What are the four fractions of Semen and its corresponding concentration in normal semen?
Spermatozoa - 5%
Seminal Fluid - 60-70%
Prostate Fluid - 20-30%
Bulbourethral Glands - 5%
A. Urinary Bladder
B. Vas Deferens
C. Urethra
D. Penis
E. Glans Penis
F. Seminal Vesicle
G. Rectum
H. Ejaculatory Duct
I. Prostate Gland
J. Anus
K. Bulbourethral Gland
L. Epididymis
M. Testis
N. Scrotum
A. Ureter
B. Vas deferens
C. Prostate gland
D. Penis
E. Urinary bladder
F. Seminal Vesicle
G. Bulbourethral Gland
H. Epididymis
I. Scrotum
J. Urethra
Where is the seminiferous tubules located?
testes
Precursor of Spermatozoa
Germ cells
they provide support and nutrients for the germ cells as they undergo spermatogenesis
Specialized Sertoli cells
where the sperm develops the flagella
epididymis
storage of sperm before ejaculation
epididymis
meeting place of the sperm and seminal fluid
ejaculatory ducts
this fluid contains a high concentration of fructose that comprises most of the semen
seminal fluid
source of energy of the sperm to move the flagella
fructose
located below the bladder that surrounds the upper urethra and aids in propelling the sperm through the urethra by contractions during ejaculation
prostate gland
comprise the acidic portion of the semen
prostate fluid
chemicals present in the prostate fluid that is responsible for both the coagulation and liquefaction of the semen
acid phosphatase
citric acid
zinc
proteolytic enzymes
produces the thick alkaline mucus of the semen
bulbourethral gland
When is the best time to collect sperm sample
2-3 days of sexual abstinence but not longer than 5 days
what is the effect of prolonged abstinence in sperm collection?
increased volume and decreased motility
A. Epididymis
B. Ductus deferens
C. Seminal vesicles
D. Prostate gland
E. Bulbourethral glands
intervals between collection of sperm sample following a 3-sample test
2-week interval
storage temp of sperm sample
37C or RT
Semen Analysis Parameters
AP2CMV
Appearance
pH
Sperm Concentration
Sperm Count
Sperm Motility
Sperm Morphology
Volume
Viscosity
this type of test may be useful to screen the sperm for WBC
leukocyte esterase reagent strip test
Red coloration of sperm may be due to
RBC
yellow coloration of sperm may be cause by
urine contam
prolonged abstinence
medications
urine affects the sperm motility
true or false
true
a fresh semen specimen is clotted and should liquefy within
30-60 mins after collection
what may be added to the semen if it does not liquefy after 2 hours?
alpha-chymotrypsin
Decreased volume after proper collection may be indicative of
infertility
consistency of the fluid and may be related to specimen liquefaction
viscosity
normal semen is
acidic or alkaline?
alkaline
Equation for Total Sperm Count
Sperm Concentration x Specimen Volume
Process of immobilizing the sperm for specimen count
Dilution
Diluting fluid for Sperm analysis
Sodium Bicarbonate and formalin (Traditional)
Saline and distilled water
stain that may be used to aid sperm count in using bright field microscopy
crystal violet
immature sperm and wbc are termed as
round cells
amount of leukocytes counted that is associated with infertility
>1million/mL
presence of >1 million spermatids/mL indicates
disruption of spermatogenesis
manner of reporting sperm concentration
million/mL
recommended the Neubauer chamber count for Sperm count
WHO
amount of semen used for assessing motility
10ul under 22x22mm cover slip
no. of fields evaluated for sperm motility
20 High-power fields
the presence of a high percentage of immobile sperm and clumps of sperm automatically indicate infertility
True or False
False, it requires further evaluation to determine sperm viability or the presence of sperm agglutinins
4 parts of the sperm that is evaluated in morphology
head
neckpiece
midpiece
tail
describe a normal sperm head
oval shaped
approx 5um long
3um wide
describe the length of sperm tail
45 um long
poor ovum penetration may be due to
abnormal head
abnormalities of these affect the motility of sperm
neckpiece
midpiece
tail
part of sperm that is critical to ovum penetration located at the tip of the head
acrosomal cap
approx size of the acrosomal cap
approx half of the head
2/3 of the sperm nucleus
Thickest part of the sperm tail
midpiece
surrounds the midpiece that produces the energy required by the tail for motility
mitochondrial sheath
best stain for Sperm
papanicolaou stain
how long are air dried slides stable?
24 hours
number of sperm that should be identified
200
method of sperm analysis wherein addt'l parameters are done
Kruger's strict criteria
Equation for counting round cells in Sperm analysis
it is suspected when a specimen has a normal sperm concentration with markedly decreased motility
Decreased Sperm Viability
what stain is mixed with semen to evaluate viability?
eosin-nigrosin
in Sperm viability test, which of the sperm (alive or dead) is infiltrated by the dye?
dead
Normal viability of sperm requires ?% living cells
75%
test used to detect presence of fructose in sperm
Resorcinol Test
normal quantitative level of fructose
=>13umol per ejaculate
how is quantitative measurement of sperm fructose done?
spectrophotometric method
what can be done to preserve the fructose content of sperm?
freezing
presence of these antibodies is suspected when clumps of sperm are observed during routine semen analysis
Antisperm Abs
2 tests to detect presence of antibody-coated sperm
mixed agglutination reaction
immunobead test
a screening procedure used primarily to detect the presence of IgG abs in semen
Mixed agglutination reaction
a more specific procedure in that it can be used to detect the presence of IgG, IgM, and IgA Abs and demonstrates what area of the sperm the autoAbs are affecting
Immunobead test
type of beads used in immunobead test
polyacrylamide beads
3 microbes most commonly suspected affecting sperm composition
Chlamydia trachomatis
Mycoplasma hominis
Ureaplasma urealyticum
chemicals present in semen that is usually tested
neutral alpha-glucosidase
zinc
citric acid
prostatic acid phosphatase
decreased neutral alpha-glucosidase suggests a disorder of the
epididymis
Decreased zinc, citrate, and Acid phosphatase indicated a lack of this fluid
prostatic fluid
determination of this may be used in cases of alleged rape
acid phosphatase
detection of seminal glycoprotein p30 (more specific)
the only concern in postvasectomy semen analysis
presence of spermatozoa
test involved in sperm function tests
hamster egg penetration assay
cervical mucus penetration test
hypo-osmotic swelling test
in vitro acrosome reaction
Maturation of spermatozoa takes place in the:
B.
Enzymes for the coagulation and liquefaction of semen are produced by the:
A.
The major component of seminal fluid is:
A.
If the first portion of a semen specimen is not collected,the semen analysis will have an abnormal:
B.
Failure of laboratory personnel to document the time a semen sample is collected primarily affects the interpretationof semen:
D.
Liquefaction of a semen specimen should take place within:
A.
A semen specimen delivered to the laboratory in a condom has a normal sperm count and markedly decreased sperm motility. This is indicative of:
D.
An increased semen ph may be caused by:
D.
Proteolytic enzymes may be added to semen specimens to:
A.
The normal sperm concentration is:
B.
Given the following information, calculate the sperm concentration:
dilution, 1:20
sperm counted in five RBC squares on each side of the hemocytometer, 80and 86 volume, 3 mL
D.
dilution, 1:20
volume, 3 mL
Using the above information, calculate the sperm concentration when 80 sperm are counted in 1 WBC square and 86 sperm are counted in another WBC square
C.
The primary reason to dilute a semen specimen before performing a sperm concentration is to:
A.
When performing a sperm concentration, 60 sperm are counted in the RBC squares on one side of the hemocytometer and 90 sperm are counted in the RBC squares on the other side. The specimen is diluted 1:20. The:
D.
Sperm motility evaluations are performed:
A.
Sperm motility is evaluated on the basis of:
D.
The percentage of sperm showing average motility that is considered normal is:
C.
All of the following are grading criteria for sperm motility except:
A.
The purpose of the acrosomal cap is:
B.
The sperm part containing a mitochondrial sheath is the:
D.
All of the following are associated with sperm motility except the:
B.
The morphologic shape of a normal sperm head is:
B.
Normal sperm morphology when using the WHO criteria is:
C.
Additional parameters measured by Kruger strict morphology include all of the following except:
B.
Round cells that are of concern and may be included in sperm counts and morphology analysis are:
D.
If 5 round cells per 100 sperm are counted in a sperm morphology smear and the sperm concentation is 30 million, the concentraton of round cells is:
B.
Following an abnormal sperm motility test with a normal sperm count, what additional test might be ordered?
C.
Follow-up testing for a low sperm concentration would include testing for:
C.
The immunobead test for antisperm antibodies:
D.
Measurement of alpha-glucosidase is performed to detect a disorder of the:
A.
A specimen delivered to the laboratory with a request for prostatic acid phosphatase and glycoprotein p30 was collected to determine:
A.
Following a negative postvasectomy wet preparation, the specimen should be:
A.
Standardization of procedures and reference values for semen analysis is primarily provided by the:
