SEMEN ANALYSIS Flashcards

1
Q

INDICATIONS

A

to check for infertility
to evaluate success of post vasectomy procedure
for Medico-legal cases/forensic studies

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2
Q

site of spermatogenesis; produces 5% of semen volume (sperm cells)

A

Testis (seminiferous tubules)

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3
Q

site of sperm maturation and storage

A

Epididymis

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4
Q

provide nutrients(fructose) for sperm

A

Seminal Vesicles

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5
Q

coagulation and liquefaction

A

Prostate gland

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6
Q

produce acidic fluid that contains ACP, citric acid, zinc, proteolytic enzymes

A

Prostate gland

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7
Q

add alkaline mucus to neutralize prostatic acid and vaginal acidity

A

Bulbourethral glands/ Cowper’s land

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8
Q

produce testosterone

A

Leydig cells

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9
Q

provide nutrients for the germ cells as they undergo spermatogenesis

A

Sertoli cells

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10
Q

propel sperm to the ejaculatory duct

A

Vas deferens / ductus deferens

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11
Q

COMPOSITION
Spermatozoa/Semen:
Seminal fluid/Seminal vesicles:
Prostate fluid.Prostate gland:
Alkaline mucus:
Epididymis, vas deferens, bulbourethral glands:

A

Spermatozoa/Semen: 5%
Seminal fluid/Seminal vesicles: 60-70%
Prostate fluid.Prostate gland: 20-30%
Alkaline mucus: 5%
Epididymis, vas deferens, bulbourethral glands: 10-15%

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12
Q
  • produced by the seminiferous tubules of the testes
A

Sperm cells

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13
Q

CHEMICAL CONSTITUENTS

A

Acid phosphatase
Zinc
Fructose
Potassium, citric acid, ascorbic acid
Proteolytic enzymes
Spermine and choline

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14
Q

distinguishes semen from other fluids

A

Acid phosphatase

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15
Q

major nutrient of spermatozoa

A

Fructose

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16
Q

liquefaction and coagulation of seminal fluid

A

Proteolytic enzymes

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17
Q

Abstinence period:

A

3 to 5 days or not more than 7 days

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18
Q

Abstinence period Prolonged:

A

higher volume, decreased sperm motility and increased flavin content

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19
Q

Time of collection:

A

preferably early in the morning with an empty bladder before ejaculation

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20
Q

Container:

A

wide-mouthed, warm, sterile glass or plastic

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21
Q

Transport and testing: brought and tested in the lab within

A

30 minutes to 1 hour

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22
Q

Specimens for fructose level analysis should be tested within

A

2 hours

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23
Q

Fertility testing: requires [?] at 2 weeks interval

A

2 to 3 samples

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24
Q

is essential

A

Complete collection of semen

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25
Q

First portion not collected (1st and 2nd fractions)

A

Falsely decreased sperm count
Falsely increased pH
Specimen will not liquefy

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26
Q

Last portion not collected (3rd fraction)

A

Falsely decreased semen volume
Falsely increased sperm count
Falsely decreased pH
Specimen will not clot

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27
Q

METHODS OF COLLECTION

A

Self-Production or Masturbation
Condom Collection
Vaginal Vault Aspiration
Coitus Interruptus

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28
Q

: best method collection because it prevents contamination

A

Self-Production or Masturbation

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29
Q

: requires the use of condoms that are non-lubricant-containing and made from polyurethane.

A

Condom Collection

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30
Q

: contains spermicidal agents

A

***Ordinary condoms

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31
Q

: aspiration of seminal fluid from the vaginal vault after coitus

A

Vaginal Vault Aspiration

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32
Q

: not reliable method of collection because the first portion of the ejaculate may be lost

A

Coitus Interruptus

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33
Q

All semen specimen are potential reservoirs for

A

HIV and HEPATITIS virus

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34
Q

STANDARD PRECAUTIONS

A
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35
Q

Discarded as

A

biohazardous waste

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36
Q

Specimens must be kept at

A

body temperature (37oC)

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37
Q

: preserved in frozen state at -85C and stored for one year

A

Artificial insemination

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38
Q

: should be frozen if delay in testing is unavoidable

A

Fructose level testing

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39
Q

PHYSICAL EXAMINATION

Performed after liquefaction (within [?])

A

30 to 60 minutes

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40
Q

Color

A

Grayish white to Pearly white

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41
Q

Transparency

A

Translucent

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42
Q

Volume

A

2-5mL/ejaculation

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43
Q

Volume
Measured using a

A

graduated cylinder with 0.1 increments

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44
Q

Volume
Increased:
Decreased:

A

Increased: prolonged abstinence
Decreased: infertility

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45
Q

Odor

A

Fishy, distinct, “Chlorox-like”, Musty, acrid

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46
Q

Viscosity

A

Highly viscous, pours in droplets

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47
Q

Viscosity
Normal:

A

viscid to highly viscid

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48
Q

Viscosity
Reporting:

A

0 (watery) – 4 (gel-like)

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49
Q

Prolonged [?] indicates a possible deficiency in prostatic enzymes

A

Liquefaction

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50
Q

Assessed by pouring semen into a graduated glass (normal: drop by drop)

A

Liquefaction

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51
Q

Incomplete [?] impedes sperm motility and should be reported

A

Liquefaction

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52
Q

Liquefaction Time

A

30 minutes-1 hour

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53
Q

pH

A

7.2-8.0

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54
Q

: possible infection within the reproductive tract

A

↑pH (alkaline)

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55
Q

: increased prostatic fluid , ejaculatory duct obstruction, poorly developed seminal vesicles

A

↓pH (acidic)

56
Q

Specific Gravity

A

1.027 to 1.032

57
Q

may indicate bleeding

A

Rust to red-brown

58
Q

may indicate urine contamination, antibiotics, prolonged abstinence or pyospermia

A

Yellowish

59
Q

may indicated infection and presence of WBCs

A

Increased white turbidity

60
Q

infertility

A

Clear

61
Q

MICROSCOPIC

A

Sperm motility
Sperm count
Sperm concentration
Sperm morphology
Sperm viability

62
Q

Main factor which affects the penetrating ability of the sperm cells through the cervical mucus barrier

A

SPERM MOTILITY

63
Q

Performed on a well-mixed liquefied specimen within 1 hour from time of collection, under 20 HPF

A

SPERM MOTILITY

64
Q

SPERM MOTILITY
A [?] should be used

A

positive displacement pipette

65
Q

SPERM MOTILITY
Coverslip:

A

22x22 mm

66
Q

Evaluate the movement and direction

A

SPERM MOTILITY

67
Q

SPERM MOTILITY
Alternative procedure: Examine [?] and count the percentages of the different motile categories using a manual cell counter

A

200 sperm per slide

68
Q

SPERM MOTILITY
Normal: A minimum motility of [?] with a rating of [?] within [?]

A

50%
2.0
1 hour

69
Q

WHO: within 1 hour, [?] or more of the sperm should be motile in categories a, b, and c, or [?] or more should show categories a and b.

A

50%
25%

70
Q

Number of cells per ejaculate

A

SPERM COUNT

71
Q

SPERM COUNT
Normal Value:

A

> 40 million/ejaculate

72
Q

Number of cells per ml

A

SPERM CONCENTRATION

73
Q

SPERM CONCENTRATION
Normal Value:

A

> 20 million/ml

74
Q

NORMAL VALUES
Routine:
Kruger’s Strict Criteria:

A

> 30% normal
14% normal

75
Q

involves measuring the head, neck, tail and acrosome

A

Kruger’s Strict Criteria

76
Q

Diluting Fluids:

A

1.5% sodium bicarbonate
2.1% formalin
0.5% chlorazene
4.1% formalin in sodium bicarbonate
5% sodium bicarbonate + 1% phenol in distilled H2O
Chilled distilled H2O

77
Q

Only [?] should be counted

A

fully developed sperm cells

78
Q

[?] must not be included

A

Immature sperm and WBCs (“round cells”)

79
Q

[?] can be counted if clinically significant

A

Spermatids (immature sperm cells) and WBCs

80
Q

– inflammation or infection of the reproductive system that can lead to infertility

A

> 1 million WBCs per ml

81
Q

– disruption of spermatogenesis

A

> 1 million spermatids per ml

82
Q

Procedure: 1 drop of liquefied semen → pre-warmed slide → cover with a coverslip ringed with petrolatum

A

Hanging Drop Method

83
Q

Count at least 200 spermatozoa under HPF

A

Hanging Drop Method

84
Q

Motile: moving actively forward
Non-motile: sluggish in direction

A

Hanging Drop Method

85
Q

Performed in an undiluted semen under approx 20 HPF

A

Routine Grading of Motility

86
Q

 Utilizes undiluted specimen

A

Makler

87
Q

 Sperm cells are immobilized by heating

A

Makler

88
Q

 Sperm cells are counted using the 4 corner squares and the large central square

A

Neubauer Hemocytometer

89
Q

 Both sides loaded

A

Neubauer Hemocytometer

90
Q

 Counts should agree within 10%

A

Neubauer Hemocytometer

91
Q

 Phase-contrast or bright-field microscopy

A

Neubauer Hemocytometer

92
Q
  • Decrease in the number of sperm cells or presence of few motile cells
A

Oligospermia

93
Q
  • seen in unilateral or bilateral hypotrophic testes and hypothyroidism
A

Oligospermia

94
Q
  • Complete or total absence of spermatozoa
A

Azospermia

95
Q
  • Seen in bilateral underdeveloped testes, obstruction and infection with gonorrhea
A

Azospermia

96
Q
  • Presence of dead or immobile sperm cells
A

Necrospermia

97
Q
  • Seen in hypertrophic testes and decrease in fructose levels
A

Necrospermia

98
Q
  • Oval shaped; measuring 5um in length and 3um in width
A

Head

99
Q
  • Acrosomal cap: should occupy 1/2 of the head and 2/3 of the nucleus
A

Head

100
Q
  • Attaches the tail to the head
A

Neck/Middle Piece

101
Q
  • Contains mitochondria that provides energy for flagellar motion
A

Neck/Middle Piece

102
Q
  • Approximately 45um in length and exhibits a whip-like motion that propels the sperm cell
A

Tail

103
Q

Stain and evaluate at least

A

200 sperm cells under oil immersion field

104
Q

Percentage of [?] is reported

A

abnormal sperm

105
Q

Stains:

A

• Papanicolaou – stain of choice
• Giemsa stain
• Wright’s stain

106
Q

Head:

A

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

107
Q

Tail:

A

doubled, coiled or bent

108
Q

Neckpiece:

A

abnormally long

109
Q

 Provides objective testing sperm motility based on sperm velocity and trajectory (direction of motion)

A

Computer-Assisted Semen Analysis

110
Q

 Also determines sperm concentration and morphology

A

Computer-Assisted Semen Analysis

111
Q

Differentiation and enumeration of round cells—

A

immature sperm cells(spermatids) and leukocytes

112
Q

Variables:
• C = number of round cells
• N = number of spermatids and/or leukocytes counted
• S = sperm concentration in millions per ml

A

C = (N x S) / 100

113
Q

Suspected if a specimen has normal sperm concentration with markedly decreased motility

A

I. Sperm Viability Test/Bloom’s test/ Eosin Negrosin

114
Q

Assessed within 1 hour of ejaculation by mixing the specimen with Eosin-Nigrosin

A

I. Sperm Viability Test/Bloom’s test/ Eosin Negrosin

115
Q

number of dead cells in 100 sperm is counted using a bright-field or phase contrast microscope

A

I. Sperm Viability Test/Bloom’s test/ Eosin Negrosin

116
Q

I. Sperm Viability Test/Bloom’s test/ Eosin Negrosin
LIVING CELLS:
DEAD CELLS:
Normal:

A

bluish white in color
red against a purple background
75% living cells/100 sperm cells

117
Q

Low levels are associated with androgen deficiency, low testosterone level and seminal vesicle insufficiency

A

Fructose Test

118
Q

Samples tested within 2 hours or frozen to prevent fructolysis

A

Fructose Test

119
Q

Resorcinol Test: 1 ml semen + 9 ml reagent –> boil –> orange red color

A

Fructose Test

120
Q

Spectrophotometric: used to quantitate fructose level in semen

A

Fructose Test

121
Q

Normal: ≥ 13 umol/ejaculate

A

Fructose Test

122
Q

Destroys blood-testes barrier

A

Antisperm Antibodies (ASA)

123
Q

Can be detected in men and women

A

Antisperm Antibodies (ASA)

124
Q

screening procedure to detect IgG antibodies

A

Mixed agglutination reaction (MAR)

125
Q

The semen sample is incubated with AHG and a suspension of latex particles or treated RBCs coated with IgG

A

Mixed agglutination reaction (MAR)

126
Q

(+): >50% agglutination

A

Mixed agglutination reaction (MAR)

127
Q

more specific test and detects IgG, IgM and IgA antibodies and demonstrate

A

Immunobead test

128
Q

what area of the sperm the autoantibodies are affecting
(+): ≥ 20% sperm with bound immunobead

A

Immunobead test

129
Q

Post-coital test to determine the quality of the cervical mucus and the ability of sperm cells to penetrate it

A

Sims-Huhner Test

130
Q

Determines the presence of choline

A

Florence Test

131
Q

Florence Test Reagents:

A

potassium iodide and iodine crystals

132
Q

Florence Test (+) Result:

A

presence of brown rhombic or needle-like crystals under the microscope

133
Q

Determines the presence of spermine

A

Barbiero’s Test

134
Q

Barbiero’s Test Reagents:

A

picric acid and trichloroacetic acid

135
Q

Barbiero’s Test (+) Result:

A

presence of yellow leaf-like structures

136
Q

Determines the tenacity of the mucus

A

Spinbarkeit Test

137
Q

Specimens are tested at monthly intervals, beginning at 2 months after vasectomy and continuing until two consecutive monthly specimens shows no spermatozoa

A

Post-vasectomy Semen Analysis