SEMINALYSIS

5% spermatozoa, 60–70% seminal vesicle fluid (nutrients, buffers), 20–30% prostatic fluid (enzymes, proteins), and 5% bulbourethral gland secretions (alkaline mucus)

What are the components and contributions to semen composition?

Collected via masturbation after 2–7 days of abstinence using sterile containers at 37°C. Must be submitted within 1 hour.

What are the proper methods and conditions for semen specimen collection?

special condoms (non-spermicidal)

Alternative for semen specimen collection

Coitus interruptus

unreliable due to possible loss of first, sperm-rich portion

Volume: ≥1.5 mL
pH: >7.2
Sperm concentration: ≥15 million/mL
Total count: ≥39 million/ejaculate
Motility: ≥40% total, ≥32% progressive
Morphology (strict): ≥4% normal
Vitality: ≥58% live
Round cells: <1 million/mL

What are the WHO reference values for semen analysis?

gray-white, translucent, and has a musty odor

how does semen normally appear

urine or abstinence

yellow semen suggests

RBCs

red semen suggests

infection (↑WBCs)

turbid semen suggests

Leukocyte esterase tests

help differentiate cells

prostatic enzymes

Normal semen clots and liquefies within 30–60 minutes due to

prostatic deficiency

Failure to liquefy suggests

Liquefaction

necessary before analysis

DPBS or proteolytic enzymes (e.g., bromelain)

Liquefaction is assisted using

Using hemocytometer (Neubauer), 1:20 dilution
Total Sperm Count = Sperm concentration × Volume

How is sperm concentration and total sperm count calculated?

Grade 4 (a)

rapid, straight motility

Grade 3 (b)

slower, straight motility

Grade 2 (b)

slow progression motility

Grade 1 (c)

no progression motility

Grade 0 (d)

immotile

50% total motility or ≥25% progressive within 1 hour

normal motility

Thin smear stained (Wright’s, Giemsa, etc.), 200 sperm examined.

How is sperm morphology evaluated

Kruger’s strict criteria

head (5x3 µm), acrosome (½ head), midpiece, tail

Normal sperm morphology

≥4% normal forms (strict). Abnormalities affect function (e.g., double tail = hypermotility)

Round cells

WBCs and immature sperm

infection or spermatogenesis issue

>1M/mL round cells

peroxidase stain

Round cells are identified using —- to differentiate granulocytes

Vitality (eosin-nigrosin)

≥50% live sperm

Fructose (resorcinol test)

checks seminal vesicle function

Antisperm antibodies

assessed by MAR or immunobead test

Microbial test

for Chlamydia, Ureaplasma, etc.

Chemical markers

α-glucosidase (epididymis), zinc/citric acid (prostate)

Eosin-nigrosin staining: live sperm exclude dye (bluish-white), dead absorb (red)

How is sperm vitality tested

flagellar issue

Vital + immotile

epididymal defect

non-viable eosin-nigrosin stain

obstruction, congenital absence of vas deferens, or androgen deficiency

low fructose cause

orange color

Positive resorcinol

> 13 umol/ejaculate

Reference for fructose test

antisperm antibodies

Develop from blood-testis barrier disruption

MAR Test

<10% bound motile sperm = normal

immunobead Test

identifies IgG, IgM, IgA binding on sperm head/tail

male source

Clumping in immunobead test

female source

agglutination with cervical mucosa in immunobead test

Epididymis

↓ α-glucosidase, L-carnitine, glycerophosphocholine

Prostate

↓ zinc (≥2.4 μmol), citric acid (≥52 μmol), acid phosphatase (≥200 units)

Checks for sperm presence (motile/non-motile).
After 2 months, monthly exams until 2 consecutive samples are sperm-free.
Use wet mount → centrifuge if negative.

How is semen analyzed post-vasectomy?

Hamster egg penetration

evalutes ovum penetration

cervical mucus penetration

evaluates natural fertilization potential

hypo-osmotic swelling

evaluates membrane integrity, viability

in vitro acrosome reaction

evaluates enzyme release for penetration