Semenalysis Flashcards
(110 cards)
1
Q
Semen - Physiology
Composed of 4 fractions contributed individually by___________, ______ ,___________ and _________
Mixing of all 4 fractions essential for production of normal semen specimen
A
testes and epididymis,
seminal vesicles,
prostate,
bulbourethral glands
2
Q
Spermatozoa:
§ Produced in the__________
A
seminiferous tubules of the testes
3
Q
Spermatozoa:
§ Mature and stored in ____________
A
epididymis
4
Q
§ Spermatozoa and fluid from epididymis contribute _________of semen volume
A
5%
5
Q
§ Contribute 60% of semen volume
§ Contain high content of fructose that the spermatozoa readily metabolize
§ Spermatozoa become mobile when exposed to seminal fluid
A
Seminal Vesicles:
6
Q
Spermatozoa become mobile when exposed to ____________
A
seminal fluid
7
Q
§ Contributes 20-30% of semen volume
§ Contains high concentrations of acid phosphatase, citric acid, zinc and proteolytic enzymes (responsible for coagulation and liquefaction of semen following ejaculation)
A
Prostate:
8
Q
§ The prostate contains high concentrations of _________, __________ ,_______ and __________-(responsible for coagulation and liquefaction of semen following ejaculation)
A
acid phosphatase, citric acid, zinc and proteolytic enzymes
9
Q
Semen - Physiology
_____________:
§ Contribute 5% of semen volume
§ Thick, alkaline mucus (neutralize acidity from prostate secretions and vaginal acidity)
A
Bulbourethral glands
10
Q
Specimen Collection
Specimens collected following a period of sexual abstinence ________
A
(3-5 days)
11
Q
Specimen Collection
Fertility testing:____________; 2 abnormal samples considered significant
A
2-3 samples tested at 2- week intervals
12
Q
Warm sterile glass or plastic container
Specimen kept at________ and delivered to lab within 1 hour of collection
A
RT
13
Q
Specimen Collection
Record time of collection and specimen receipt
Fresh semen specimen is _________
A
clotted
14
Q
Specimen Collection
Liquefy within ___________ after collection; failure to liquefy maybe due to a deficiency in prostatic enzymes
Analysis begins after liquefaction Specimen collected by masturbation or nonlubricant-containing polymeric silicone (Silastic) condoms
A
30-60 mins
15
Q
Semen Analysis Consists o
f macro and microscopic examination
Parameters reported include:
A
appearance,
volume,
viscosity,
pH,
sperm concentration
and count,
motility,
morphology
16
Q
Analysis:
Appearance ___________
A
Gray-white color, translucent, musty odor
17
Q
Increased white turbidity:
______________________
A
presence of WBCs and infection within reproductive tract
18
Q
Varying degrees of red coloration: __________
A
presence of RBCs
19
Q
Analysis: Appearance
Yellow coloration: ______________
A
urine contamination, prolonged abstinence, medications
Note : Urine is toxic to sperm, thereby affecting evaluation of motility
20
Q
Analysis: Volume
A
2-5 ml
21
Q
Analysis: Volume
______________ following periods of extended abstinence
A
Increased volume:
22
Q
l Associated with infertility - may indicate improper functioning of one of the semen-producing organs
l Incomplete specimen collection must also be considered
A
Decreased volume:
23
Q
Refers to consistency of fluid and may be related to specimen liquefaction
Normal specimen should be easily drawn into a pipette and form droplets that don’t appear clumped or stringy when discharged from the pipette
A
Analysis: Viscosity
24
Q
Analysis: Viscosity Ratings:
Note : Increased viscosity and incomplete liquefaction will impede sperm motility
A
0 (watery) to 4 (gel-like)
25
Analysis: pH \_\_\_\_\_\_\_\_\_\_\_\_\_\_
Alkaline: 7.2 to 8.0
26
Increased pH: \_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_
infection within reproductive tract
27
associated with increased prostatic fluid
Decreased pH:
28
What is used to test the sperm ph?
pH pad of urinalysis reagent strip
29
Analysis: Sperm Concentration/ Count
Normal sperm concentration: \_\_\_\_\_\_\_\_\_\_
\> 20 million sperm/ml
Note :
10-20 million/ml is borderline
Total sperm count/ejaculate = sperm concentration x specimen volume Normal: \> 40 million/ejaculate (20 M/ml x 2 ml)
30
Analysis: Sperm Concentration/ Count
What is used to check the count of semen manually?
Neubauer counting chamber (amount of dilution and the # of squares counted vary among labs)
31
What is the dilution used in counting the sperm in Neubauer?
Commonly used dilution is 1:20; dilution is essential because it **immobilizes the sperm prior to counting**
32
Traditional diluting fluid contains ____________ and __________ or _____________ can also be used
Na bicarbonate
and formalin Saline
and distilled water
33
Analysis: Sperm Concentration/ Count
Sperm cells are usually counted in the\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_
Both sides of hemocytometer are loaded and counted
Counts should agree within **10%**
Average of the 2 counts is used in the calculation
**Only fully developed sperm should be counted**
4 corner and center squares of the large center square (RBC counting area)
34
Analysis: Sperm Concentration/ Count
\_\_\_\_\_\_\_\_\_\_\_l is associated with inflammation or infection of the reproductive organs (can lead to infertility)
\>1 million WBCs/m
Note : The presence of \>1 million spermatids/ml indicates disruption of spermatogenesis – caused by viral infection, exposure to toxic chemicals, genetic disorders
35
Calculation of Sperm Concentration and Sperm Count
Calculation of sperm concentration is **dependent on\_\_\_\_\_\_\_\_\_\_**used and the **\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_.**
When using 1:20 dilution and counting 5 RBC squares in large center square: Sperm conc/ml = # of sperm x 1,000,000
Total sperm count = # of sperm/ml x specimen volume
**1. dilution**
2. size and number of squares counted
36
Analysis: Sperm Concentration/ Count
\_\_\_\_\_\_\_\_:
§ Provides a method for counting undiluted specimens
§ Sperm are **immobilized by heating part** of specimen prior to charging the chamber
§ **Sperm motility** evaluated using the **unheated portion**
Makler counting chamber
37
Analysis: Sperm Concentration/ Count
The method recommended by the WHO is the \_\_\_\_\_\_\_\_\_\_\_\_\_\_
Neubauer chamber count
38
Analysis: \_\_\_\_\_\_\_\_\_\_\_\_\_\_\_
Sperm capability of **forward, progressive movement** is critical for fertility
Undiluted specimen; well mixed, liquefied semen within 1 hour of specimen collection
Determine **% of motile sperm and quality of the motility**
Sperm Motility
39
Analysis: Sperm Motility
% of sperm showing a**ctual forward movement estimated after evaluating 20 HPFs**
Motility is evaluated by both\_ \_\_\_and \_\_\_\_\_\_\_\_\_.
speed and direction
40
**4.0**
**Grade a **
Rapid, straight-line motility
41
3.0
Grade b
Slower speed, some lateral movement
42
2.0
Grade b
Slow forward progression, noticeable lateral movement
43
1.0
Grade c
No forward progression
44
0
Grade d
No movement
45
Analysis: Sperm Motility
Normal: \_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_
minimum motility of 50% with a rating of 2.0 after 1 hour
46
Analysis: Sperm Motility
WHO:
Within 1 hour, 50% or more sperm should be motile in categories a, b, and c, or 25% or more should show progressive motility (a and b)
47
Analysis: Sperm Motility
Presence of a\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_ of sperm requires further evaluation to determine sperm viability or presence of sperm agglutinins
high % of immotile sperm and clumps
48
Analysis: Sperm Motility
\_\_\_\_\_\_\_\_\_\_\_\_\_\_
Provides **objective determinatio**n of **both sperm velocity** and **trajectory (direction of motion)**
Sperm concentration and morphology are also included in the analysis
Found primarily in laboratories that specialize in andrology and perform a high volume of semen analysis
Computer-assisted semen analysis (CASA)
49
Analysis: Sperm Morpholog
y Morphology is evaluated with respect to the \_\_\_\_\_\_\_\_\_\_\_\_, ______________ , _______ and \_\_\_\_\_\_\_\_\_\_
structure of head, neckpiece, midpiece and tail
50
Normal sperm:
§ Oval-shaped head 5 um long, 3 um wide
§ Long, flagellar tail 45 um long
51
§\_\_\_\_\_\_\_\_\_\_\_ attaches the head to the tail and midpiece
Neckpiece
52
§\_\_\_\_\_\_\_\_\_\_\_\_\_\_ is the thickest part of the tail
– surrounded by a mitochondrial sheath that **produces the energy required by the tail for motility**
Midpiece
53
\_\_\_\_\_\_\_\_\_\_\_\_\_\_– critical to ovum penetration at the tip of head(enzyme-containing)
§ Acrosomal cap
54
Analysis: Sperm Morphology
Head abnormalities: \_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_
poor ovum penetration
55
Analysis: Sperm Morphology
Neckpiece, midpiece and tail abnormalities: \_\_\_\_\_\_\_\_\_\_\_\_
affect motility
56
\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_ is evaluated from a thinly smeared, stained slide under oil immersion (Wright’s, Giemsa, Papanicolaou)
At least 200 sperm should be evaluated and % of abnormal sperm reported
Morphology
57
Analysis: Sperm Morphology
Abnormalities in head structure: \_\_\_\_\_\_\_\_\_
double heads, giant and amorphous heads, pinheads, tapered heads, constricted heads
58
Analysis: Sperm Morphology
Abnormal sperm tails: \_\_\_\_\_\_\_\_\_\_\_\_
double, coiled, bent
59
Abnormally long neckpiece:
may cause sperm head to bend backward and interfere with motility
60
Analysis: Sperm Morphology
q \_\_\_\_\_\_\_\_\_\_\_\_\_\_\_: requires use of **a stage micrometer or morphometry**
§ Measurement of head, neck and tail size
§ Size of acrosome
§ Presence of vacuoles
q Not routinely performed in the clinical laboratory **but is recommended by the WHO**
Kruger’s strict criteria
61
What is the normal value for routine criteria? What about the strict criteria?
Analysis: Sperm Morphology
Normal Values: q **\>30%** normal forms **(routine criteria)**
q **\>14%** normal forms **(strict criteria)**
62
Kruger Strict Guidelines
\>=15% normal:
Normal range - Good prognosis
63
Kruger Strict Guidelines
**Sub optimal range** - Prognosis is **fair**
to good, however, the lower the
percent normal, the lower the
chance of successful fertilization
5-14% normal:
64
0-4% normal:
**Poor prognosis** - Will usually need
IVF with intracytoplasmic sperm
injection (ICSI)
65
Additional Testing
Sperm viability
Seminal fluid fructose level
Sperm agglutinins
Microbial infection
66
Sperm Viability
Decreased sperm viability is suspected when a specimen has a \_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_
normal sperm concentration with decreased motility
67
How is viability evaluated?
Viability is evaluated by **mixing specimen** with an **eosin nigrosin stain**, preparing a smear and counting **# of dead cells in 100 sperm**
68
Sperm Viability
\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_ – not infiltrated by dye and **remain a bluish-white color**
Living cells
69
Sperm Viability
## Footnote
Dead cells – stain \_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_
red against purple background
70
Normal viability requires \_\_\_\_\_\_\_\_% living cells
75%
71
Seminal Fluid Fructose
Low sperm concentration maybe caused by _________________ - indicated by **low to absent fructose level** in **semen**
lack of support medium produced in seminal vesicles
72
\_\_\_\_\_\_\_\_\_\_\_\_\_\_– to check for presence of fructose
Resorcinol test
73
What is the color when fructose is present?
**(orange-red** color when fructose is present)
74
FRUCTOSE TEST
Normal quantitative level: \>= \_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_
13 umol/ ejaculate
75
Specimen tested within ________________ to prevent fructolysis
2 hours or frozen
76
Present in **both men and women**
Antisperm Antibodies
77
Where can you detect the antisperm antibodies?
Detected in
1. **semen,**
2. **cervical mucosa,**
3. **or serum**
78
Antisperm Ab Possible cause of infertility
\_\_\_\_\_\_\_\_\_\_\_are more frequently encountered
Male antisperm antibodies
79
What is the reason behind antisperm antibodies?
Antisperm Antibodies
**Under normal conditions**, **blood-testes barrier**
**separates sperm from the male immune
system**
When barrier is disrupted **(surgery,
vasectomy reversal, trauma, infection),**
antigens on sperm produce an immune
response that damages the sperm
Damaged sperm may cause production of
antibodies in the female partner
80
How is the antisperm demonstrated in the male?
Antisperm Antibodies
Presence of antibodies in the male: **clumping of sperm**
81
How is the antisperm demonstrated in the female?
Presence of antisperm antibodies in the female **(demonstrated by mixing semen with female cervical mucosa or serum and observing for agglutination)**:
82
How can presence of antisperm antibodies be suspected?
normal semen analysis accompanied by **continued infertility**
83
Tests to detect the presence of antibodycoated sperm:
1. Mixed agglutination reaction (MAR) test
2. Immunobead test
84
Antisperm Antibodies Tests to detect the presence of antibodycoated sperm:
\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_ – **screening procedur**e used primarily to detect the **presence of *IgG* antibodies**
Mixed agglutination reaction (MAR) test
85
MAR test procedure
Semen sample containing motile sperm is **incubated with IgG AHG** and a **suspension of latex particles** o**r treated RBCs coated** with **IgG Bivalent**
AHG binds simultaneously to both Ab on sperm and Ab on latex particles or RBCs, **forming microscopically visible clumps of sperm** and particles or **cells \<10 % of motile sperm attached to particles is normal**
86
Antisperm Antibodies Tests to detect the presence of antibodycoated sperm:
§ **More specific procedure**
§ Detects presence of **IgG, IgM, and IgA** antibodies
§ **Demonstrates what area** of the sperm the autoantibodies are affecting
Immunobead test
87
Immunobead test procedure
Sperm mixed with **polyacrylamide beads coated** with either **anti-IgG, anti-IgM, or anti- IgA**
Microscopic exam will **show beads attached to sperm at particular areas**
Reported as **“IgM tail antibodies**,” **“IgG head antibodies**,” etc. Presence of beads on \< 20% of sperm is normal
88
What is the normal value for immunobead testing?
Presence of beads on **\< 20%** of sperm is normal
89
Microbial and Chemical Testing
Routine aerobic and anerobic cultures
Additional chemical tests:
determination of levels of neutral alpha-glucosidase, Zn, citric acid, acid phosphatase
**Decreased alpha-glucosidase** suggests a disorder of \_\_\_\_\_\_\_\_\_\_\_\_\_-
**Decreased Zn, citrate, acid phosphatase** indicate lack of \_\_\_\_\_\_\_\_\_
1. epididymis
2. prostatic fluid
90
Normal Semen Chemical Values
Neutral aglucosidase \>=\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_
20 mU / ejaculate
91
Normal Semen Chemical Values
Zinc \>\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_
=2.4 umol/ ejaculate
92
Normal Semen Chemical Values
Citric acid\_\_\_\_\_\_\_\_\_\_
\>=52 umol/ ejaculate
93
Normal Semen Chemical Values
Acid phosphatase\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_
\>=200 units/ ejaculate
94
Microbial and Chemical Testing
Seminal fluid contains a **high concentration of prostatic acid phosphatase**, therefore the **detection of this enzyme can aid in** determining the **presence of semen in a specimen**
For rape cases
95
Postvasectomy Semen Analysis
Presence or absence of spermatozoa
Specimens are routinely tested at monthly intervals for?
Specimens are routinely tested at **monthly intervals**, beginning at **2 months postvasectomy** and **continuing until 2 consecutive monthly specimens show no sperm**
96
Sperm Function Tests
1. Hamster egg penetration
2. Cervical mucus
penetration
3. Hypo-osmotic
swelling
4. In vitro acrosome
reaction
97
Sperms are incubated with species-nonspecific hamster eggs and penetration is **observed microscopically**
Hamster egg penetration
98
Observation of sperm penetration ability of **partner’s midcycle cervical mucus**
Cervical mucus penetration
99
Sperm **exposed to low sodium concentrations** are evaluated for **membrane integrity and sperm viability**
Hypo-osmotic swelling
100
Evaluation of the acrosome to **produce enzymes essential for ovum penetration**
In vitro acrosome reaction
101
Normal Values
Volume \_\_\_\_\_\_\_\_\_\_\_\_\_\_\_\_
2-5 ml
102
Normal Values
Viscosity
Pours in droplets
103
Normal value:
pH
7.2-8.0
104
Normal Values
Sperm concentration
\>20 million/ml
105
Normal Values
Sperm count
\>40 million/ejaculate
106
N. V
Motility
\>50% within 1 hour
107
Normal Value
Quality
\>2.0
108
Normal Values
Morphology
Morphology
\>14% normal forms (**strict
criteria)**
\>30% normal forms
**(routine criteria)**
109
Normal Values:
White blood cells \_\_\_\_\_\_\_\_\_
\<1.0 million/ml
110
