chromSteps.forEach((st,i)=>{ stepBox(s,i+1,st,5.08,1.52+i*0.54,4.65,0.48,C.gi); });
  s.addShape(pres.shapes.RECTANGLE,{x:5.08,y:5.25,w:4.65,h:0.24,fill:{color:"E8F5EE"},line:{color:C.gi,width:0.5}});
  s.addText("Results: Spores = bright pink-reddish | Background = green",{x:5.13,y:5.25,w:4.55,h:0.24,fontSize:8.5,color:C.gi,bold:true,valign:"middle"});
}

// ════════════════════════════════════════════════════
// SLIDE 6  – RESPIRATORY: Smear Preparation
// ════════════════════════════════════════════════════
{
  const s=pres.addSlide();
  bg(s,C.off);
  headerBand(s,"Respiratory Tract — Sputum Smear Preparation",C.resp);
  sectionTag(s,"Specimen: Sputum / BAL / Bronchial wash",C.resp);

  const steps=[
    "Macroscopic selection: choose the MOST PURULENT (green/yellow) portion of sputum — avoid watery, frothy or clear saliva",
    "Using a sterile wire loop or wooden applicator, transfer a small amount to the CENTER of a clean, grease-free, labeled glass slide",
    "Spread THINLY and EVENLY over an area ~2×3 cm — one smooth stroke; specimen should be barely opaque",
    "Air-dry completely at room temperature (5-10 minutes) — do NOT heat before fully dried",
    "Heat-fix: pass slide face-up THROUGH a Bunsen burner flame 2-3 times (1 second each) OR on a hot plate at 60-70°C for 5-10 min",
    "For BAL / Bronchial wash: use a cytocentrifuge (cytospin) to make a monolayer preparation for optimal cell morphology",
    "Quality check after Gram staining: ACCEPTABLE = <10 squamous epithelial cells + >25 PMNs per low-power field (10x) [Bartlett criteria]",
  ];
  steps.forEach((st,i)=>{ stepBox(s,i+1,st,0.28,1.55+i*0.53,9.45,0.47,C.resp); });
}

// ════════════════════════════════════════════════════
// SLIDE 7  – RESPIRATORY: Gram Stain
// ════════════════════════════════════════════════════
{
  const s=pres.addSlide();
  bg(s,C.off);
  headerBand(s,"Respiratory Tract — Gram Stain Procedure",C.resp);
  sectionTag(s,"First-line universal stain for all specimens",C.resp);

  // Steps (left 2/3)
  const steps=[
    {n:"1",t:"Flood air-dried, heat-fixed smear with CRYSTAL VIOLET (primary stain) — 1 minute"},
    {n:"2",t:"Wash: gentle indirect stream of tap water — 2 seconds"},
    {n:"3",t:"Flood with GRAM'S IODINE (mordant — fixes crystal violet-iodine complex in cell wall) — 1 minute"},
    {n:"4",t:"Wash: gentle water rinse — 2 seconds"},
    {n:"5",t:"DECOLORIZE with 95% ethanol/acetone — 15-30 sec or drop by drop until runoff runs CLEAR (CRITICAL — do not over-decolorize)"},
    {n:"6",t:"Wash IMMEDIATELY with water to stop decolorization"},
    {n:"7",t:"Flood with SAFRANIN (counterstain) — 30 seconds to 1 minute"},
    {n:"8",t:"Wash gently until no color in runoff; BLOT DRY (do not rub); examine under 100x oil immersion"},
  ];
  steps.forEach((st,i)=>{ stepBox(s,st.n,st.t,0.28,1.12+i*0.53,6.4,0.47,C.resp); });

  // Results panel (right)
  s.addShape(pres.shapes.RECTANGLE,{x:6.85,y:1.12,w:2.9,h:0.36,fill:{color:C.resp},line:{color:C.resp,width:0}});
  s.addText("RESULTS",{x:6.85,y:1.12,w:2.9,h:0.36,fontSize:10,bold:true,color:C.white,align:"center",valign:"middle",margin:0});
  const res=[
    ["Gram + (PURPLE)","Strep, Staph, Pneumococcus"],
    ["Gram − (PINK)","Klebsiella, E. coli, H. flu"],
    ["Diplococci","Pneumococcus (lancet-shaped)"],
    ["Curved rods","Campylobacter"],
    ["Yeast (oval)","Candida spp."],
    ["Squamous cells","Saliva — reject specimen"],
  ];
  res.forEach((r,i)=>{
    const fy=1.5+i*0.52;
    s.addShape(pres.shapes.RECTANGLE,{x:6.85,y:fy,w:2.9,h:0.5,fill:{color:i%2===0?C.off:C.white},line:{color:C.silver,width:0.5}});
    s.addText(r[0],{x:6.92,y:fy+0.02,w:2.75,h:0.22,fontSize:9,bold:true,color:C.resp});
    s.addText(r[1],{x:6.92,y:fy+0.24,w:2.75,h:0.2,fontSize:8.5,color:C.text});
  });
}

// ════════════════════════════════════════════════════
// SLIDE 8  – RESPIRATORY: Ziehl-Neelsen
// ════════════════════════════════════════════════════
{
  const s=pres.addSlide();
  bg(s,C.off);
  headerBand(s,"Respiratory Tract — Ziehl-Neelsen (AFB) Stain",C.resp);
  sectionTag(s,"Mycobacterium tuberculosis / NTM",C.resp);

  const steps=[
    {n:"1",t:"Prepare smear as usual; allow to dry THOROUGHLY (incomplete drying = poor AFB staining); heat-fix"},
    {n:"2",t:"Flood smear with CARBOL FUCHSIN (basic fuchsin + phenol solution)"},
    {n:"3",t:"HEAT slide by passing flame underneath until STEAM RISES — maintain steaming for 5 minutes; reapply stain as it dries (do NOT boil)"},
    {n:"4",t:"COOL slide for 5 minutes; wash gently with distilled water"},
    {n:"5",t:"DECOLORIZE with ACID-ALCOHOL (3% HCl in 95% ethanol) until smear appears pale pink — approximately 2-3 minutes; wash with water"},
    {n:"6",t:"COUNTERSTAIN with methylene blue (or malachite green) — 1-2 minutes; wash with water"},
    {n:"7",t:"Blot dry; air-dry; examine 100x oil immersion — scan 100-300 fields (300 fields = 1 cm²)"},
  ];
  steps.forEach((st,i)=>{ stepBox(s,st.n,st.t,0.28,1.12+i*0.56,9.45,0.5,C.resp); });

  // Results + Grading bar
  s.addShape(pres.shapes.RECTANGLE,{x:0.28,y:5.1,w:9.45,h:0.38,fill:{color:"FFF3E0"},line:{color:C.resp,width:0.75}});
  s.addText(
    "RESULTS: AFB = bright PINK-RED rods | Background = BLUE (methylene blue)\n"+
    "GRADING (WHO): Scanty 1-9/100 fields | 1+ = 10-99/100 fields | 2+ = 1-10/field | 3+ = >10/field",
    {x:0.38,y:5.1,w:9.25,h:0.38,fontSize:9,color:C.resp,bold:true,valign:"middle"}
  );
}

// ════════════════════════════════════════════════════
// SLIDE 9  – RESPIRATORY: Auramine + GMS
// ════════════════════════════════════════════════════
{
  const s=pres.addSlide();
  bg(s,C.off);
  headerBand(s,"Respiratory Tract — Auramine-Phenol & GMS Stains",C.resp);

  // LEFT – Auramine
  s.addShape(pres.shapes.RECTANGLE,{x:0.28,y:1.12,w:4.6,h:0.36,fill:{color:C.resp},line:{color:C.resp,width:0}});
  s.addText("AURAMINE-PHENOL (Fluorochrome)\nMore sensitive than ZN for AFB screening",{x:0.28,y:1.12,w:4.6,h:0.36,fontSize:8.5,bold:true,color:C.white,align:"center",valign:"middle",margin:0});
  const aurSteps=[
    "Prepare and heat-fix smear as usual",
    "Flood with auramine-phenol stain — 15 minutes; wash with water",
    "Decolorize with acid-alcohol — 2-3 minutes; wash with water",
    "Counterstain with 0.5% potassium permanganate — 2 min (quenches background fluorescence); wash; blot dry",
    "Examine under FLUORESCENCE MICROSCOPE (blue-violet excitation, 450-490 nm)",
    "AFB: bright yellow-green / orange on DARK background",
    "All positive auramine screens must be CONFIRMED by ZN",
  ];
  aurSteps.forEach((st,i)=>{ stepBox(s,i+1,st,0.28,1.52+i*0.54,4.6,0.48,C.resp); });

  // RIGHT – GMS
  s.addShape(pres.shapes.RECTANGLE,{x:5.08,y:1.12,w:4.65,h:0.36,fill:{color:C.resp},line:{color:C.resp,width:0}});
  s.addText("GOMORI METHENAMINE SILVER (GMS)\nPneumocystis jirovecii (PCP) / Fungi in BAL",{x:5.08,y:1.12,w:4.65,h:0.36,fontSize:8.5,bold:true,color:C.white,align:"center",valign:"middle",margin:0});
  const gmsSteps=[
    "Prepare BAL smear via cytocentrifuge (cytospin monolayer)",
    "Fix in 95% ethanol",
    "Oxidize with periodic acid or chromic acid (opens cell wall ring structures)",
    "Treat with methenamine silver nitrate solution at 60°C for 30-60 min",
    "Rinse; tone with gold chloride (optional)",
    "Counterstain with light green",
    "Results: Fungal walls + PCP cysts = BLACK/dark brown | Background = green",
  ];
  gmsSteps.forEach((st,i)=>{ stepBox(s,i+1,st,5.08,1.52+i*0.54,4.65,0.48,C.resp); });
}

// ════════════════════════════════════════════════════
// SLIDE 10 – URINARY TRACT: Smear Preparation
// ════════════════════════════════════════════════════
{
  const s=pres.addSlide();
  bg(s,C.off);
  headerBand(s,"Urinary Tract — Urine Smear Preparation",C.urine);
  sectionTag(s,"Specimen: Mid-stream clean catch / Catheter / SPA",C.urine);

  // Two methods side by side
  s.addShape(pres.shapes.RECTANGLE,{x:0.28,y:1.12,w:4.6,h:0.34,fill:{color:C.urine},line:{color:C.urine,width:0}});
  s.addText("METHOD 1 — Centrifuged Sediment",{x:0.28,y:1.12,w:4.6,h:0.34,fontSize:9,bold:true,color:C.white,align:"center",valign:"middle",margin:0});
  const centSteps=[
    "Mix urine well; transfer 10 mL into conical centrifuge tube",
    "Centrifuge at 2000 rpm (400×g) for 5 minutes",
    "Decant supernatant; leave ~0.5-1 mL",
    "Resuspend sediment by tapping / flicking tube",
    "Transfer ONE DROP to a clean glass slide",
    "For WET MOUNT: apply coverslip; examine immediately",
    "For STAINED SMEAR: spread evenly; air-dry; heat-fix or methanol-fix",
  ];
  centSteps.forEach((st,i)=>{ stepBox(s,i+1,st,0.28,1.48+i*0.54,4.6,0.48,C.urine); });

  s.addShape(pres.shapes.RECTANGLE,{x:5.08,y:1.12,w:4.65,h:0.34,fill:{color:C.urine},line:{color:C.urine,width:0}});
  s.addText("METHOD 2 — Uncentrifuged Urine (Gram Stain Screening)",{x:5.08,y:1.12,w:4.65,h:0.34,fontSize:9,bold:true,color:C.white,align:"center",valign:"middle",margin:0});
  const uncentSteps=[
    "Use well-mixed, UNCENTRIFUGED midstream urine",
    "Place ONE DROP on a clean glass slide",
    "Spread thinly and evenly (~2×3 cm area)",
    "Air-dry completely at room temperature",
    "Heat-fix: pass through flame 2-3 times OR hot plate 60°C / 5 min",
    "Proceed directly to Gram stain",
    "⚠  1 organism per oil-immersion field ≈ 100,000 CFU/mL (significant bacteriuria)",
  ];
  uncentSteps.forEach((st,i)=>{ stepBox(s,i+1,st,5.08,1.48+i*0.54,4.65,0.48,C.urine); });

  s.addShape(pres.shapes.RECTANGLE,{x:0.28,y:5.25,w:9.45,h:0.26,fill:{color:"F3EEF8"},line:{color:C.urine,width:0.5}});
  s.addText("KEY: Early morning urine = best for microscopy — examine within MINUTES (cells lyse in dilute/alkaline urine; Trichomonas motility lost rapidly)",
    {x:0.38,y:5.25,w:9.25,h:0.26,fontSize:9,color:C.urine,bold:true,valign:"middle"});
}

// ════════════════════════════════════════════════════
// SLIDE 11 – URINARY: Wet Mount + Gram Stain
// ════════════════════════════════════════════════════
{
  const s=pres.addSlide();
  bg(s,C.off);
  headerBand(s,"Urinary Tract — Wet Mount & Gram Stain",C.urine);

  // LEFT – Wet Mount
  s.addShape(pres.shapes.RECTANGLE,{x:0.28,y:1.12,w:4.6,h:0.34,fill:{color:C.urine},line:{color:C.urine,width:0}});
  s.addText("WET MOUNT OF URINE SEDIMENT (Unstained)",{x:0.28,y:1.12,w:4.6,h:0.34,fontSize:9,bold:true,color:C.white,align:"center",valign:"middle",margin:0});
  const wmSteps=[
    "Centrifuge 10 mL urine at 2000 rpm for 5 min; decant supernatant",
    "Resuspend sediment; place one drop on slide; apply coverslip",
    "Examine under 10x (low power) then 40x (high power) — IMMEDIATELY",
    "Look for: Trichomonas vaginalis (pear-shaped, jerky motility with flagella)",
    "RBCs, WBCs, casts (hyaline, granular, RBC casts = glomerulonephritis)",
    "Epithelial cells (squamous = contamination; tubular = significant)",
    "Bacteria, yeast cells (Candida — budding oval), crystals",
  ];
  wmSteps.forEach((st,i)=>{ stepBox(s,i+1,st,0.28,1.48+i*0.54,4.6,0.48,C.urine); });

  // RIGHT – Gram stain for urine
  s.addShape(pres.shapes.RECTANGLE,{x:5.08,y:1.12,w:4.65,h:0.34,fill:{color:C.urine},line:{color:C.urine,width:0}});
  s.addText("GRAM STAIN OF URINE (Steps same as Respiratory)",{x:5.08,y:1.12,w:4.65,h:0.34,fontSize:9,bold:true,color:C.white,align:"center",valign:"middle",margin:0});
  const gramUSteps=[
    "Flood heat-fixed smear with CRYSTAL VIOLET — 1 min; water wash 2 sec",
    "Flood with GRAM'S IODINE — 1 min; water wash 2 sec",
    "DECOLORIZE with 95% ethanol/acetone — 15-30 sec until runoff clear",
    "Water wash IMMEDIATELY to stop decolorization",
    "Counterstain with SAFRANIN — 30-60 sec; water rinse; blot dry",
    "Examine 100x oil immersion",
  ];
  gramUSteps.forEach((st,i)=>{ stepBox(s,i+1,st,5.08,1.48+i*0.545,4.65,0.5,C.urine); });

  // Results
  s.addShape(pres.shapes.RECTANGLE,{x:5.08,y:4.82,w:4.65,h:0.66,fill:{color:"F3EEF8"},line:{color:C.urine,width:0.5}});
  s.addText(
    "RESULTS: Gram+ cocci (purple) = Staph, Enterococcus | Gram− rods (pink) = E. coli, Klebsiella, Proteus\n"+
    "Intracellular Gram− diplococci in PMNs = Neisseria gonorrhoeae (urethral discharge)",
    {x:5.13,y:4.82,w:4.55,h:0.66,fontSize:8.5,color:C.urine,fontFace:"Calibri",valign:"middle"}
  );
}

// ════════════════════════════════════════════════════
// SLIDE 12 – URINARY: ZN + Pap stain
// ════════════════════════════════════════════════════
{
  const s=pres.addSlide();
  bg(s,C.off);
  headerBand(s,"Urinary Tract — AFB (ZN) & Papanicolaou Stain",C.urine);

  // LEFT – ZN urine
  s.addShape(pres.shapes.RECTANGLE,{x:0.28,y:1.12,w:4.6,h:0.34,fill:{color:C.urine},line:{color:C.urine,width:0}});
  s.addText("ZIEHL-NEELSEN (AFB) — Renal TB",{x:0.28,y:1.12,w:4.6,h:0.34,fontSize:9,bold:true,color:C.white,align:"center",valign:"middle",margin:0});
  const znUSteps=[
    "Collect FIRST MORNING midstream clean-catch urine × 3 consecutive days",
    "Centrifuge LARGE volume (50-100 mL) at high speed to concentrate sediment",
    "Prepare thin smear from sediment; air-dry; heat-fix",
    "Flood with carbol fuchsin; heat until steam rises — 5 minutes",
    "Cool, wash; decolorize with acid-alcohol (3% HCl) — 2-3 min; wash",
    "Counterstain with methylene blue — 1-2 min; wash; blot dry",
    "Examine 100x oil; report using WHO grading scale",
  ];
  znUSteps.forEach((st,i)=>{ stepBox(s,i+1,st,0.28,1.48+i*0.54,4.6,0.48,C.urine); });
  s.addShape(pres.shapes.RECTANGLE,{x:0.28,y:5.25,w:4.6,h:0.24,fill:{color:"F3EEF8"},line:{color:C.urine,width:0.5}});
  s.addText("Results: AFB = pink-red on blue background",{x:0.33,y:5.25,w:4.5,h:0.24,fontSize:8.5,color:C.urine,bold:true,valign:"middle"});

  // RIGHT – Pap stain
  s.addShape(pres.shapes.RECTANGLE,{x:5.08,y:1.12,w:4.65,h:0.34,fill:{color:C.urine},line:{color:C.urine,width:0}});
  s.addText("PAPANICOLAOU (PAP) STAIN — Urine Cytology",{x:5.08,y:1.12,w:4.65,h:0.34,fontSize:9,bold:true,color:C.white,align:"center",valign:"middle",margin:0});
  const papSteps=[
    "Collect voided urine or bladder washing; centrifuge to concentrate",
    "Prepare cytospin (cytocentrifuge) preparation for monolayer",
    "Fix IMMEDIATELY in 95% ethanol or spray fixative — 15 minutes",
    "Hydrate through graded alcohols to water",
    "Stain nuclei with HEMATOXYLIN (Harris/modified) — 3-5 min; wash; blue in ammonia",
    "Counterstain cytoplasm with ORANGE G-6 then EA (Eosin Azure) solutions",
    "Dehydrate; clear in xylene; mount; examine 40x and 100x",
  ];
  papSteps.forEach((st,i)=>{ stepBox(s,i+1,st,5.08,1.48+i*0.54,4.65,0.48,C.urine); });
  s.addShape(pres.shapes.RECTANGLE,{x:5.08,y:5.25,w:4.65,h:0.24,fill:{color:"F3EEF8"},line:{color:C.urine,width:0.5}});
  s.addText("Detects: TCC, CMV decoy cells, BK polyomavirus inclusions, dysplastic cells",{x:5.13,y:5.25,w:4.55,h:0.24,fontSize:8.5,color:C.urine,bold:true,valign:"middle"});
}

// ════════════════════════════════════════════════════
// SLIDE 13 – COMPARISON SUMMARY TABLE
// ════════════════════════════════════════════════════
{
  const s=pres.addSlide();
  bg(s,C.off);
  headerBand(s,"Smear Preparation & Staining — Quick Reference Comparison",C.navy);

  const headers=["Stain","Fixation","Key Steps (Sequence)","Result Colors","Tract(s)"];
  const rows=[
    ["Wet Mount","None","Saline/Iodine → coverslip → immediate exam","Organisms: translucent; iodine enhances cysts","GI, Urine"],
    ["Gram Stain","Heat-fix","Crystal violet → Iodine → Decolorize → Safranin","G+ = purple | G− = pink","ALL 3"],
    ["Ziehl-Neelsen (Hot)","Heat-fix","Carbol fuchsin (heated) → Acid-alcohol → Methylene blue","AFB = pink-red | Bg = blue","GI, Resp, Urine"],
    ["Modified Kinyoun (Cold)","Methanol-fix","Carbol fuchsin (cold) → Acid-alcohol → Malachite green","Oocysts = pink-red | Bg = blue-green","GI, Resp"],
    ["Wheatley Trichrome","PVA / Schaudinn","EtOH-iodine → Trichrome 10min → Acid-EtOH destain → Xylene","Protozoa = blue-green | Nuclei = red","GI"],
    ["Chromotrope","Methanol-fix","Gram steps (no safranin) → Chromotrope 50°C → Acid-EtOH","Spores = pink-red | Bg = green","GI, Urine"],
    ["Auramine-Phenol","Heat-fix","Auramine-phenol → Acid-alcohol → KMnO₄ counterstain","AFB = yellow-green (fluorescence)","Resp"],
    ["GMS","EtOH fix","Oxidize → Methenamine silver 60°C → Light green counterstain","Fungi/PCP = black | Bg = green","Resp"],
    ["Papanicolaou (Pap)","Immediate EtOH","Hematoxylin → Orange G → Eosin Azure → Xylene","Nuclei = blue | Cytoplasm = pink-green","Resp, Urine"],
    ["KOH Prep","None","Mix with 10-20% KOH → warm gently → coverslip","Fungal hyphae/yeast visible (clear)","ALL 3"],
  ];
  addTable(s,headers,rows,0.18,1.14,9.65,[1.45,1.3,3.2,2.0,1.7]);
}

// ════════════════════════════════════════════════════
// SLIDE 14 – CONCLUSION
// ════════════════════════════════════════════════════
{
  const s=pres.addSlide();
  bg(s,C.navy);
  s.addShape(pres.shapes.RECTANGLE,{x:0,y:4.5,w:"100%",h:1.125,fill:{color:C.teal},line:{color:C.teal,width:0}});
  s.addShape(pres.shapes.RECTANGLE,{x:0,y:3.7,w:"100%",h:0.07,fill:{color:C.yellow},line:{color:C.yellow,width:0}});

  s.addText("KEY TAKEAWAYS",{x:0.5,y:0.2,w:9,h:0.65,fontSize:26,bold:true,color:C.teal,align:"center",charSpacing:5});

  const pts=[
    {n:"1",t:"Smear QUALITY is paramount — select the right portion, spread thin, fix properly before staining"},
    {n:"2",t:"GRAM STAIN is the universal first-line stain for all three tracts; results guide empirical antibiotic therapy"},
    {n:"3",t:"GI TRACT: Wet mount (fresh; immediate) + Trichrome (protozoa) + Modified ZN (coccidia) + Chromotrope (microsporidia)"},
    {n:"4",t:"RESPIRATORY: Gram stain (quality check + bacteria) → ZN/Auramine (AFB/TB) → GMS (PCP/fungi in BAL)"},
    {n:"5",t:"URINARY: Centrifuge sediment → Wet mount (Trichomonas; immediate!) → Gram stain → ZN (renal TB, ×3 days)"},
    {n:"6",t:"DECOLORIZATION is the CRITICAL step in both Gram and ZN — over-decolorize and you lose your result"},
  ];
  pts.forEach((p,i)=>{
    const col=i%2; const row=Math.floor(i/2);
    const x=0.3+col*4.85; const y=1.0+row*1.1;
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    s.addText(p.n,{x,y,w:0.36,h:0.36,fontSize:11,bold:true,color:C.navy,align:"center",valign:"middle",margin:0});
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  });

  s.addText("Smear Preparation & Staining  |  GI · Respiratory · Urinary  |  Lab Microbiology",
    {x:0.5,y:4.6,w:9,h:0.45,fontSize:11,color:C.navy,align:"center",italic:true,bold:true,valign:"middle"});
}

// ── WRITE ─────────────────────────────────────────────
pres.writeFile({fileName:"/home/daytona/workspace/smear-ppt/Smear_Preparation_and_Staining.pptx"})
  .then(()=>console.log("✅ PPT created successfully."))
  .catch(err=>console.error("❌",err));

cd /home/daytona/workspace/smear-ppt && node create_smear_ppt.js

ls -lh /home/daytona/workspace/smear-ppt/

RGUHS 3rd BSc MLT syllabus clinical microbiology smear preparation staining respiratory GI urinary tract

RGUHS BSc MLT 3rd year Microbiology II syllabus specimen processing smear preparation staining 2023 2024

~/rguhs-ppt/create_rguhs_ppt.js

const pptxgen = require("pptxgenjs");
const pres = new pptxgen();
pres.layout = "LAYOUT_16x9";
pres.title = "Smear Preparation and Staining - 3rd BSc MLT RGUHS";

// ── PALETTE ──────────────────────────────────────────
const C = {
  navy:   "0B2545",
  teal:   "0E8A8A",
  white:  "FFFFFF",
  off:    "F0F4F8",
  silver: "C8D6E5",
  text:   "1A1A2E",
  gi:     "1A7A43",
  resp:   "B94800",
  urine:  "5B1A8A",
  yellow: "F4C430",
  red:    "C0392B",
  ltgray: "E8EEF4",
};

// ── HELPERS ──────────────────────────────────────────
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}
function colHdr(s, x, y, w, h, txt, col) {
  s.addShape(pres.shapes.RECTANGLE, { x, y, w, h, fill:{color:col}, line:{color:col,width:0} });
  s.addText(txt, { x, y, w, h, fontSize:9, bold:true, color:C.white, align:"center", valign:"middle", margin:0 });
}
function tbl(s, headers, rows, x, y, w, colW) {
  const data = [];
  data.push(headers.map(h => ({ text:h, options:{bold:true,color:C.white,fill:C.navy,fontSize:9,align:"center",valign:"middle"} })));
  rows.forEach((r,i) => data.push(r.map(cell => ({ text:cell, options:{color:C.text,fill:i%2===0?C.off:C.white,fontSize:8.5,valign:"middle"} }))));
  s.addTable(data, { x, y, w, colW, rowH:0.33, border:{type:"solid",color:C.silver,pt:0.5} });
}

// ════════════════════════════════════════════
// SLIDE 1 — TITLE
// ════════════════════════════════════════════
{
  const s = pres.addSlide();
  bg(s, C.navy);
  s.addShape(pres.shapes.RECTANGLE, {x:0, y:3.55, w:"100%", h:0.07, fill:{color:C.teal}, line:{color:C.teal,width:0}});
  s.addShape(pres.shapes.RECTANGLE, {x:0, y:3.63, w:"100%", h:0.04, fill:{color:C.yellow}, line:{color:C.yellow,width:0}});

  s.addText("SMEAR PREPARATION", { x:0.5, y:0.45, w:9, h:0.85, fontSize:38, bold:true, color:C.white, align:"center", charSpacing:3, fontFace:"Calibri" });
  s.addText("& STAINING PROCEDURES", { x:0.5, y:1.28, w:9, h:0.75, fontSize:32, bold:true, color:C.teal, align:"center", charSpacing:2, fontFace:"Calibri" });

  // University badge
  s.addShape(pres.shapes.ROUNDED_RECTANGLE, {x:2.8, y:2.2, w:4.4, h:1.1, fill:{color:"142850"}, line:{color:C.teal,width:1.5}, rectRadius:0.12});
  s.addText([
    {text:"3rd B.Sc. MLT  |  RGUHS\n", options:{bold:true, fontSize:13, color:C.yellow}},
    {text:"Microbiology II — Specimen Processing", options:{fontSize:10.5, color:C.silver}},
  ], { x:2.8, y:2.2, w:4.4, h:1.1, align:"center", valign:"middle" });

  // 3 organ pills
  const pills = [
    {label:"GASTROINTESTINAL TRACT", col:C.gi,    x:0.45},
    {label:"RESPIRATORY TRACT",      col:C.resp,  x:3.65},
    {label:"URINARY TRACT",          col:C.urine, x:6.85},
  ];
  pills.forEach(p => {
    s.addShape(pres.shapes.ROUNDED_RECTANGLE, {x:p.x, y:3.75, w:2.7, h:0.58, fill:{color:p.col}, line:{color:p.col,width:0}, rectRadius:0.29});
    s.addText(p.label, {x:p.x, y:3.75, w:2.7, h:0.58, fontSize:8.5, bold:true, color:C.white, align:"center", valign:"middle", margin:0});
  });
  s.addText("Ananthanarayan & Paniker  |  Mackie & McCartney  |  RGUHS Prescribed", {x:0.5, y:5.05, w:9, h:0.35, fontSize:9.5, color:C.silver, align:"center", italic:true});
}

// ════════════════════════════════════════════
// SLIDE 2 — UNIT OVERVIEW + KEY DEFINITIONS
// ════════════════════════════════════════════
{
  const s = pres.addSlide();
  bg(s, C.off);
  hdr(s, "Unit Overview — Specimen Processing (Microbiology II)", C.navy);

  // Left — what is a smear
  colHdr(s, 0.28, 1.08, 4.55, 0.34, "WHAT IS A SMEAR?", C.navy);
  card(s, 0.28, 1.44, 4.55, 2.4);
  s.addText(
    "A smear (film) is a thin, even layer of specimen spread on a glass slide, fixed to preserve morphology, and stained with dyes so organisms and cells become visible under the microscope.\n\n• Too THICK → organisms overlap, missed\n• Too THIN → low yield\n• Over-fixed → poor staining\n• Under-fixed → material washes off\n\nSMEAR QUALITY is the most important factor in any staining procedure.",
    {x:0.36, y:1.5, w:4.38, h:2.28, fontSize:10, color:C.text, fontFace:"Calibri", valign:"top"}
  );

  // Right — 7 universal steps
  colHdr(s, 5.0, 1.08, 4.73, 0.34, "7 UNIVERSAL STEPS", C.navy);
  const steps = [
    "SELECT the best portion of specimen",
    "SPREAD — thin, even smear on clean labeled slide",
    "AIR-DRY completely at room temperature",
    "HEAT-FIX — flame 2-3× (1 sec each) or hot plate 60°C",
    "APPLY stain reagents in sequence with correct timing",
    "RINSE with water after each step",
    "BLOT DRY — never rub; examine under correct objective",
  ];
  steps.forEach((st,i) => {
    const y = 1.44 + i*0.48;
    s.addShape(pres.shapes.RECTANGLE, {x:5.0, y, w:4.73, h:0.46, fill:{color:i%2===0?C.off:C.white}, line:{color:C.silver,width:0.5}});
    s.addShape(pres.shapes.OVAL, {x:5.06, y:y+0.08, w:0.28, h:0.28, fill:{color:C.teal}, line:{color:C.teal,width:0}});
    s.addText(String(i+1), {x:5.06, y:y+0.08, w:0.28, h:0.28, fontSize:8.5, bold:true, color:C.white, align:"center", valign:"middle", margin:0});
    s.addText(st, {x:5.4, y:y+0.06, w:4.25, h:0.34, fontSize:9.5, color:C.text, valign:"middle"});
  });
}

// ════════════════════════════════════════════
// SLIDE 3 — GI SPECIMENS + DIRECT WET MOUNT
// ════════════════════════════════════════════
{
  const s = pres.addSlide();
  bg(s, C.off);
  hdr(s, "Gastrointestinal Tract — Specimens & Direct Wet Mount", C.gi);

  // Specimen table left
  colHdr(s, 0.28, 1.08, 4.55, 0.32, "GI TRACT SPECIMENS", C.gi);
  tbl(s,
    ["Specimen","Source","Used For"],
    [
      ["Stool (feces)","Freshly passed","Bacteria, parasites, viruses"],
      ["Rectal swab","Rectum","Shigella, Vibrio, enteric bacteria"],
      ["Gastric lavage","Stomach washings","M. tuberculosis (pauci-bacillary)"],
      ["Duodenal aspirate","Upper small intestine","Giardia, Strongyloides"],
      ["Sigmoidoscopy scraping","Colonic mucosa","Entamoeba histolytica"],
    ],
    0.28, 1.42, 4.55, [1.3, 1.5, 1.75]
  );

  // Direct wet mount steps right
  colHdr(s, 5.0, 1.08, 4.73, 0.32, "DIRECT WET MOUNT (Smear Prep)", C.gi);
  const wmSteps = [
    "Place a drop of 0.85% NORMAL SALINE on clean glass slide",
    "Transfer match-head size of fresh stool using applicator stick",
    "Mix to a thin, uniform suspension; apply coverslip (22×22 mm)",
    "Examine IMMEDIATELY under 10x (screen), then 40x (detail)",
    "Watch motile amebae for ≥15 seconds (slow-moving!)",
    "2nd slide: use 1:5 DILUTED LUGOL'S IODINE instead of saline (nuclear detail)",
    "Do NOT use undiluted Lugol — causes clumping",
  ];
  wmSteps.forEach((st,i) => stepDot(s, i+1, st, 5.0, 1.42+i*0.53, 4.73, C.gi));

  // Alert bar
  s.addShape(pres.shapes.RECTANGLE, {x:5.0, y:5.16, w:4.73, h:0.3, fill:{color:"E8F5EE"}, line:{color:C.gi,width:0.75}});
  s.addText("⚠  Saline = motility preserved | Iodine = nuclear detail but kills motility | Examine within 30 min",
    {x:5.08, y:5.16, w:4.6, h:0.3, fontSize:8.5, color:C.gi, bold:true, valign:"middle"});
}

// ════════════════════════════════════════════
// SLIDE 4 — CONCENTRATION + TRICHROME
// ════════════════════════════════════════════
{
  const s = pres.addSlide();
  bg(s, C.off);
  hdr(s, "GI Tract — Concentration Technique & Trichrome Stain", C.gi);

  // Left — Concentration
  colHdr(s, 0.28, 1.05, 4.55, 0.34, "FORMALIN-ETHYL ACETATE CONCENTRATION", C.gi);
  const concSteps = [
    "Emulsify ~1g stool in 10 mL 10% formalin; strain through gauze",
    "Add 3 mL ethyl acetate; cap and shake vigorously 30 sec",
    "Centrifuge 500×g for 10 min → 4 layers form",
    "Layers: (1) Ethyl acetate plug  (2) Formalin  (3) Debris ring  (4) SEDIMENT",
    "Break debris ring; discard top 3 layers",
    "Use bottom SEDIMENT for wet mount or permanent staining",
  ];
  concSteps.forEach((st,i) => stepDot(s, i+1, st, 0.28, 1.42+i*0.56, 4.55, C.gi));
  s.addShape(pres.shapes.RECTANGLE, {x:0.28, y:4.62, w:4.55, h:0.28, fill:{color:"E8F5EE"}, line:{color:C.gi,width:0.5}});
  s.addText("Purpose: ↑ sensitivity for protozoan cysts + helminth eggs/larvae",
    {x:0.36, y:4.62, w:4.38, h:0.28, fontSize:9, color:C.gi, bold:true, valign:"middle"});

  // Right — Trichrome procedure table
  colHdr(s, 5.0, 1.05, 4.73, 0.34, "WHEATLEY TRICHROME STAIN (Permanent)", C.gi);
  tbl(s,
    ["Step","Reagent","Time"],
    [
      ["1. Fix","70% ethanol + iodine","10 min"],
      ["2. Hydrate","70% ethanol","5 min"],
      ["3. Hydrate","70% ethanol","3 min"],
      ["4. STAIN","Trichrome stain","10 min"],
      ["5. DESTAIN ⚠","90% ethanol + acetic acid","1–3 sec ONLY"],
      ["6. Dehydrate","100% ethanol (×2)","3 min each"],
      ["7. Clear","Xylene (×2)","10 min each"],
      ["8. Mount & View","Permount + coverslip","100x oil, 200-300 fields"],
    ],
    5.0, 1.42, 4.73, [1.4, 2.1, 1.23]
  );
  // Results
  s.addShape(pres.shapes.RECTANGLE, {x:5.0, y:4.56, w:4.73, h:0.9, fill:{color:"E8F5EE"}, line:{color:C.gi,width:0.75}});
  s.addText("RESULTS\n• Background → GREEN\n• Protozoa cytoplasm → BLUE-GREEN to PURPLE\n• Nuclei & chromatoid bodies → RED/PURPLE-RED\n• Yeast → PINK to PURPLE",
    {x:5.08, y:4.56, w:4.58, h:0.9, fontSize:9, color:C.gi, fontFace:"Calibri", valign:"top"});
}

// ════════════════════════════════════════════
// SLIDE 5 — MODIFIED KINYOUN
// ════════════════════════════════════════════
{
  const s = pres.addSlide();
  bg(s, C.off);
  hdr(s, "GI Tract — Modified Kinyoun (Cold Acid-Fast) Stain", C.gi);

  // Principle
  colHdr(s, 0.28, 1.05, 9.45, 0.32, "PRINCIPLE", C.gi);
  s.addText(
    "Coccidian oocysts (Cryptosporidium, Cyclospora, Cystoisospora/Isospora) have a WAXY wall resistant to decolorization with DILUTE acid-alcohol. They retain carbol fuchsin (pink-red) while the background is decolorized. NO HEAT is required — this is the COLD method.",
    {x:0.28, y:1.4, w:9.45, h:0.5, fontSize:10.5, color:C.text, fontFace:"Calibri", valign:"middle"}
  );

  // Two columns: reagents + steps
  colHdr(s, 0.28, 1.97, 2.5, 0.32, "REAGENTS", C.gi);
  const reags = ["Kinyoun's carbol fuchsin (primary stain)","Acid-alcohol: 1% HCl in 95% ethanol (decolorizer)","Malachite green or methylene blue (counterstain)","Absolute methanol (fixative)"];
  reags.forEach((r,i) => {
    s.addShape(pres.shapes.RECTANGLE, {x:0.28, y:2.31+i*0.42, w:2.5, h:0.4, fill:{color:i%2===0?C.off:C.white}, line:{color:C.silver,width:0.5}});
    s.addText(`• ${r}`, {x:0.36, y:2.31+i*0.42, w:2.34, h:0.4, fontSize:9, color:C.text, valign:"middle"});
  });

  colHdr(s, 2.92, 1.97, 6.81, 0.32, "PROCEDURE (Step-by-Step)", C.gi);
  const kSteps = [
    "Place 1-2 drops concentrated stool on slide; dry at 60°C (NOT too thick)",
    "Fix with ABSOLUTE METHANOL — 30 seconds",
    "Flood with Kinyoun's carbol fuchsin — 1 minute; brief water rinse",
    "Decolorize with ACID-ALCOHOL (1% HCl in 95% ethanol) — 2 minutes; water rinse",
    "Counterstain with MALACHITE GREEN (or methylene blue) — 2 minutes; water rinse",
    "Dry at 60°C for ~5 minutes; mount with coverslip",
    "Examine 40x or 100x oil immersion; scan 200-300 fields",
  ];
  kSteps.forEach((st,i) => stepDot(s, i+1, st, 2.92, 2.31+i*0.43, 6.81, C.gi));

  // Result bar
  s.addShape(pres.shapes.RECTANGLE, {x:0.28, y:5.2, w:9.45, h:0.3, fill:{color:"E8F5EE"}, line:{color:C.gi,width:0.75}});
  s.addText("RESULTS: Oocysts = BRIGHT PINK-RED (acid-fast) | Background = BLUE-GREEN (malachite) | No heating needed = COLD method",
    {x:0.36, y:5.2, w:9.28, h:0.3, fontSize:9, color:C.gi, bold:true, valign:"middle"});
}

// ════════════════════════════════════════════
// SLIDE 6 — RESPIRATORY SPECIMENS + SMEAR PREP
// ════════════════════════════════════════════
{
  const s = pres.addSlide();
  bg(s, C.off);
  hdr(s, "Respiratory Tract — Specimens & Smear Preparation", C.resp);

  // Specimen table
  colHdr(s, 0.28, 1.05, 4.55, 0.32, "RESPIRATORY SPECIMENS", C.resp);
  tbl(s,
    ["Specimen","Collection Method","Used For"],
    [
      ["Expectorated sputum","Deep cough after rinsing mouth","Bacteria, TB, fungi"],
      ["Induced sputum","Inhale hypertonic saline (3-5%)","TB, PCP (can't produce sputum)"],
      ["NP swab","Flocked swab to posterior pharynx","Viruses, Bordetella, Mycoplasma"],
      ["BAL","Bronchoscopy, 60-120 mL saline","TB, PCP, fungi (immunocomp.)"],
      ["ET aspirate","Via endotracheal tube","Ventilator-assoc. pneumonia"],
    ],
    0.28, 1.4, 4.55, [1.35, 1.9, 1.3]
  );

  // Smear prep steps
  colHdr(s, 5.0, 1.05, 4.73, 0.32, "SPUTUM SMEAR PREPARATION", C.resp);
  const respSteps = [
    "Select PURULENT (green/yellow) portion — reject watery/frothy portions (saliva)",
    "Transfer small amount to center of clean, grease-free, labeled glass slide",
    "Spread THINLY and EVENLY over ~2×3 cm in one smooth stroke",
    "AIR-DRY completely at room temperature (5–10 minutes)",
    "HEAT-FIX: pass through flame 2-3× (1 sec each) OR hot plate 60°C for 5-10 min",
    "For BAL: use CYTOCENTRIFUGE (cytospin) for monolayer preparation",
    "Quality check: <10 squamous cells + >25 PMNs per 10x field (Bartlett criteria)",
  ];
  respSteps.forEach((st,i) => stepDot(s, i+1, st, 5.0, 1.4+i*0.53, 4.73, C.resp));

  s.addShape(pres.shapes.RECTANGLE, {x:5.0, y:5.16, w:4.73, h:0.3, fill:{color:"FFF3E0"}, line:{color:C.resp,width:0.75}});
  s.addText("⚠ BARTLETT CRITERION: <10 squamous cells + >25 PMNs/LPF = acceptable sputum (lower respiratory, not saliva)",
    {x:5.08, y:5.16, w:4.58, h:0.3, fontSize:8.5, color:C.resp, bold:true, valign:"middle"});
}

// ════════════════════════════════════════════
// SLIDE 7 — GRAM STAIN (DETAILED)
// ════════════════════════════════════════════
{
  const s = pres.addSlide();
  bg(s, C.off);
  hdr(s, "Gram Stain — Principle, Procedure & Results", C.resp);

  // Principle banner
  s.addShape(pres.shapes.RECTANGLE, {x:0.28, y:1.07, w:9.45, h:0.42, fill:{color:"FFF3E0"}, line:{color:C.resp,width:0.5}});
  s.addText("PRINCIPLE: Gram+ bacteria have THICK peptidoglycan walls — retain crystal violet-iodine complex after decolorization → PURPLE. Gram− bacteria have THIN peptidoglycan + lipopolysaccharide outer membrane — lose crystal violet → stained by safranin → PINK.",
    {x:0.36, y:1.07, w:9.28, h:0.42, fontSize:9.5, color:C.text, valign:"middle"});

  // Procedure table
  colHdr(s, 0.28, 1.55, 6.3, 0.32, "PROCEDURE (Reagent → Time)", C.resp);
  tbl(s,
    ["Step","Reagent","Time","Action"],
    [
      ["1","Crystal violet (primary stain)","1 min","Floods smear — ALL cells turn purple"],
      ["2","Water wash","2 sec","Removes excess stain"],
      ["3","Gram's iodine (mordant)","1 min","Forms crystal violet-iodine complex"],
      ["4","Water wash","2 sec","—"],
      ["5 ⚠","95% ethanol / acetone (decolorizer)","15-30 sec","CRITICAL — run until colorless runoff"],
      ["6","Water wash IMMEDIATELY","—","Stops decolorization"],
      ["7","Safranin (counterstain)","30-60 sec","Stains Gram− cells pink/red"],
      ["8","Water wash; blot dry","—","Examine 100x oil immersion"],
    ],
    0.28, 1.9, 6.3, [0.4, 2.3, 1.2, 2.4]
  );

  // Results panel
  colHdr(s, 6.75, 1.55, 2.98, 0.32, "RESULTS", C.resp);
  const res = [
    ["Gram + (PURPLE)","Strep pneumoniae, Staph aureus"],
    ["Gram − (PINK)","E. coli, Klebsiella, Pseudomonas"],
    ["Lancet diplococci","Streptococcus pneumoniae"],
    ["Clusters (cocci)","Staphylococcus aureus"],
    ["Small coccobacilli","Haemophilus influenzae"],
    ["Intracellular diplococci","N. gonorrhoeae (urethral)"],
    ["Squamous cells >10","Reject — saliva specimen"],
  ];
  res.forEach((r,i) => {
    const fy = 1.9 + i*0.38;
    s.addShape(pres.shapes.RECTANGLE, {x:6.75, y:fy, w:2.98, h:0.36, fill:{color:i%2===0?C.off:C.white}, line:{color:C.silver,width:0.5}});
    s.addText(r[0], {x:6.82, y:fy+0.01, w:2.82, h:0.18, fontSize:8.5, bold:true, color:C.resp});
    s.addText(r[1], {x:6.82, y:fy+0.19, w:2.82, h:0.15, fontSize:8, color:C.text});
  });

  // Memory Aid
  s.addShape(pres.shapes.ROUNDED_RECTANGLE, {x:0.28, y:4.82, w:6.3, h:0.66, fill:{color:C.navy}, line:{color:C.navy,width:0}, rectRadius:0.1});
  s.addText("MEMORY AID — GRAM STAIN SEQUENCE:\n\"CIDS\" → Crystal violet → Iodine → Decolorize → Safranin",
    {x:0.38, y:4.82, w:6.1, h:0.66, fontSize:11, bold:true, color:C.yellow, align:"center", valign:"middle"});
}

// ════════════════════════════════════════════
// SLIDE 8 — ZIEHL-NEELSEN (ZN) STAIN
// ════════════════════════════════════════════
{
  const s = pres.addSlide();
  bg(s, C.off);
  hdr(s, "Ziehl-Neelsen (ZN) Stain — Acid-Fast Bacteria", C.resp);

  // Principle
  s.addShape(pres.shapes.RECTANGLE, {x:0.28, y:1.05, w:9.45, h:0.4, fill:{color:"FFF3E0"}, line:{color:C.resp,width:0.5}});
  s.addText("PRINCIPLE: Mycobacteria have HIGH MYCOLIC ACID content in cell wall → resistant to decolorization. HEAT drives carbol fuchsin into the waxy wall. Once stained, AFB resist acid-alcohol decolorization → appear PINK-RED (acid-fast).",
    {x:0.36, y:1.05, w:9.28, h:0.4, fontSize:9.5, color:C.text, valign:"middle"});

  // Steps and WHO grading side by side
  colHdr(s, 0.28, 1.52, 5.6, 0.32, "HOT ACID-FAST PROCEDURE (Step-by-Step)", C.resp);
  const znSteps = [
    "Prepare smear; dry THOROUGHLY; heat-fix (incomplete drying = poor AFB staining)",
    "Flood with CARBOL FUCHSIN",
    "HEAT under the slide until STEAM RISES — maintain 5 minutes (do NOT boil); reapply stain as it dries",
    "COOL for 5 minutes; wash with distilled water",
    "Decolorize with ACID-ALCOHOL (3% HCl in 95% ethanol) until smear is pale pink — 2-3 min; wash",
    "Counterstain with METHYLENE BLUE — 1-2 minutes; wash; blot dry",
    "Examine 100x oil immersion; scan 100-300 fields",
  ];
  znSteps.forEach((st,i) => stepDot(s, i+1, st, 0.28, 1.87+i*0.5, 5.6, C.resp));

  // WHO Grading table
  colHdr(s, 6.05, 1.52, 3.68, 0.32, "WHO GRADING (ZN Smear)", C.resp);
  tbl(s,
    ["Grade","Criteria"],
    [
      ["No AFB","0 AFB in 100 fields"],
      ["Scanty","1-9 AFB in 100 fields"],
      ["1+","10-99 AFB in 100 fields"],
      ["2+","1-10 AFB/field (50 fields)"],
      ["3+",">10 AFB/field (20 fields)"],
    ],
    6.05, 1.87, 3.68, [1.0, 2.68]
  );

  // Memory + Results
  s.addShape(pres.shapes.RECTANGLE, {x:6.05, y:3.72, w:3.68, h:0.56, fill:{color:"FFF3E0"}, line:{color:C.resp,width:0.75}});
  s.addText("RESULTS:\n• AFB → BRIGHT PINK-RED RODS\n• Background → BLUE",
    {x:6.12, y:3.72, w:3.54, h:0.56, fontSize:9.5, color:C.resp, bold:true, valign:"middle"});

  s.addShape(pres.shapes.ROUNDED_RECTANGLE, {x:0.28, y:4.75, w:5.6, h:0.72, fill:{color:C.navy}, line:{color:C.navy,width:0}, rectRadius:0.1});
  s.addText("MEMORY AID — ZN SEQUENCE:\n\"CAM\" → Carbol fuchsin (+ heat) → Acid-alcohol → Methylene blue",
    {x:0.38, y:4.75, w:5.4, h:0.72, fontSize:11, bold:true, color:C.yellow, align:"center", valign:"middle"});

  s.addShape(pres.shapes.ROUNDED_RECTANGLE, {x:6.05, y:4.38, w:3.68, h:1.09, fill:{color:C.navy}, line:{color:C.navy,width:0}, rectRadius:0.1});
  s.addText("ZN vs. AURAMINE:\nZN = Hot, morphology details\nAuramine = Cold fluorescence, MORE SENSITIVE for screening\n→ Auramine positives must be confirmed by ZN",
    {x:6.12, y:4.38, w:3.54, h:1.09, fontSize:9, color:C.silver, valign:"middle"});
}

// ════════════════════════════════════════════
// SLIDE 9 — URINARY TRACT SPECIMENS + PREP
// ════════════════════════════════════════════
{
  const s = pres.addSlide();
  bg(s, C.off);
  hdr(s, "Urinary Tract — Specimens & Smear Preparation", C.urine);

  // Specimen table left
  colHdr(s, 0.28, 1.05, 4.55, 0.32, "URINE SPECIMENS", C.urine);
  tbl(s,
    ["Specimen","Method","Used For"],
    [
      ["MSCC (midstream clean catch)","Patient self-collects","Routine culture, microscopy"],
      ["Catheter specimen (CSU)","From catheter port — NOT bag","Hospital inpatients"],
      ["Suprapubic aspirate (SPA)","Needle directly into bladder","Gold standard; infants"],
      ["Early morning urine","First void — concentrated","TB (AFB), best for microscopy"],
      ["Urethral swab/discharge","Swab 2-4 cm into urethra","GC / Chlamydia"],
    ],
    0.28, 1.4, 4.55, [1.5, 1.75, 1.3]
  );

  // Smear prep right
  colHdr(s, 5.0, 1.05, 4.73, 0.32, "URINE SMEAR PREPARATION", C.urine);
  colHdr(s, 5.0, 1.4, 4.73, 0.28, "Method 1: Centrifuged Sediment (Standard)", C.urine);
  const m1 = [
    "Pour 10 mL urine into conical tube",
    "Centrifuge 2000 rpm (400×g) for 5 minutes",
    "Decant supernatant; leave ~0.5-1 mL sediment",
    "Resuspend sediment by tapping tube",
    "Place ONE DROP on glass slide",
    "Wet mount → coverslip; examine immediately | Stained → air-dry; heat-fix",
  ];
  m1.forEach((st,i) => stepDot(s, i+1, st, 5.0, 1.7+i*0.45, 4.73, C.urine));

  colHdr(s, 5.0, 3.72, 4.73, 0.28, "Method 2: Uncentrifuged (Gram stain screening)", C.urine);
  const m2 = [
    "Place drop of well-mixed uncentrifuged urine on slide",
    "Spread thinly; air-dry; heat-fix → proceed to Gram stain",
    "1 organism/oil-immersion field ≈ 100,000 CFU/mL (significant bacteriuria)",
  ];
  m2.forEach((st,i) => stepDot(s, i+1, st, 5.0, 4.02+i*0.46, 4.73, C.urine));

  s.addShape(pres.shapes.RECTANGLE, {x:5.0, y:5.42, w:4.73, h:0.26, fill:{color:"F3EEF8"}, line:{color:C.urine,width:0.75}});
  s.addText("⚠  Early morning urine = examine within MINUTES (cells lyse in dilute/alkaline urine; Trichomonas motility lost rapidly)",
    {x:5.08, y:5.42, w:4.58, h:0.26, fontSize:8.5, color:C.urine, bold:true, valign:"middle"});
}

// ════════════════════════════════════════════
// SLIDE 10 — URINE STAINING: WET MOUNT + GRAM
// ════════════════════════════════════════════
{
  const s = pres.addSlide();
  bg(s, C.off);
  hdr(s, "Urinary Tract — Wet Mount & Gram Stain", C.urine);

  // Wet mount left
  colHdr(s, 0.28, 1.05, 4.55, 0.32, "WET MOUNT OF URINE SEDIMENT (Unstained)", C.urine);
  const wmItems = [
    ["Trichomonas vaginalis","Pear-shaped, jerky motility with flagella — examine IMMEDIATELY"],
    ["RBCs","Normal: 0-2/HPF; >3/HPF = hematuria"],
    ["WBCs (pus cells)",">5/HPF = pyuria (UTI, pyelonephritis)"],
    ["RBC casts","Glomerulonephritis"],
    ["WBC casts","Pyelonephritis (upper UTI)"],
    ["Squamous cells","Contamination (vaginal/skin)"],
    ["Yeast (Candida)","Oval budding cells + pseudohyphae"],
  ];
  wmItems.forEach((r,i) => {
    const y = 1.4 + i*0.48;
    s.addShape(pres.shapes.RECTANGLE, {x:0.28, y, w:4.55, h:0.46, fill:{color:i%2===0?C.off:C.white}, line:{color:C.silver,width:0.5}});
    s.addText(r[0], {x:0.36, y:y+0.02, w:4.38, h:0.2, fontSize:9, bold:true, color:C.urine});
    s.addText(r[1], {x:0.36, y:y+0.22, w:4.38, h:0.2, fontSize:8.5, color:C.text});
  });

  // Gram stain for urine right
  colHdr(s, 5.0, 1.05, 4.73, 0.32, "GRAM STAIN OF URINE — Key Findings", C.urine);
  s.addText("Steps: SAME as respiratory Gram stain\n(Crystal violet → Iodine → Decolorize → Safranin)",
    {x:5.0, y:1.4, w:4.73, h:0.44, fontSize:9.5, color:C.text, valign:"middle", italic:true});
  tbl(s,
    ["Organism","Gram Reaction","Clinical Significance"],
    [
      ["Escherichia coli","Gram − rods","Most common UTI (80%)"],
      ["Klebsiella pneumoniae","Gram − rods","Complicated UTI"],
      ["Staphylococcus saprophyticus","Gram + cocci","UTI in young women"],
      ["Enterococcus faecalis","Gram + cocci (chains)","UTI, nosocomial"],
      ["N. gonorrhoeae","Gram − diplococci (INTRACELLULAR in PMNs)","Urethral discharge"],
      ["Candida spp.","Gram + yeast + pseudohyphae","Immunocompromised, catheter"],
    ],
    5.0, 1.87, 4.73, [1.5, 1.6, 1.63]
  );

  // Kass criteria
  s.addShape(pres.shapes.ROUNDED_RECTANGLE, {x:5.0, y:4.82, w:4.73, h:0.72, fill:{color:C.navy}, line:{color:C.navy,width:0}, rectRadius:0.1});
  s.addText("KASS CRITERIA (Significant Bacteriuria):\n≥10⁵ CFU/mL (MSCC) | ≥10³ CFU/mL (catheter) | ANY growth (SPA)",
    {x:5.08, y:4.82, w:4.58, h:0.72, fontSize:10.5, bold:true, color:C.yellow, align:"center", valign:"middle"});
}

// ════════════════════════════════════════════
// SLIDE 11 — ZN STAIN FOR URINE (RENAL TB)
// ════════════════════════════════════════════
{
  const s = pres.addSlide();
  bg(s, C.off);
  hdr(s, "Urinary Tract — ZN Stain for Renal TB + Comparison", C.urine);

  // Left — ZN urine
  colHdr(s, 0.28, 1.05, 4.55, 0.32, "ZIEHL-NEELSEN FOR URINE (Renal TB)", C.urine);
  const zuSteps = [
    "Collect EARLY MORNING midstream urine × 3 CONSECUTIVE DAYS",
    "(Mycobacteria shed intermittently — 3 samples increases sensitivity)",
    "Centrifuge LARGE volume (50-100 mL) at high speed to concentrate",
    "Prepare thin smear from sediment; air-dry; heat-fix",
    "Flood with carbol fuchsin; heat until steam rises — 5 minutes",
    "Cool; wash; acid-alcohol decolorize (3% HCl) — 2-3 min; wash",
    "Counterstain methylene blue — 1-2 min; wash; blot dry",
    "Examine 100x oil; grade by WHO criteria",
  ];
  zuSteps.forEach((st,i) => stepDot(s, i+1, st, 0.28, 1.4+i*0.5, 4.55, C.urine));

  // Right — ZN vs hot vs cold comparison
  colHdr(s, 5.0, 1.05, 4.73, 0.32, "ZN HOT vs KINYOUN COLD — Comparison", C.urine);
  tbl(s,
    ["Feature","ZN (Hot)","Kinyoun (Cold)"],
    [
      ["Heat required","YES — steam required","NO"],
      ["Primary target","Mycobacteria (AFB)","Coccidian oocysts"],
      ["Decolorizer","Acid-alcohol (3% HCl)","Acid-alcohol (1% HCl)"],
      ["Counterstain","Methylene blue","Malachite green"],
      ["Sensitivity for AFB","Gold standard","Less sensitive for AFB"],
      ["Used in","Sputum, urine, gastric","Stool, BAL"],
    ],
    5.0, 1.4, 4.73, [1.5, 1.62, 1.61]
  );

  // Alert + result
  s.addShape(pres.shapes.RECTANGLE, {x:0.28, y:5.42, w:9.45, h:0.3, fill:{color:"F3EEF8"}, line:{color:C.urine,width:0.75}});
  s.addText("RESULTS — AFB = PINK-RED rods on BLUE background  |  3 early morning urines must be submitted for renal TB diagnosis",
    {x:0.36, y:5.42, w:9.28, h:0.3, fontSize:9, color:C.urine, bold:true, valign:"middle"});
}

// ════════════════════════════════════════════
// SLIDE 12 — MASTER COMPARISON TABLE
// ════════════════════════════════════════════
{
  const s = pres.addSlide();
  bg(s, C.off);
  hdr(s, "Master Comparison — All Stains, All Three Tracts (RGUHS Exam)", C.navy);

  tbl(s,
    ["Stain","Principle","Reagents (in order)","Result","Tract(s)","★ RGUHS"],
    [
      ["Wet Mount","Direct motility/morphology","Saline / Diluted Lugol's iodine","Motile protozoa; cysts (iodine)","GI, Urine","★★★"],
      ["Gram Stain","Cell wall thickness (peptidoglycan)","Crystal violet → Iodine → EtOH → Safranin","G+ = purple | G− = pink","ALL 3","★★★★★"],
      ["ZN (Hot)","Mycolic acid; acid-fastness","Carbol fuchsin (+heat) → Acid-alcohol → Methylene blue","AFB = pink-red | Bg = blue","GI, Resp, Urine","★★★★★"],
      ["Kinyoun (Cold)","Acid-fastness; oocysts","Carbol fuchsin → Acid-alcohol → Malachite green","Oocysts = pink-red | Bg = blue-green","GI, BAL","★★★"],
      ["Trichrome (Wheatley)","Differential dye affinity","EtOH-iodine → Trichrome → Acid-EtOH → Xylene","Protozoa = blue-green/purple | Bg = green","GI","★★★★"],
      ["Auramine-Phenol","Fluorochrome binds mycolic acid","Auramine → Acid-alcohol → KMnO₄","AFB = yellow-green (fluorescence)","Resp","★★★"],
      ["GMS","Methenamine silver on cell walls","Oxidize → Methenamine silver → Light green","Fungi/PCP = black | Bg = green","Resp","★★"],
      ["Pap Stain","Differential cytoplasm staining","Hematoxylin → Orange G → Eosin Azure","Nuclei = blue | Cytoplasm = pink","Resp, Urine","★★"],
    ],
    0.18, 1.12, 9.64, [1.5, 1.6, 2.35, 1.65, 1.0, 1.54]
  );
}

// ════════════════════════════════════════════
// SLIDE 13 — EXAM QUESTIONS + MEMORY AIDS
// ════════════════════════════════════════════
{
  const s = pres.addSlide();
  bg(s, C.off);
  hdr(s, "RGUHS Exam Questions & Memory Aids", C.navy);

  // Left — exam questions
  colHdr(s, 0.28, 1.05, 5.3, 0.32, "IMPORTANT RGUHS EXAM QUESTIONS", C.navy);
  card(s, 0.28, 1.4, 5.3, 3.9);

  s.addText("LONG ANSWER (10 marks):", {x:0.38, y:1.46, w:5.1, h:0.3, fontSize:10, bold:true, color:C.resp});
  const lq = [
    "• Smear preparation and Gram staining of sputum with results",
    "• Describe ZN staining — principle, procedure, results",
    "• Specimen collection, smear prep and staining for respiratory tract infections",
  ];
  lq.forEach((q,i) => s.addText(q, {x:0.38, y:1.78+i*0.36, w:5.1, h:0.34, fontSize:9.5, color:C.text}));

  s.addText("SHORT ANSWER (5 marks):", {x:0.38, y:2.9, w:5.1, h:0.3, fontSize:10, bold:true, color:C.resp});
  const sq = [
    "• Wheatley trichrome stain | • Modified Kinyoun stain",
    "• Bartlett criteria for sputum quality",
    "• Kass criteria for significant bacteriuria",
    "• WHO grading of ZN smear | • Auramine vs ZN",
  ];
  sq.forEach((q,i) => s.addText(q, {x:0.38, y:3.22+i*0.35, w:5.1, h:0.33, fontSize:9.5, color:C.text}));

  s.addText("VERY SHORT ANSWER (2 marks):", {x:0.38, y:4.66, w:5.1, h:0.28, fontSize:10, bold:true, color:C.resp});
  s.addText("• Reagents of Gram stain in order  • Why heat in ZN?  • What does 1 org/OIF mean in urine?  • Two permanent stains for fecal specimens",
    {x:0.38, y:4.96, w:5.1, h:0.3, fontSize:9, color:C.text});

  // Right — memory aids
  colHdr(s, 5.75, 1.05, 3.98, 0.32, "MEMORY AIDS", C.navy);
  const aids = [
    {title:"Gram Stain — \"CIDS\"", body:"Crystal violet → Iodine → Decolorize → Safranin"},
    {title:"ZN Stain — \"CAM\"", body:"Carbol fuchsin (+heat) → Acid-alcohol → Methylene blue"},
    {title:"UTI Organisms — \"KEEPS\"", body:"Klebsiella, E. coli, Enterococcus, Pseudomonas, Staph"},
    {title:"Kass Criteria", body:"10⁵/mL (MSCC) | 10³/mL (catheter) | Any (SPA)"},
    {title:"Bartlett Criteria", body:"<10 squamous cells + >25 PMNs per LPF"},
    {title:"ZN vs Kinyoun", body:"ZN = HOT (steam); Kinyoun = COLD (no heat)"},
  ];
  aids.forEach((a,i) => {
    const y = 1.4 + i*0.69;
    s.addShape(pres.shapes.ROUNDED_RECTANGLE, {x:5.75, y, w:3.98, h:0.65, fill:{color:i%2===0?C.navy:"142850"}, line:{color:C.teal,width:0.75}, rectRadius:0.1});
    s.addText(a.title, {x:5.83, y:y+0.02, w:3.82, h:0.26, fontSize:9.5, bold:true, color:C.yellow});
    s.addText(a.body, {x:5.83, y:y+0.28, w:3.82, h:0.3, fontSize:9, color:C.silver});
  });
}

// ════════════════════════════════════════════
// SLIDE 14 — CONCLUSION / KEY TAKEAWAYS
// ════════════════════════════════════════════
{
  const s = pres.addSlide();
  bg(s, C.navy);
  s.addShape(pres.shapes.RECTANGLE, {x:0, y:4.5, w:"100%", h:1.125, fill:{color:C.teal}, line:{color:C.teal,width:0}});
  s.addShape(pres.shapes.RECTANGLE, {x:0, y:3.65, w:"100%", h:0.07, fill:{color:C.yellow}, line:{color:C.yellow,width:0}});

  s.addText("KEY POINTS TO REMEMBER", {x:0.5, y:0.2, w:9, h:0.62, fontSize:26, bold:true, color:C.teal, align:"center", charSpacing:4});

  const pts = [
    {n:"1",t:"Smear QUALITY = thin, even, properly fixed. Thick smears are the most common error in the lab."},
    {n:"2",t:"GRAM STAIN is first-line for ALL three tracts. Sequence: CIDS (Crystal violet → Iodine → Decolorize → Safranin)."},
    {n:"3",t:"GI: Wet mount (motility) + Trichrome (protozoa) + Modified Kinyoun (Cryptosporidium/Cyclospora/Isospora)."},
    {n:"4",t:"RESPIRATORY: Check sputum quality (Bartlett criteria). ZN = CAM (steam required). Auramine is more sensitive but must confirm with ZN."},
    {n:"5",t:"URINARY: Centrifuge 10 mL at 2000 rpm. Wet mount IMMEDIATELY for Trichomonas. Kass: ≥10⁵ CFU/mL = significant bacteriuria."},
    {n:"6",t:"RENAL TB: 3 early morning urines × 3 consecutive days. Large volume centrifugation to concentrate AFB."},
  ];
  pts.forEach((p,i) => {
    const col = i%2; const row = Math.floor(i/2);
    const x = 0.3 + col*4.87; const y = 1.0 + row*1.06;
    s.addShape(pres.shapes.OVAL, {x, y, w:0.36, h:0.36, fill:{color:C.yellow}, line:{color:C.yellow,width:0}});
    s.addText(p.n, {x, y, w:0.36, h:0.36, fontSize:11, bold:true, color:C.navy, align:"center", valign:"middle", margin:0});
    s.addText(p.t, {x:x+0.44, y:y+0.02, w:4.22, h:0.88, fontSize:9.5, color:C.silver, fontFace:"Calibri", valign:"top"});
  });

  s.addText("3rd B.Sc. MLT  |  RGUHS Microbiology II  |  Specimen Processing & Staining", {x:0.5, y:4.6, w:9, h:0.44, fontSize:11, color:C.navy, align:"center", italic:true, bold:true, valign:"middle"});
}

// ── WRITE ─────────────────────────────────
pres.writeFile({fileName:"/home/daytona/workspace/rguhs-ppt/RGUHS_3rd_BSc_MLT_Smear_and_Staining.pptx"})
  .then(()=>console.log("✅ Done!"))
  .catch(e=>console.error("❌",e));

cd /home/daytona/workspace/rguhs-ppt && node create_rguhs_ppt.js

ls -lh /home/daytona/workspace/rguhs-ppt/

RGUHS 3rd BSc MLT Pathology II syllabus smear preparation staining GI respiratory urinary tract histopathology cytology

https://www.gitam.edu/sites/default/files/syllabus/BSc_Medic…

RGUHS BSc MLT pathology smear preparation staining Papanicolaou HE MGG respiratory urinary GI tract exfoliative cytology

~/rguhs-path-ppt/create_path_ppt.js

const pptxgen = require("pptxgenjs");
const pres = new pptxgen();
pres.layout = "LAYOUT_16x9";
pres.title = "Smear Preparation and Staining - Pathology - 3rd BSc MLT RGUHS";

const C = {
  navy:   "0B2545",
  teal:   "0E7C7C",
  white:  "FFFFFF",
  off:    "F0F4F8",
  silver: "C8D6E5",
  text:   "1A1A2E",
  gi:     "1A6B3A",
  resp:   "A63D00",
  urine:  "55178A",
  yellow: "F4C430",
  purple: "6A0DAD",
  ltgray: "E8EEF4",
  fix:    "1A4A7A",
};

function bg(s,c){ s.addShape(pres.shapes.RECTANGLE,{x:0,y:0,w:"100%",h:"100%",fill:{color:c},line:{color:c,width:0}}); }
function hdr(s,t,c){
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}
function colHdr(s,x,y,w,h,t,c){
  s.addShape(pres.shapes.RECTANGLE,{x,y,w,h,fill:{color:c},line:{color:c,width:0}});
  s.addText(t,{x,y,w,h,fontSize:9,bold:true,color:C.white,align:"center",valign:"middle",margin:0});
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function stepDot(s,n,t,x,y,w,ac){
  s.addShape(pres.shapes.OVAL,{x:x+0.05,y:y+0.07,w:0.29,h:0.29,fill:{color:ac},line:{color:ac,width:0}});
  s.addText(String(n),{x:x+0.05,y:y+0.07,w:0.29,h:0.29,fontSize:8.5,bold:true,color:C.white,align:"center",valign:"middle",margin:0});
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function tbl(s,headers,rows,x,y,w,colW){
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}
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}

// ── SLIDE 1: TITLE ────────────────────────────────────
{
  const s=pres.addSlide();
  bg(s,C.navy);
  s.addShape(pres.shapes.RECTANGLE,{x:0,y:3.5,w:"100%",h:0.07,fill:{color:C.teal},line:{color:C.teal,width:0}});
  s.addShape(pres.shapes.RECTANGLE,{x:0,y:3.58,w:"100%",h:0.04,fill:{color:C.yellow},line:{color:C.yellow,width:0}});

  s.addText("SMEAR PREPARATION",{x:0.5,y:0.4,w:9,h:0.82,fontSize:38,bold:true,color:C.white,align:"center",charSpacing:3,fontFace:"Calibri"});
  s.addText("& STAINING PROCEDURES",{x:0.5,y:1.2,w:9,h:0.72,fontSize:32,bold:true,color:C.teal,align:"center",charSpacing:2,fontFace:"Calibri"});

  s.addShape(pres.shapes.ROUNDED_RECTANGLE,{x:2.7,y:2.08,w:4.6,h:1.18,fill:{color:"102040"},line:{color:C.teal,width:1.5},rectRadius:0.12});
  s.addText("3rd B.Sc. MLT  |  RGUHS\nPathology — Cytopathology\n(Exfoliative Cytology)",{x:2.7,y:2.08,w:4.6,h:1.18,align:"center",valign:"middle",
    fontSize:12,bold:true,color:C.yellow,fontFace:"Calibri"});

  const pills=[{l:"GASTROINTESTINAL",c:C.gi,x:0.4},{l:"RESPIRATORY TRACT",c:C.resp,x:3.65},{l:"URINARY TRACT",c:C.urine,x:6.9}];
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    s.addShape(pres.shapes.ROUNDED_RECTANGLE,{x:p.x,y:3.68,w:2.75,h:0.55,fill:{color:p.c},line:{color:p.c,width:0},rectRadius:0.27});
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  s.addText("Stains Covered: Papanicolaou (Pap)  |  May-Grünwald Giemsa (MGG)  |  Haematoxylin & Eosin (H&E)",{x:0.5,y:4.9,w:9,h:0.35,fontSize:10,color:C.silver,align:"center",italic:true});
}

// ── SLIDE 2: EXFOLIATIVE CYTOLOGY + FIXATION ─────────
{
  const s=pres.addSlide();
  bg(s,C.off);
  hdr(s,"Exfoliative Cytology — Introduction & Fixation",C.navy);

  colHdr(s,0.28,1.05,4.55,0.32,"EXFOLIATIVE CYTOLOGY",C.navy);
  const defPoints=[
    ["Definition","Microscopic study of cells shed spontaneously or scraped from body surfaces/cavities"],
    ["Types","(1) Exfoliative — spontaneously shed (sputum, urine, vaginal secretion)\n(2) Interventional — collected by procedure (brushing, washing, FNAC)"],
    ["Advantages","Non-invasive, rapid, inexpensive, repeatable"],
    ["Disadvantages","Cannot assess tissue architecture; sampling errors possible"],
    ["Specimens","Sputum, urine, gastric washings, brushings, body fluids, cervical smear"],
  ];
  defPoints.forEach(([k,v],i)=>{
    const y=1.4+i*0.72;
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  colHdr(s,5.0,1.05,4.73,0.32,"FIXATION — TYPES & USES",C.fix);
  tbl(s,
    ["Fixative","Type","Used For"],
    [
      ["95% Ethyl alcohol","Wet fixation","Pap stain (gold standard)"],
      ["Equal parts 95% EtOH + Ether","Wet fixation","Papanicolaou original fixative"],
      ["Spray fixative (Cytosprep)","Wet fixation","Field use — Pap stain"],
      ["Air drying","Dry fixation","MGG stain — MUST air dry"],
      ["Absolute methanol","Dry/wet","MGG, H&E smears"],
    ],
    5.0,1.4,4.73,[1.8,1.35,1.58]
  );
  s.addShape(pres.shapes.RECTANGLE,{x:5.0,y:4.48,w:4.73,h:0.64,fill:{color:"EBF4FF"},line:{color:C.fix,width:0.75}});
  s.addText("KEY RULE:\n• WET fixation (alcohol) = Pap & H&E → fix BEFORE drying\n• AIR DRYING = MGG → must dry BEFORE staining\n⚠ Wrong fixation = RUINED smear!",
    {x:5.08,y:4.48,w:4.58,h:0.64,fontSize:9.5,color:C.fix,fontFace:"Calibri",valign:"top",bold:true});
}

// ── SLIDE 3: RESPIRATORY SPECIMENS + SMEAR PREP ───────
{
  const s=pres.addSlide();
  bg(s,C.off);
  hdr(s,"Respiratory Tract — Specimens & Smear Preparation",C.resp);

  colHdr(s,0.28,1.05,4.55,0.32,"RESPIRATORY SPECIMENS (Cytology)",C.resp);
  tbl(s,
    ["Specimen","Collection","Purpose"],
    [
      ["Sputum","Deep cough after rinsing mouth; early morning (×3 for TB)","Malignancy, infection, TB"],
      ["Bronchial washing","Saline via bronchoscope; aspirated","Peripheral lung lesions"],
      ["Bronchial brushing","Brush via bronchoscope; smear direct","Bronchial carcinoma"],
      ["BAL","60-120 mL saline via bronchoscope","PCP, TB, infections, ILD"],
      ["Pleural fluid","Thoracentesis aspirate","Effusions, malignancy"],
    ],
    0.28,1.4,4.55,[1.35,1.8,1.4]
  );

  colHdr(s,5.0,1.05,4.73,0.32,"SPUTUM SMEAR PREPARATION",C.resp);
  const rsteps=[
    "Handle in Class II biosafety cabinet (infectious aerosol risk)",
    "Select the MOST PURULENT / blood-stained portion using applicator sticks",
    "Spread THINLY and EVENLY on 2-3 glass slides (circular or longitudinal)",
    "For PAP: Fix IMMEDIATELY in 95% ethanol (before it dries)",
    "For MGG: Allow to AIR DRY completely at room temperature",
    "Label slides with patient ID, date, specimen type",
    "For BAL/washings: Centrifuge 1500-2000 rpm × 10 min; use sediment OR cytospin",
  ];
  rsteps.forEach((st,i)=>stepDot(s,i+1,st,5.0,1.4+i*0.54,4.73,C.resp));
  s.addShape(pres.shapes.RECTANGLE,{x:5.0,y:5.2,w:4.73,h:0.3,fill:{color:"FFF3E0"},line:{color:C.resp,width:0.75}});
  s.addText("⚠  Cytocentrifuge (cytospin) gives a monolayer — preferred for washings/BAL; better cellular morphology",
    {x:5.08,y:5.2,w:4.58,h:0.3,fontSize:8.5,color:C.resp,bold:true,valign:"middle"});
}

// ── SLIDE 4: PAP STAIN PROCEDURE ─────────────────────
{
  const s=pres.addSlide();
  bg(s,C.off);
  hdr(s,"Papanicolaou (Pap) Stain — Principle, Procedure & Results",C.navy);

  // Principle banner
  s.addShape(pres.shapes.RECTANGLE,{x:0.28,y:1.05,w:9.45,h:0.42,fill:{color:"EBF4FF"},line:{color:C.navy,width:0.5}});
  s.addText("PRINCIPLE: Multi-chromatic stain with 5 dyes in 3 solutions. Hematoxylin stains nuclei. OG-6 stains keratin/cornified cells. EA (Eosin Azure) stains cytoplasm polychromatically. Wet fixation (95% ethanol) ESSENTIAL — preserves nuclear detail.",
    {x:0.36,y:1.05,w:9.28,h:0.42,fontSize:9.5,color:C.text,valign:"middle"});

  // Procedure table
  colHdr(s,0.28,1.52,6.5,0.32,"PROCEDURE (Steps)",C.navy);
  tbl(s,
    ["Step","Reagent","Time","Purpose"],
    [
      ["1","Rehydrate: 80%→70%→50% ethanol","2 min each","Remove fixative"],
      ["2","Harris HEMATOXYLIN","3-6 min","Stains nuclei blue-black"],
      ["3","Water rinse","2 min","Removes excess stain"],
      ["4","Acid-alcohol (1% HCl in 70% EtOH)","6-10 dips","Differentiation"],
      ["5","Scott's tap water / ammonia water (0.5%)","1-2 min","BLUING — nuclei turn blue"],
      ["6","50%→70%→80%→95% ethanol","2 min each","Dehydration"],
      ["7","ORANGE G-6 (OG-6)","2-3 min","Stains keratin orange"],
      ["8","95% ethanol ×2","2 min each","Rinse OG"],
      ["9","EOSIN AZURE (EA-36/EA-50)","3-5 min","Cytoplasm polychrome"],
      ["10","95%→100% ethanol ×2 → Xylene ×2","2 min each","Dehydrate, Clear"],
      ["11","Mount with DPX + coverslip","—","Permanent mount"],
    ],
    0.28,1.87,6.5,[0.35,2.55,1.1,2.5]
  );

  // Results panel
  colHdr(s,6.95,1.52,2.78,0.32,"RESULTS",C.navy);
  const res=[["Nuclei","Blue-black"],["Superficial cells","Orange/pink (OG-6)"],["Intermediate cells","Green-blue (EA)"],["Parabasal/metaplastic","Green (light green)"],
    ["Malignant cells","Hyperchromatic, irregular nuclei"],["RBCs","Orange-red"],["Mucus","Pale green"],["Keratin","Deep orange"]];
  res.forEach((r,i)=>{
    const y=1.87+i*0.38;
    s.addShape(pres.shapes.RECTANGLE,{x:6.95,y,w:2.78,h:0.36,fill:{color:i%2===0?C.off:C.white},line:{color:C.silver,width:0.5}});
    s.addText(r[0],{x:7.02,y:y+0.01,w:2.64,h:0.18,fontSize:8.5,bold:true,color:C.navy});
    s.addText("→ "+r[1],{x:7.02,y:y+0.19,w:2.64,h:0.14,fontSize:8,color:C.text,italic:true});
  });

  s.addShape(pres.shapes.ROUNDED_RECTANGLE,{x:0.28,y:4.88,w:6.5,h:0.6,fill:{color:C.navy},line:{color:C.navy,width:0},rectRadius:0.1});
  s.addText("MEMORY AID — Pap Reagents: \"H-A-B-OG-EA\" → Hematoxylin → Acid-alcohol → Bluing → OG-6 → Eosin Azure",
    {x:0.38,y:4.88,w:6.3,h:0.6,fontSize:11,bold:true,color:C.yellow,align:"center",valign:"middle"});
}

// ── SLIDE 5: MGG STAIN ───────────────────────────────
{
  const s=pres.addSlide();
  bg(s,C.off);
  hdr(s,"May-Grünwald Giemsa (MGG) Stain — Procedure & Results",C.navy);

  // Principle
  s.addShape(pres.shapes.RECTANGLE,{x:0.28,y:1.05,w:9.45,h:0.38,fill:{color:"EBF4FF"},line:{color:C.navy,width:0.5}});
  s.addText("PRINCIPLE: Romanowsky-type stain. Methylene blue (basic dye) stains acidic components (nuclei, RNA) blue/purple. Eosin (acidic dye) stains basic components (haemoglobin, eosinophil granules) pink/red. AIR-DRIED SMEARS only.",
    {x:0.36,y:1.05,w:9.28,h:0.38,fontSize:9.5,color:C.text,valign:"middle"});

  colHdr(s,0.28,1.48,5.4,0.32,"PROCEDURE (Step-by-Step)",C.purple);
  const mggSteps=[
    "Smear must be AIR-DRIED completely (CRITICAL — wet smear gives poor results)",
    "Fix in ABSOLUTE METHANOL — 3-5 minutes",
    "MAY-GRÜNWALD stain (undiluted) — 3 minutes",
    "Add EQUAL VOLUME of phosphate buffer (pH 6.8) to dilute stain on slide — 1 minute",
    "Wash off with phosphate buffer",
    "GIEMSA stain (diluted 1:20 in phosphate buffer pH 6.8) — 15-20 minutes",
    "Wash off with distilled water or buffer",
    "Air dry completely; mount with DPX + coverslip",
    "Examine 40x then 100x oil immersion",
  ];
  mggSteps.forEach((st,i)=>stepDot(s,i+1,st,0.28,1.83+i*0.415,5.4,C.purple));

  // Results + comparison with Pap
  colHdr(s,5.85,1.48,3.88,0.32,"RESULTS (MGG)",C.purple);
  const mggRes=[["Nuclei","Purple-blue / violet"],["Basophilic cytoplasm","Blue"],["Eosinophilic cytoplasm","Pink-red"],
    ["Eosinophil granules","Bright red-orange"],["Neutrophil granules","Purple"],["RBCs","Pink"],["Mucus/background","Pale pink"],["Lymphocytes","Deep purple nucleus"]];
  mggRes.forEach((r,i)=>{
    const y=1.83+i*0.41;
    s.addShape(pres.shapes.RECTANGLE,{x:5.85,y,w:3.88,h:0.39,fill:{color:i%2===0?C.off:C.white},line:{color:C.silver,width:0.5}});
    s.addText(r[0],{x:5.92,y:y+0.01,w:3.72,h:0.18,fontSize:8.5,bold:true,color:C.purple});
    s.addText("→ "+r[1],{x:5.92,y:y+0.2,w:3.72,h:0.16,fontSize:8,color:C.text,italic:true});
  });

  s.addShape(pres.shapes.ROUNDED_RECTANGLE,{x:0.28,y:5.0,w:9.45,h:0.48,fill:{color:"F5F0FF"},line:{color:C.purple,width:0.75},rectRadius:0.08});
  s.addText("KEY: MGG = AIR DRY first → then fix in methanol | PAP = Fix in 95% ethanol FIRST (before drying)  | MGG best for FNAC, lymphoma, background material",
    {x:0.38,y:5.0,w:9.25,h:0.48,fontSize:9.5,bold:true,color:C.purple,align:"center",valign:"middle"});
}

// ── SLIDE 6: H&E STAIN ───────────────────────────────
{
  const s=pres.addSlide();
  bg(s,C.off);
  hdr(s,"Haematoxylin & Eosin (H&E) Stain — Procedure & Results",C.navy);

  s.addShape(pres.shapes.RECTANGLE,{x:0.28,y:1.05,w:9.45,h:0.38,fill:{color:"EBF4FF"},line:{color:C.navy,width:0.5}});
  s.addText("PRINCIPLE: Haematoxylin (basic dye) stains acidic components (DNA/RNA in nucleus) BLUE. Eosin Y (acidic dye) stains basic components (cytoplasm, collagen, RBCs) PINK/RED. Used for BOTH histological tissue sections AND cytological smears.",
    {x:0.36,y:1.05,w:9.28,h:0.38,fontSize:9.5,color:C.text,valign:"middle"});

  colHdr(s,0.28,1.48,5.4,0.32,"H&E PROCEDURE FOR CYTOLOGICAL SMEAR",C.fix);
  const heSteps=[
    "Fix in 95% ETHANOL — 15 minutes (wet fixation)",
    "Rinse with distilled water",
    "HARRIS HEMATOXYLIN — 3-5 minutes",
    "Water rinse — 2 minutes",
    "Acid-alcohol differentiation (1% HCl) — few dips",
    "Water rinse; SCOTT'S TAP WATER or ammonia (bluing) — 1-2 min",
    "Water rinse; 70% → 95% ethanol (dehydration)",
    "EOSIN Y (1% aqueous or alcoholic) — 2-3 minutes",
    "95% → 100% ethanol (×2) — dehydration",
    "XYLENE (×2) — clearing; mount with DPX",
  ];
  heSteps.forEach((st,i)=>stepDot(s,i+1,st,0.28,1.83+i*0.38,5.4,C.fix));

  colHdr(s,5.85,1.48,3.88,0.32,"RESULTS (H&E)",C.fix);
  const heRes=[["Nuclei","Blue / dark purple (haematoxylin)"],["Cytoplasm","Pink / red (eosin)"],["RBCs","Bright red/orange"],["Mucus","Pale pink"],["Keratin","Deep pink"],
    ["Collagen fibers","Pink"],["Muscle","Deep pink (darker than collagen)"],["Malignant cells","Hyperchromatic nuclei, increased N:C"]];
  heRes.forEach((r,i)=>{
    const y=1.83+i*0.41;
    s.addShape(pres.shapes.RECTANGLE,{x:5.85,y,w:3.88,h:0.39,fill:{color:i%2===0?C.off:C.white},line:{color:C.silver,width:0.5}});
    s.addText(r[0],{x:5.92,y:y+0.01,w:3.72,h:0.18,fontSize:8.5,bold:true,color:C.fix});
    s.addText("→ "+r[1],{x:5.92,y:y+0.2,w:3.72,h:0.16,fontSize:8,color:C.text,italic:true});
  });
  s.addShape(pres.shapes.ROUNDED_RECTANGLE,{x:0.28,y:5.0,w:9.45,h:0.48,fill:{color:"EBF4FF"},line:{color:C.fix,width:0.75},rectRadius:0.08});
  s.addText("H&E used for BOTH histological sections (tissue) AND cytological smears | Nuclei = BLUE | Cytoplasm = PINK — simple 2-color rule",
    {x:0.38,y:5.0,w:9.25,h:0.48,fontSize:10,bold:true,color:C.fix,align:"center",valign:"middle"});
}

// ── SLIDE 7: GI TRACT ────────────────────────────────
{
  const s=pres.addSlide();
  bg(s,C.off);
  hdr(s,"Gastrointestinal Tract — Specimens & Smear Preparation",C.gi);

  colHdr(s,0.28,1.05,4.55,0.32,"GI TRACT SPECIMENS (Cytology)",C.gi);
  tbl(s,
    ["Specimen","Source","Purpose"],
    [
      ["Gastric washings","Stomach via endoscope/NG tube","Gastric carcinoma, H. pylori"],
      ["Oesophageal brushings","Oesophagus via endoscope","Carcinoma, Barrett's oesophagus"],
      ["Colonic/rectal brushing","Colonoscope","Colorectal carcinoma"],
      ["Ascitic fluid","Peritoneal cavity (aspiration)","Peritoneal metastases"],
      ["Liver FNAC","US-guided needle","Liver malignancy"],
    ],
    0.28,1.4,4.55,[1.6,1.6,1.35]
  );

  colHdr(s,5.0,1.05,4.73,0.32,"SMEAR PREPARATION — GI SPECIMENS",C.gi);
  const giSteps=[
    "GASTRIC WASHING / FLUID: Centrifuge 1500-2000 rpm × 10 min",
    "Discard supernatant; use the SEDIMENT",
    "Prepare 2-3 smears from sediment — spread thinly",
    "Fix IMMEDIATELY in 95% ethanol (Pap) OR air-dry (MGG)",
    "BRUSHING specimens: Roll brush directly onto glass slides",
    "Fix or air-dry IMMEDIATELY — cells transfer off brush rapidly",
    "Cell block: Centrifuge → clot with plasma-thrombin → formalin fix → process like tissue → H&E section",
  ];
  giSteps.forEach((st,i)=>stepDot(s,i+1,st,5.0,1.4+i*0.53,4.73,C.gi));

  // Key findings card
  s.addShape(pres.shapes.RECTANGLE,{x:0.28,y:4.42,w:4.55,h:1.05,fill:{color:"E8F5EE"},line:{color:C.gi,width:0.75}});
  s.addText("KEY CYTOLOGICAL FINDINGS IN GI:\n• Normal: Honeycomb pattern of gastric epithelial cells\n• Malignancy: ↑ N:C ratio, hyperchromasia, irregular nuclear membrane, prominent nucleoli\n• H. pylori: Curved rods on Giemsa stain (background)",
    {x:0.36,y:4.42,w:4.38,h:1.05,fontSize:9,color:C.gi,fontFace:"Calibri",valign:"top"});
}

// ── SLIDE 8: URINARY TRACT ───────────────────────────
{
  const s=pres.addSlide();
  bg(s,C.off);
  hdr(s,"Urinary Tract — Specimens & Smear Preparation",C.urine);

  colHdr(s,0.28,1.05,4.55,0.32,"URINARY TRACT SPECIMENS (Cytology)",C.urine);
  tbl(s,
    ["Specimen","Collection","Purpose"],
    [
      ["Voided urine","Clean catch, 2nd morning void","TCC screening, infection"],
      ["Bladder washing","Catheter/cystoscope instil+aspirate saline","Higher cellularity; TCC"],
      ["Ureteric specimen","Ureteric catheterization","Upper tract lesions"],
      ["Prostatic massage","Post-massage urine/secretion","Prostate carcinoma"],
      ["Renal pelvis brushing","Ureteroscopy","Renal pelvic carcinoma"],
    ],
    0.28,1.4,4.55,[1.6,1.65,1.3]
  );

  colHdr(s,5.0,1.05,4.73,0.32,"URINE SMEAR PREPARATION",C.urine);
  const uSteps=[
    "Collect 2nd MORNING VOID (NOT first — overnight cells degenerate)",
    "CENTRIFUGE 1500-2000 rpm × 10 minutes",
    "Discard supernatant; leave ~0.5-1 mL; resuspend sediment",
    "Transfer drops to glass slides; SPREAD THINLY",
    "Fix IMMEDIATELY in 95% ETHANOL (urine cells degenerate very rapidly!)",
    "OR use CYTOCENTRIFUGE (cytospin) — monolayer — PREFERRED for urine",
    "Stain with PAP (routine gold standard), H&E, or MGG",
  ];
  uSteps.forEach((st,i)=>stepDot(s,i+1,st,5.0,1.4+i*0.535,4.73,C.urine));

  s.addShape(pres.shapes.RECTANGLE,{x:0.28,y:4.42,w:4.55,h:1.05,fill:{color:"F3EEF8"},line:{color:C.urine,width:0.75}});
  s.addText("KEY CYTOLOGICAL FINDINGS IN URINE:\n• Normal: Umbrella cells (large superficial transitional cells)\n• Malignancy (TCC): ↑ N:C ratio, hyperchromasia, irregular nuclear membrane\n• Decoy cells: BK polyomavirus / CMV (nuclear viral inclusions)\n• Inflammatory cells → Cystitis / UTI",
    {x:0.36,y:4.42,w:4.38,h:1.05,fontSize:9,color:C.urine,fontFace:"Calibri",valign:"top"});
  s.addShape(pres.shapes.RECTANGLE,{x:5.0,y:5.25,w:4.73,h:0.28,fill:{color:"F3EEF8"},line:{color:C.urine,width:0.75}});
  s.addText("⚠  PAP stain is GOLD STANDARD for urine cytology — excellent nuclear detail + transparent cytoplasm",
    {x:5.08,y:5.25,w:4.58,h:0.28,fontSize:9,color:C.urine,bold:true,valign:"middle"});
}

// ── SLIDE 9: THREE STAINS COMPARISON ─────────────────
{
  const s=pres.addSlide();
  bg(s,C.off);
  hdr(s,"Pap vs. MGG vs. H&E — Comparison Table (RGUHS Exam)",C.navy);

  tbl(s,
    ["Feature","PAP STAIN","MGG STAIN","H&E STAIN"],
    [
      ["Fixation","Wet — 95% ethanol IMMEDIATE","AIR DRYING (before staining)","Wet — 95% ethanol"],
      ["Nuclear detail","★★★★★ EXCELLENT","★★★ Good","★★★★ Very good"],
      ["Cytoplasmic detail","★★★★ Very good","★★★★★ EXCELLENT","★★★ Moderate"],
      ["Transparency","Transparent (overlapping cells readable)","Coloured background","Moderate"],
      ["Best for","All tracts, gynae smears, malignancy","FNAC, lymphoma, haematology","Histology sections + cytology"],
      ["Nuclei colour","BLUE-BLACK (hematoxylin)","PURPLE-BLUE/VIOLET","BLUE/DARK PURPLE"],
      ["Cytoplasm colour","GREEN (EA) / ORANGE (OG-6) / PINK","BLUE (basophilic) / PINK (eosinophilic)","PINK (eosin Y)"],
      ["Keratin","ORANGE (OG-6)","Pink-red","Deep pink"],
      ["Gold standard","Urine + Gynaecological cytology","FNAC smears","Histopathology"],
      ["Staining duration","~1.5-2 hours (multiple steps)","~25-30 min (faster)","~45 min"],
    ],
    0.25,1.12,9.5,[2.0,2.5,2.5,2.5]
  );

  s.addShape(pres.shapes.ROUNDED_RECTANGLE,{x:0.25,y:4.7,w:9.5,h:0.72,fill:{color:C.navy},line:{color:C.navy,width:0},rectRadius:0.1});
  s.addText("KEY RULE: WET FIX → Pap & H&E | AIR DRY → MGG\nPap = BEST nuclear detail | MGG = BEST cytoplasm/background | H&E = Tissue AND cytology (dual purpose)",
    {x:0.35,y:4.7,w:9.3,h:0.72,fontSize:11.5,bold:true,color:C.yellow,align:"center",valign:"middle"});
}

// ── SLIDE 10: CYTOLOGICAL FINDINGS ALL 3 TRACTS ───────
{
  const s=pres.addSlide();
  bg(s,C.off);
  hdr(s,"Cytological Findings — All Three Tracts (What to See in Exam)",C.navy);

  const cols=[
    {title:"RESPIRATORY TRACT",col:C.resp,x:0.28,w:3.1,items:[
      ["Normal cells","Bronchial columnar cells with cilia; mucous cells; alveolar macrophages"],
      ["Squamous metaplasia","Polygonal cells with dense cytoplasm — precancerous change"],
      ["Squamous cell carcinoma","Abnormal squamous cells; keratin pearls; tadpole/fibre cells"],
      ["Adenocarcinoma","Glandular cells; acinar/papillary pattern; mucin vacuoles"],
      ["Malignant features","↑ N:C ratio; hyperchromasia; irregular nuclear membrane; prominent nucleoli"],
      ["Pulmonary macrophages","Phagocytic cells with ingested particles; seen in smokers (carbon)"],
    ]},
    {title:"GASTROINTESTINAL TRACT",col:C.gi,x:3.52,w:3.1,items:[
      ["Normal gastric cells","Honeycomb pattern; columnar with basal nucleus"],
      ["H. pylori","Curved rods on Giemsa; on surface of epithelial cells"],
      ["Gastric carcinoma","Single cells or clusters; high N:C ratio; signet ring cells (mucin vacuole pushing nucleus)"],
      ["Colorectal carcinoma","Columnar cells; dirty necrotic background (Ziegler's necrosis)"],
      ["Normal colonic cells","Tall columnar cells with goblet cells"],
      ["Inflammatory cells","Neutrophils, lymphocytes in ulcerative colitis/infection"],
    ]},
    {title:"URINARY TRACT",col:C.urine,x:6.76,w:3.0,items:[
      ["Umbrella cells","Large superficial transitional cells with eosinophilic cytoplasm — NORMAL"],
      ["TCC (low grade)","Mild nuclear enlargement; slight hyperchromasia"],
      ["TCC (high grade)","Marked ↑ N:C; irregular nuclear contour; coarse chromatin; prominent nucleoli"],
      ["Decoy cells (BK virus)","Smudgy 'owl-eye' nuclear inclusion — BK polyomavirus"],
      ["CMV cytopathic effect","Large cell; intranuclear 'owl-eye' inclusion; cytoplasmic inclusions"],
      ["Squamous cells","Contamination from vaginal flora (females)"],
    ]},
  ];

  cols.forEach(col=>{
    s.addShape(pres.shapes.RECTANGLE,{x:col.x,y:1.05,w:col.w,h:0.32,fill:{color:col.col},line:{color:col.col,width:0}});
    s.addText(col.title,{x:col.x,y:1.05,w:col.w,h:0.32,fontSize:8.5,bold:true,color:C.white,align:"center",valign:"middle",margin:0});
    col.items.forEach((it,i)=>{
      const y=1.4+i*0.69;
      s.addShape(pres.shapes.RECTANGLE,{x:col.x,y,w:col.w,h:0.67,fill:{color:i%2===0?C.off:C.white},line:{color:C.silver,width:0.5}});
      s.addText(it[0],{x:col.x+0.06,y:y+0.02,w:col.w-0.1,h:0.22,fontSize:9,bold:true,color:col.col});
      s.addText(it[1],{x:col.x+0.06,y:y+0.24,w:col.w-0.1,h:0.38,fontSize:8.5,color:C.text,valign:"top"});
    });
  });
}

// ── SLIDE 11: EXAM QUESTIONS + MEMORY AIDS ────────────
{
  const s=pres.addSlide();
  bg(s,C.off);
  hdr(s,"RGUHS Exam Questions & Memory Aids — Pathology",C.navy);

  colHdr(s,0.28,1.05,5.3,0.32,"RGUHS EXAM QUESTIONS — PATHOLOGY",C.navy);
  s.addShape(pres.shapes.RECTANGLE,{x:0.28,y:1.4,w:5.3,h:3.78,fill:{color:C.white},line:{color:C.silver,width:0.75},shadow:{type:"outer",color:"000000",blur:5,offset:2,angle:135,opacity:0.08}});

  s.addText("LONG ANSWER (10 marks):",{x:0.38,y:1.46,w:5.1,h:0.28,fontSize:10,bold:true,color:C.resp});
  const lq=["• Describe smear preparation and Pap staining of sputum specimen — principle, procedure, results","• Describe collection, smear preparation and staining of urine for cytological examination","• Compare Papanicolaou stain with MGG stain under — principle, fixation, procedure, results","• Processing of specimens in cytology laboratory — GI, respiratory, urinary tract"];
  lq.forEach((q,i)=>s.addText(q,{x:0.38,y:1.76+i*0.35,w:5.1,h:0.33,fontSize:9.5,color:C.text}));

  s.addText("SHORT ANSWER (5 marks):",{x:0.38,y:3.22,w:5.1,h:0.28,fontSize:10,bold:true,color:C.resp});
  const sq=["• Fixatives used in cytology — types and uses","• Wet vs. dry fixation — differences and uses","• Cytocentrifuge (cytospin) — principle, procedure, advantages","• Urinary sediment cytology — collection and processing","• Normal cells in Pap-stained sputum smear"];
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  s.addText("VSA (2 marks):",{x:0.38,y:5.17,w:5.1,h:0.24,fontSize:10,bold:true,color:C.resp});
  s.addText("• 3 components of EA stain  • Decoy cells in urine  • Difference wet/dry fixation  • 2 respiratory cytology specimens",
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  const aids=[
    {t:"Pap Reagents","b":"H-A-B-OG-EA\n(Haem → Acid-alcohol → Bluing → OG-6 → Eosin Azure)"},
    {t:"Pap Colours: \"BOP\"","b":"B=Black nuclei | O=Orange superficial | P=Pink cytoplasm"},
    {t:"Wet Fix = Pap & H&E","b":"Fix BEFORE drying — 95% ethanol IMMEDIATELY"},
    {t:"Dry Fix = MGG","b":"Air dry COMPLETELY first — then methanol fix"},
    {t:"Best nuclear detail","b":"Pap > H&E > MGG"},
    {t:"Best cytoplasm/background","b":"MGG > Pap > H&E"},
  ];
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// ── SLIDE 12: KEY TAKEAWAYS ───────────────────────────
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  s.addText("KEY POINTS TO REMEMBER",{x:0.5,y:0.2,w:9,h:0.62,fontSize:26,bold:true,color:C.teal,align:"center",charSpacing:4});

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    {n:"1",t:"FIXATION is the first and most critical step — wrong fixation = ruined smear. Pap/H&E = WET (alcohol). MGG = AIR DRY."},
    {n:"2",t:"PAP STAIN — best nuclear detail; gold standard for urine and respiratory cytology. 5 dyes in 3 solutions. Sequence: H-A-B-OG-EA."},
    {n:"3",t:"MGG STAIN — best for cytoplasm and background material; excellent for FNAC, lymphoma. Air-dry smear BEFORE staining."},
    {n:"4",t:"H&E — Nuclei = BLUE, Cytoplasm = PINK. Used for both tissue sections and cytological smears."},
    {n:"5",t:"RESPIRATORY: Select purulent portion; fix immediately; cytocentrifuge for washings/BAL. Bronchial brushings smeared directly."},
    {n:"6",t:"URINARY: 2nd morning void; centrifuge; fix IMMEDIATELY (cells degenerate fast). Decoy cells = BK virus. Umbrella cells = normal urothelium."},
  ];
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  s.addText("3rd B.Sc. MLT  |  RGUHS Pathology  |  Cytopathology — Smear Preparation & Staining",
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// ── WRITE ─────────────────────────────────────────────
pres.writeFile({fileName:"/home/daytona/workspace/rguhs-path-ppt/RGUHS_Pathology_Smear_Staining_3rdMLT.pptx"})
  .then(()=>console.log("✅ Done!"))
  .catch(e=>console.error("❌",e));

cd /home/daytona/workspace/rguhs-path-ppt && node create_path_ppt.js

ls -lh /home/daytona/workspace/rguhs-path-ppt/

cd /home/daytona/workspace/rguhs-path-ppt && libreoffice --headless --convert-to…

cp /home/daytona/workspace/rguhs-path-ppt/RGUHS_Pathology_Smear_Staining_3rdMLT.…

mv /home/daytona/Smear_Staining_Pathology.pdf /home/daytona/workspace/rguhs-path…