Semen Analysis
Semen analysis is a basic investigation for the male partner apart from the other investigations. With out basic semen analysis one should never start the treatment of the female partner.

Semen analysis is divided under following headings.

• Collection
• Gross examination
• Microscopic Examination: Counting, Motility, Morphology
• Biochemical Investigation
• Automation

HEAD Shape: Oval with regular outline Length: 4-5.5 microns Width: 2.5-3.5 microns MID PIECE Width: Less then 1/3 the width of  head  Length: 6-10 microns TAIL Shape: Slender uncoiled & with regular outline Length: 45 microns

1. Collection

• Instruction to the Patients
• Days of sexual abstinence
• Room Infrastructure
• Collect by masturbation 
• Splite Ejaculation
• Condom
• Labeling   
• Name
• Date
• Time
• Safe Handling

2. Gross Examination

• Coagulation and liquefaction
• Coagulation
• Liquefaction < 30 min.
• Viscosity
  • Put sample for 10min more >Gentle Pipetting >Add Media
• Volume: 
  • 0.7 to 5.0 ml
  • By Graduated Cylinder with conical base.
  • Aspermia                        
  • Oligospermia                            
  • Hyperspermia                          

3. Microscopic Examination

• Wet preparation
  • 10µl semen with 18mm cover-slip
  • Assessment of Motility & Count

                                           Count/HPF
          Approximate count = ----------------- million/ml
                                                 4

• Number of Sperms / hpf
• Centrifuge for 10 min at 1000rpm
• Discard the supernant.
• Make slide from the palate
• For exact Count –  Advice for dilution

• By Improved Neubauer’s  Chamber
• By Meckler’s chamber
• Normal Count
  • Normal sperm count is 20 -150 millions/ml.
• Oligozoospermia
  • If the sperm count is less than 20 millions/ml.
• Azoospermia
  • When there are no sperms in the semen sample or when the sperm count is nil.
• ASTHENOZOOSPERMIA        
• An absence or a marked reduction (< 50% )in sperm movement.
• OLIGOASTHENOZOOSPERMIA
  • Decreased sperm movement and Decreased count.
• Sperm Motility Grading

          Non motile       - No Movement
          Sluggish motile - Side to Side Movement
          Slow Movement / partly moving forwards
                                   < 50 micron per second
          Actively motile   - Good forward movement
                                 - Excellent rapid forward progression.

• Sperm Vitality
• Eosin–Nigrosin Stain

• Morphology
• Two smear from the fresh semen sample
• Wet and Incomplete Air Drying
• For Swim up / 95 % Ethanol / Albumin
• Fixation of Smear
• 95 % Ethanol for 15 min
• Spray Fixative
• Staining of Smear
• PAP  Stain        

• Abnormality in Morphology
  • Teratozoospermia
  • Teratoasthenozoospermia                     
  • Head abnormalities
  • Mid piece abnormalities
  • Tail abnormalities     
  • Major sperm defect related to infertility are
    • Pear shaped Head
    • Coiled tailed defect
    • Proximal Cytoplasmic droplets

4. Biochemical Examination

• Semen Fructose for  Azospermia
• Antisperm Antibody Test (IgG & IgA)
  • Immunobead Test.
  • MAR Test.

5. Automation

• SQA II C Sperm Quality Analyzer
• CASA Computer Aided Semen Analysis.