### **SEMEN ANALYSIS** ## Definition * Semen = **spermatozoa (sperms) + seminal fluid** * ~40% infertility cases due to semen abnormalities * First investigation in infertility workup * Defects: * **Quantitative** → absent/low sperm number * **Qualitative** → abnormal sperm function/morphology --- ## Indications * Assessment of fertility/infertility * Monitor success of: * Varicocelectomy * Vasectomy * Assess donor semen for artificial insemination * Medicolegal purpose: * Alleged rape cases → vaginal pool smear for sperm detection * Selection of assisted reproductive techniques: * IVF * GIFT --- # Collection of Sample ### Requirements * Masturbation collection * Ejaculatory abstinence: **2–7 days** * Collect in: * Clean * Dry * Wide-mouthed plastic/glass container ### Avoid Condom Sample * May contain spermicidal agents * Impair sperm motility --- # Examination of Semen # I. Physical Examination ## 1. Liquefaction ### Normal * Immediately after ejaculation → semisolid coagulated mass * Begins liquefying within few minutes * Complete liquefaction within **15 min** * By **30 min** → homogeneous, watery ### Abnormal * Failure to liquefy → inadequate prostate secretion --- ## 2. Semen Viscosity ### Method * Aspirate semen into wide-bore pipette (~1.5 mm) * Allow drop by gravity ### Normal * Falls drop by drop ### Increased Viscosity * Thread >2 cm long ### Significance * Increased viscosity ↓ sperm motility --- ## 3. Appearance ### Normal * Fresh semen: * Opaque * White-gray * Viscid ### After Liquefaction * Homogeneous gray-opalescent ### Abnormal Colors | Color | Cause | | ------------ | --------------------------------- | | Red-brown | RBCs (hemospermia) | | Yellow | Jaundice, certain vitamins/drugs | | Yellow hue | Pyospermia | | Rust colored | Small bleeding in seminal vesicle | --- ## 4. Semen Volume ### Normal * > 1.4 mL ### Measurement * Weigh sample * Density assumed = **1 g/mL** ### Low Volume * Ejaculatory duct obstruction * Congenital bilateral absence of vas deferens * Collection difficulty ### High Volume * Active exudation due to inflammatory lesions of accessory organs --- ## 5. Semen pH ### Normal * Alkaline * **7.2–8** * > 7.2 ### Measurement * After liquefaction * Preferably after 30 min --- # II. Microscopic Examination ## 1. Sperm Aggregation vs Agglutination ### Aggregation * Nonspecific * Immotile sperms stick together * Motile sperms attach to: * Mucus strands * Debris * Nonsperm cells ### Agglutination * Motile sperms stick to each other: * Head-to-head * Tail-to-tail * Mixed pattern --- ## 2. Cellular Elements Other Than Sperms Look for: * Epithelial cells from genitourinary tract * Round cells: * Leukocytes * Immature germ cells --- ## 3. Sperm Motility ### Importance * Cervical mucus penetration * Migration into fallopian tube ### Normal * **42% (40–43%) motile sperms** Includes: * Rapidly progressive * Slowly progressive ### Method * Drop of liquefied semen on slide * Coverslip * Microscopic examination ### WHO 2021 Categories #### Rapidly Progressive * 29–31% * Movement >5 head lengths/sec #### Slow Progressive * ~1% #### Immotile * 54% (50–56%) --- ## 4. Sperm Vitality ### Purpose * Determine whether immotile sperms alive or dead ### Normal * **54% (50–56%) live forms** ### Based On * Intact cell membrane ### Especially Important * Progressive motility <40% ### Methods * Dye exclusion test * Hypotonic swelling test --- ## 5. Total Sperm Count ### Method * Improved Neubauer chamber * Thoma pipette * Dilution 1:20 * Same as TLC method ### Normal * > 39 million/ejaculate ### Terms #### Aspermia * No semen #### Azoospermia * No spermatozoa in ejaculate #### Oligospermia * Sperm count below lower reference limit --- # 6. Sperm Morphology ### Stain * Papanicolaou stain ### Normal Sperm Parts * Head * Neck * Midpiece * Principal piece * Endpiece ### Practical View * Head + neck * Tail (midpiece + principal piece) ### Normal Morphology * > 4% normal forms ### Requirement * Both head and tail normal ### Otherwise * Abnormal sperm --- # Classification of Abnormal Sperm Morphology ## A. Head Defects ### Abnormal Size * Large head * Small head ### Abnormal Shape * Pointed * Tapered * Round * Pyriform * Amorphous ### Acrosomal Defects * Vacuolated acrosomal region * Abnormal acrosomal area ### Abnormal Number * Double-headed sperm --- ## B. Neck & Midpiece Defects * Sharply bent neck * Asymmetrical midpiece insertion * Thick midpiece * Thin midpiece * Irregular midpiece --- ## C. Tail (Principal Piece) Defects * Short tail * Bent tail * Coiled tail * Double tail --- # Tests for Antibody Coating of Spermatozoa ## Antisperm Antibodies (ASA) ### Cause * May produce sperm agglutination ### Types * IgA * IgG ### Present In * Semen * Seminal plasma * Blood serum * Cervical mucus --- ## Direct Tests (Detect antibodies on sperm) ### 1. MAR Test * Mixed Antiglobulin Reaction test ### 2. Immunobead (IB) Test --- ## Indirect Tests (Detect antisperm antibodies in sperm-free fluids) * Performed on: * Seminal plasma * Serum * Cervical mucus --- ### Ultra-Short Revision Box **Normal Values** * Volume → **>1.4 mL** * pH → **7.2–8** * Motility → **42%** * Vitality → **54%** * Sperm count → **>39 million/ejaculate** * Normal morphology → **>4%** **Definitions** * Aspermia = No semen * Azoospermia = No sperms * Oligospermia = Low sperm count * Hemospermia = Blood in semen * Pyospermia = Pus cells in semen * Agglutination = Motile sperms stick to each other " create masterchart of semen analysis from this text, including everything. In photo format and in just one page.

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