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Semenalysis Flashcards

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

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2
Q

Spermatozoa:

§ Produced in the__________

A

seminiferous tubules of the testes

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3
Q

Spermatozoa:

§ Mature and stored in ____________

A

epididymis

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4
Q

§ Spermatozoa and fluid from epididymis contribute _________of semen volume

A

5%

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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:

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6
Q

Spermatozoa become mobile when exposed to ____________

A

seminal fluid

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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:

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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

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9
Q

Semen - Physiology

_____________:

§ Contribute 5% of semen volume

§ Thick, alkaline mucus (neutralize acidity from prostate secretions and vaginal acidity)

A

Bulbourethral glands

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10
Q

Specimen Collection

Specimens collected following a period of sexual abstinence ________

A

(3-5 days)

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11
Q

Specimen Collection

Fertility testing:____________; 2 abnormal samples considered significant

A

2-3 samples tested at 2- week intervals

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12
Q

Warm sterile glass or plastic container

Specimen kept at________ and delivered to lab within 1 hour of collection

A

RT

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13
Q

Specimen Collection

Record time of collection and specimen receipt

Fresh semen specimen is _________

A

clotted

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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

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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

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16
Q

Analysis:

Appearance ___________

A

Gray-white color, translucent, musty odor

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17
Q

Increased white turbidity:

______________________

A

presence of WBCs and infection within reproductive tract

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18
Q

Varying degrees of red coloration: __________

A

presence of RBCs

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19
Q

Analysis: Appearance

Yellow coloration: ______________

A

urine contamination, prolonged abstinence, medications

Note : Urine is toxic to sperm, thereby affecting evaluation of motility

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20
Q

Analysis: Volume

A

2-5 ml

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21
Q

Analysis: Volume

______________ following periods of extended abstinence

A

Increased volume:

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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:

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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

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24
Q

Analysis: Viscosity Ratings:

Note : Increased viscosity and incomplete liquefaction will impede sperm motility

A

0 (watery) to 4 (gel-like)

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25
Q

Analysis: pH ______________

A

Alkaline: 7.2 to 8.0

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26
Q

Increased pH: ______________________________

A

infection within reproductive tract

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27
Q

associated with increased prostatic fluid

A

Decreased pH:

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28
Q

What is used to test the sperm ph?

A

pH pad of urinalysis reagent strip

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29
Q

Analysis: Sperm Concentration/ Count

Normal sperm concentration: __________

A

> 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)

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30
Q

Analysis: Sperm Concentration/ Count

What is used to check the count of semen manually?

A

Neubauer counting chamber (amount of dilution and the # of squares counted vary among labs)

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31
Q

What is the dilution used in counting the sperm in Neubauer?

A

Commonly used dilution is 1:20; dilution is essential because it immobilizes the sperm prior to counting

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32
Q

Traditional diluting fluid contains ____________ and __________ or _____________ can also be used

A

Na bicarbonate

and formalin Saline

and distilled water

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33
Q

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

A

4 corner and center squares of the large center square (RBC counting area)

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34
Q

Analysis: Sperm Concentration/ Count

___________l is associated with inflammation or infection of the reproductive organs (can lead to infertility)

A

>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

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35
Q

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

A

1. dilution

  1. size and number of squares counted
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36
Q

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

A

Makler counting chamber

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37
Q

Analysis: Sperm Concentration/ Count

The method recommended by the WHO is the ______________

A

Neubauer chamber count

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38
Q

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

A

Sperm Motility

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39
Q

Analysis: Sperm Motility

% of sperm showing actual forward movement estimated after evaluating 20 HPFs

Motility is evaluated by both_ ___and _________.

A

speed and direction

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40
Q

4.0

**Grade a **

A

Rapid, straight-line motility

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41
Q

3.0

Grade b

A

Slower speed, some lateral movement

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42
Q

2.0

Grade b

A

Slow forward progression, noticeable lateral movement

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43
Q

1.0

Grade c

A

No forward progression

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44
Q

0

Grade d

A

No movement

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45
Q

Analysis: Sperm Motility

Normal: ____________________

A

minimum motility of 50% with a rating of 2.0 after 1 hour

46
Q

Analysis: Sperm Motility

WHO:

A

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
Q

Analysis: Sperm Motility

Presence of a________________ of sperm requires further evaluation to determine sperm viability or presence of sperm agglutinins

A

high % of immotile sperm and clumps

48
Q

Analysis: Sperm Motility

______________

Provides objective determination 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

A

Computer-assisted semen analysis (CASA)

49
Q

Analysis: Sperm Morpholog

y Morphology is evaluated with respect to the ____________, ______________ , _______ and __________

A

structure of head, neckpiece, midpiece and tail

50
Q

Normal sperm:

A

§ Oval-shaped head 5 um long, 3 um wide

§ Long, flagellar tail 45 um long

51
Q

§___________ attaches the head to the tail and midpiece

A

 Neckpiece

52
Q

§______________ is the thickest part of the tail

– surrounded by a mitochondrial sheath that produces the energy required by the tail for motility

53
Q

______________– critical to ovum penetration at the tip of head(enzyme-containing)

A

​§ Acrosomal cap

54
Q

Analysis: Sperm Morphology

Head abnormalities: _________________

A

poor ovum penetration

55
Q

Analysis: Sperm Morphology

Neckpiece, midpiece and tail abnormalities: ____________

A

affect motility

56
Q

_________________________ 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

A

Morphology

57
Q

Analysis: Sperm Morphology

Abnormalities in head structure: _________

A

double heads, giant and amorphous heads, pinheads, tapered heads, constricted heads

58
Q

Analysis: Sperm Morphology

Abnormal sperm tails: ____________

A

double, coiled, bent

59
Q

Abnormally long neckpiece:

A

may cause sperm head to bend backward and interfere with motility

60
Q

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

A

Kruger’s strict criteria

61
Q

What is the normal value for routine criteria? What about the strict criteria?

A

Analysis: Sperm Morphology

Normal Values: q >30% normal forms (routine criteria)

q >14% normal forms (strict criteria)

62
Q

Kruger Strict Guidelines

>=15% normal:

A

Normal range - Good prognosis

63
Q

Kruger Strict Guidelines

Sub optimal range - Prognosis is fair
to good, however, the lower the
percent normal, the lower the
chance of successful fertilization

A

5-14% normal:

64
Q

0-4% normal:

A

Poor prognosis - Will usually need
IVF with intracytoplasmic sperm
injection (ICSI)

65
Q

Additional Testing

A

Sperm viability
Seminal fluid fructose level
Sperm agglutinins
Microbial infection

66
Q

Sperm Viability

Decreased sperm viability is suspected when a specimen has a ________________

A

normal sperm concentration with decreased motility

67
Q

How is viability evaluated?

A

Viability is evaluated by mixing specimen with an eosin nigrosin stain, preparing a smear and counting # of dead cells in 100 sperm

68
Q

Sperm Viability

_______________ – not infiltrated by dye and remain a bluish-white color

A

Living cells

69
Q

Sperm Viability

Dead cells – stain ___________________

A

red against purple background

70
Q

Normal viability requires ________% living cells

71
Q

Seminal Fluid Fructose

Low sperm concentration maybe caused by _________________ - indicated by low to absent fructose level in semen

A

lack of support medium produced in seminal vesicles

72
Q

______________– to check for presence of fructose

A

Resorcinol test

73
Q

What is the color when fructose is present?

A

(orange-red color when fructose is present)

74
Q

FRUCTOSE TEST

Normal quantitative level: >= ________________

A

13 umol/ ejaculate

75
Q

Specimen tested within ________________ to prevent fructolysis

A

2 hours or frozen

76
Q

Present in both men and women

A

Antisperm Antibodies

77
Q

Where can you detect the antisperm antibodies?

A

Detected in

  1. semen,
  2. cervical mucosa,
  3. or serum
78
Q

Antisperm Ab Possible cause of infertility

___________are more frequently encountered

A

Male antisperm antibodies

79
Q

What is the reason behind antisperm antibodies?

A

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
Q

How is the antisperm demonstrated in the male?

A

Antisperm Antibodies

Presence of antibodies in the male: clumping of sperm

81
Q

How is the antisperm demonstrated in the female?

A

Presence of antisperm antibodies in the female (demonstrated by mixing semen with female cervical mucosa or serum and observing for agglutination):

82
Q

How can presence of antisperm antibodies be suspected?

A

normal semen analysis accompanied by continued infertility

83
Q

Tests to detect the presence of antibodycoated sperm:

A
  1. Mixed agglutination reaction (MAR) test
  2. Immunobead test
84
Q

Antisperm Antibodies Tests to detect the presence of antibodycoated sperm:

_________________ – screening procedure used primarily to detect the presence of IgG antibodies

A

Mixed agglutination reaction (MAR) test

85
Q

MAR test procedure

A

Semen sample containing motile sperm is incubated with IgG AHG and a suspension of latex particles or 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
Q

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

A

Immunobead test

87
Q

Immunobead test procedure

A

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
Q

What is the normal value for immunobead testing?

A

Presence of beads on < 20% of sperm is normal

89
Q

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 _________

A
  1. epididymis
  2. prostatic fluid
90
Q

Normal Semen Chemical Values

Neutral aglucosidase >=_________________

A

20 mU / ejaculate

91
Q

Normal Semen Chemical Values

Zinc >_________________

A

=2.4 umol/ ejaculate

92
Q

Normal Semen Chemical Values

Citric acid__________

A

>=52 umol/ ejaculate

93
Q

Normal Semen Chemical Values

Acid phosphatase____________________

A

>=200 units/ ejaculate

94
Q

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

A

For rape cases

95
Q

Postvasectomy Semen Analysis

Presence or absence of spermatozoa

Specimens are routinely tested at monthly intervals for?

A

Specimens are routinely tested at monthly intervals, beginning at 2 months postvasectomy and continuing until 2 consecutive monthly specimens show no sperm

96
Q

Sperm Function Tests

A
  1. Hamster egg penetration
  2. Cervical mucus
    penetration
  3. Hypo-osmotic
    swelling
  4. In vitro acrosome
    reaction
97
Q

Sperms are incubated with species-nonspecific hamster eggs and penetration is observed microscopically

A

Hamster egg penetration

98
Q

Observation of sperm penetration ability of partner’s midcycle cervical mucus

A

Cervical mucus penetration

99
Q

Sperm exposed to low sodium concentrations are evaluated for membrane integrity and sperm viability

A

Hypo-osmotic swelling

100
Q

Evaluation of the acrosome to produce enzymes essential for ovum penetration

A

In vitro acrosome reaction

101
Q

Normal Values

Volume ________________

102
Q

Normal Values

Viscosity

A

Pours in droplets

103
Q

Normal value:

pH

104
Q

Normal Values

Sperm concentration

A

>20 million/ml

105
Q

Normal Values

Sperm count

A

>40 million/ejaculate

106
Q

N. V

Motility

A

>50% within 1 hour

107
Q

Normal Value

Quality

108
Q

Normal Values

Morphology

A

Morphology

>14% normal forms (strict
criteria)
>30% normal forms
(routine criteria)

109
Q

Normal Values:

White blood cells _________

A

<1.0 million/ml

Laboratory Diagnosis (4 decks)

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