Spermanalysis Flashcards
Stages of sperm maturation
Spermatogonium, 1st spermatocyte, 2nd spermatocyte, spermatid, spermatozoon
Composition of semen 5%
Spermatozoa
Composition of semen 60-70%
Seminal fluid
Composition of semen 20-30%
Prostate fluid
Composition of semen 5%
Bulbourethral gland fluid
Site of spermatogenesis
Seminiferous tubules (testes)
Nurse cells for developing sperm
Sertoli cells
Site of sperm maturation
Epididymis
Propels sperm to ejaculatory ducts
Vas deferens (ductus deferens)
Spermatogenesis and sperm maturation time
90 days (74 days in Graff cycle)
Acidic fluid containing ACP, zinc, citric acid, and enzymes
Prostate fluid
Bulbourethral gland secretes alkaline mucus for
Neutralizing acidity from prostatic secretions and vagina
Specimen collection for sperm abstinence
2-7 days
Prolonged abstinence effect
Increased volume, decreased motility
WHO recommendation for specimen collection
Two to three specimens, not less than 7 days or more
First portion of sperm missing effect
Decreased sperm count, increased pH, specimen will not liquefy
Last portion of sperm missing effect
Increased sperm count, decreased pH and volume, specimen will not clot
Best method for sperm analysis
Masturbation
Condom requirements for specimen collection
Non-lubricant-containing rubber or polyurethane condom
Specimen for sperm analysis should be delivered within
1 hour of collection at room temperature
Liquefaction for sperm analysis should be done
30-60 minutes
Failure to liquefy sperm within 60 minutes causes
Deficiency in prostatic enzymes
Failure to liquefy sperm remedy
Treat with amylase, bromelain, α-chymotrypsin, or DPBS to break up mucus
Specimen awaiting analysis should be kept at
37°C (water bath)
Normal appearance of sperm
Gray-white, translucent (due to flavin)
Odor of sperm
Musty or bleach odor
Increased white turbidity in sperm indicates
Infection
Red or brown coloration in sperm indicates
Increased blood or RBCs
Yellow coloration of sperm indicates
Urine contamination, medication, abstinence
Normal volume of sperm
2-5 mL
Decreased volume of sperm indicates
Infertility, incomplete collection
Normal viscosity of sperm
Pours in droplets
Abnormal viscosity of sperm
Threads >2 cm long
Higher viscosity effect on motility
Decreased motility
Reporting of sperm
0 - watery, 4 - gel-like
pH of normal sperm
7.2 to 8.0
Increased pH in sperm indicates
Infection
Decreased pH in sperm indicates
Increased prostatic fluid
Normal value of sperm concentration
> 20 million/mL
Dilution of sperm in improved Neubauer counting chamber
1:20
Diluents for sperm purpose
Immobilize sperm
Examples of diluents for sperm
Formalin, sodium bicarbonate (NaHCO3), saline, distilled water, cold tap water
Shortcut method for sperm count computation (2 WBCs square)
sperm counted x 100,000
Shortcut method for sperm count computation (5 RBCs square)
sperm counted x 1,000,000
Sperm count normal value
> /= 40 million/ejaculate
Sperm motility normal value
50% motility within 1 hour
Quality of sperm motility should be
> /= 2.0
Grading of sperm (WHO criteria: rapid straight motility)
4 (a)
Grading of sperm (WHO criteria: slower speed, some lateral movement)
3.0 (b)
Grading of sperm (WHO criteria: slow forward progression, noticeable lateral movement)
2.0 (b)
Grading of sperm (WHO criteria: no forward progression)
1.0 (c)
Grading of sperm (WHO criteria: no movement)
0 (d)
Alternative sperm motility grading criteria: sperm moving linearly in a large circle
Progressive motility
Alternative sperm motility grading criteria: sperm moving with absence of progression
Nonprogressive motility
Alternative sperm motility grading criteria: no movement
Immotility
Determines sperm concentration, morphology, velocity, and trajectory
Computer Assisted Semen Analysis (CASA)
Sperm morphology routine criteria normal forms
> 30%
Sperm morphology Kruger’s strict criteria
> 14%
Preferred stain for sperm morphology
Papanicolaou’s stain
Other stains for sperm morphology
Wright’s stain, Giemsa stain, Shorr stain
Contains mitochondria in sperm
Midpiece
Acrosomal cap measurement to head
1/2 of the head
Acrosomal cap measurement to nucleus
2/3 of the nucleus
Acrosomal cap measurement
3 µm
Sperm head size
5 µm
Neck of sperm size
7 µm
Tail measurement including neck
45 µm
Seminal fluid fructose is measured within
2 hours or frozen to prevent glycolysis
Screening test for seminal fructose
Resorcinol (Seliwanoff’s test)
Color for Seliwanoff’s test
Orange-red color
Cause of sperm agglutination, detected in semen, cervical mucosa, or serum
Antisperm antibodies
Detects the presence of IgG antibodies
Mixed agglutination reaction
Immunobead test detects the presence of
IgG, IgM, IgA antibodies
Presence of beads on sperm (normal)
<50%
Round cells in sperm
WBCs or spermatids (immature sperm cells)
WBCs and immature sperm cells are differentiated using
Peroxidase
Round cell count formula
NxS / 100
Detection of semen more specific method
Glycoprotein p30 (PSA)
Test for choline medicolegal
Florence test
Reagents for Florence test
Iodine crystals + Potassium iodide
Positive reaction in Florence test
Dark brown rhombic crystals
Barbiero’s test is used to detect
Spermine
Reagents for Barbiero’s test
Saturated picric acid + TCA
Positive reaction in Barbiero’s test
Yellow leaf-like crystals
Sperm count following vasectomy should ideally be
Zero within 12 weeks after procedure
Post vasectomy semen analysis is done after
2 months or 8 weeks
Post vasectomy is collected
2 months until 2 consecutive monthly specimens show no sperm
Eosin nigrosin stain is used for
Decreased motility with normal count
Decreased sperm count test
Fructose level
Decreased motility with clumping cause
Male antisperm antibodies
MAR and immunobead test and sperm agglutination with sperm serum is used for
Male antisperm antibodies
Normal analysis with continued infertility test
Sperm agglutination with female serum
Observing sperm penetration ability of partner’s midyear cervical mucus
Cervical mucus penetration
Sperms exposed to low-sodium concentration are evaluated for
Hypo-osmotic swelling
Evaluation of the acrosome to produce enzyme essential for ovum penetration
In vitro acrosome reaction
No semen or ejaculate
Aspermia
Absence of spermatozoa in the ejaculate
Azoospermia
Presence of blood in the ejaculate
Hematospermia
Increased number of leukocytes in ejaculate
Leukospermia
Increased number of immotile or dead spermatozoa in the ejaculate
Necrozoospermia
Decreased sperm concentration
Oligozoospermia
Hormones that stimulate spermatogenesis
FSH and LH
Leydig cells secrete
Testosterone
Sertoli cells secrete
Inhibin
Specimen for sperm analysis according to WHO
2 or 3 samples from <7 days or >3 weeks apart with 2 abnormal samples are considered significant
As the sperm die off, what remains and may be present after 7 days of intercourse
Head