Clin Path Lab (Dra.

Sionzon)

Seminal Fluid Analysis & Fertility Assessment

10 December 07

SEMINAL FLUID ANALYSIS & FERTILITY ASSESSMENT

…APPEARANCE…
PHYSIOLOGY • color may be altered by length of abstinence
• the shorter the duration, the more transparent the
semen: 4 fractions sample is

SOURCE VOLUME CHARACTERISTICS APPEARANCE: gray to white opalescent

bulbourethral glands 0.1-0.2 cc viscous, clear
brown to reddish (+) blood
testes, epididymis 0.1-0.2 cc (+) sperm yellow certain drugs
prostate 0.5-1.0 cc acidic, watery contamination with urine
seminal vesicles 1.0-3.0 cc gelatinous, (+) prolonged abstinence
fructose turbid increased leukocytes --- infection in
COMPLETE 2.0-5.0 cc reproductive tract
EJACULATE:
clear low sperm concentration
spermatozoa --- seminiferous tubules
…LIQUEFACTION TIME…
• may contain jelly-like grains that do not liquefy
• may add bromelin --- for samples that do not liquefy
mature & stored in epididymis
LIQUEFACTION TIME: 10-30 mins
60% seminal vesicles  (+) high fructose content prolonged: >60 mins may contain mucus ---
---important for sperm infection
motility functional disturbance of the
 secretes protein kinase prostate gland
responsible for coagulation
20- prostate gland  acidic
30%  ACP, citric acid, Zn, VISCOSITY: pours in droplets
proteolytic enzymes --- string or thread-like increased viscosity --- interfere
important in liquefaction of more than 2 cm with sperm mobility &
5% bulbourethral  thick & alkaline fertilizing ability
glands  neutralize acidity from prostate dysfunction 2° to
prostate & vagina chronic inflammation
5% spermatozoa & Rated 0 (watery) to 4 (gel-like)
fluid from
epididymis ..VOLUME…
SPECIMEN COLLECTION • clean graduated cylinder calibrated 0.1 ml increments
 majority of the sperm --- 1st portion ejaculate --- sample is aspirated using a wide-mouthed pipette
 abstinence: at least 3 days & not longer than 5 days • estimated by weighing the sample in the collection
 2-3 samples tested at 2-week intervals container --- 1 ml=1 gm
 warm, sterile glass/plastic containers VOLUME: 2-5 ml
 specimen collected at RT & delivered to the lab within 1 increased extended abstinence
hour --- time of collection varicocoele
 masturbation decreased infertility
 non-lubricant containing silicone condoms ·dysfunction of semen-
 fresh: clotted --- liquefies within 30-60 min producing organsincomplete
 analysis should not begin until after liquefaction specimen collection
has occurred
 awaiting analysis --- 37°C
…pH…
 biohazard
PARAMETERS: NORMAL VALUES:
appearance gray to white opalescent
liquefaction time 10-30 mins
viscosity pours in droplets
volume >2.0 ml
pH 7.2-7.8 (8.0)
sperm >20x106/ml
concentration
ANALYSIS PROPER total sperm count >40x106/ejaculate
motility >50% with forward progression
>25% with rapid progression
Brim, leu, virns 1 of 3
morphology >30% with normal morphology
(strict criteria)
>50% with normal morphology
(routine criteria)
WBC <1x106/ml
 Clin Path Lab – Seminal Fluid Analysis and Fertility Assessment by Dra Sionzon Page 2 of 3

• specimens not tested within 1 hour have pH changes 6 infertile

<5x10 /ml
• urinalysis reagent strip --- pH range 6.1-10.0
(-) azoospermia
no ejaculate aspermia
pH: 7.2-7.8 (8.0)
decreased increased prostatic fluid
…MORPHOLOGY…
increased reproductive tract infection
• thinly smeared --- feathering technique
extremely acidic agenesis or occlusion of the
seminal vesicles • Papanicolao stain --- most widely used
extremely decreased prostatic secretion • Giemsa, Wright’s, Harris Hematoxylin
alkaline • examined under oil immersion --- 200 sperms
• air-dried slides are stable for 24 hours
…MOTILITY… • information on the function of the reproductive tract &
• forward progression describes how fast the motile predictor of fertility
sperm are moving • physical aberration --- production or during storage in
• poor motility --- sperm will not travel through the the epididymis
cervical mucosa, uterus, then the fallopian tubes to
reach ovum MORPHOLOGY: >30% with normal morphology
• high percentage of immotile sperm & clumping requires (strict criteria)
further evaluation >50% with normal morphology
• 10 uL, 22x22 mm coverslip, 20hpf (routine criteria)
 oval-shaped head, 5x3 um
MOTILITY: >50% with forward progression  long, flagellar tail, 45 um
>25% with rapid progression  unremarkable neck & middle piece
<50% after 2 hrs asthenozoospermia  acrosomal cap encompassing ½ of the head
infertility
>50% abnormal any single abnormality or
graded as follows: morphology combination--- spermatozoa
4.0 rapid, straight-line motility may not penetrate ovum
head: large/small, tapering, pyriform,
amorphous, vacuolated, double
3.0 slower speed, some lateral
movement neck & midpiece: absent/non-inserted/bent tail;
distended, irregular, bent, thin
2.0 slow forward progression, tail: short, multiple, hairpin, broken,
noticeable lateral movement irregular width, coiled, w/
terminal droplets
1.0 no forward progression cytoplasmic droplets: greater than 1/3 of the head

0 no movement

..SPERM CONCENTRATION/COUNT..
• diluent: 100 ml dist. H2O + 1 ml neutral buffered
formalin + 5 gm NaHCO3
• 1:20 dilution --- 0.5 ml of semen + 10 ml diluent
thoroughly mixed
• charge 2 chambers Neubauer hemocytometer & count
after 2 mins --- average the 2 counts
• use R squares
• may add stain --- crystal violet
• only fully developed sperms are counted
• “round cells” are counted separately

 # of sperms x 1M = sperms/ml
 # of sperms/ml x volume of ejaculate = teratospermia syndromes globospermia
sperms/ejaculate (total sperm count) microcephaly
short tail spermatozoa
increased number of immature epididymal dysfunction
SPERM CONCENTRATION: 6 forms frequent ejaculations
>20x10 /ml
increased number of tapering heads varicocoele
TOTAL SPERM COUNT: 6
>40x10 /ejaculate ·spermatozoa with cytoplasmic
6 oligospermia dropletsspermatozoa with bent tail
<20x10 /ml sub-fertile
Clin Path Lab – Seminal Fluid Analysis and Fertility Assessment by Dra Sionzon Page 3 of 3