SEMEN ANALYSIS Flashcards
1
Q
INDICATIONS
A
to check for infertility
to evaluate success of post vasectomy procedure
for Medico-legal cases/forensic studies
2
Q
site of spermatogenesis; produces 5% of semen volume (sperm cells)
A
Testis (seminiferous tubules)
3
Q
site of sperm maturation and storage
A
Epididymis
4
Q
provide nutrients(fructose) for sperm
A
Seminal Vesicles
5
Q
coagulation and liquefaction
A
Prostate gland
6
Q
produce acidic fluid that contains ACP, citric acid, zinc, proteolytic enzymes
A
Prostate gland
7
Q
add alkaline mucus to neutralize prostatic acid and vaginal acidity
A
Bulbourethral glands/ Cowper’s land
8
Q
produce testosterone
A
Leydig cells
9
Q
provide nutrients for the germ cells as they undergo spermatogenesis
A
Sertoli cells
10
Q
propel sperm to the ejaculatory duct
A
Vas deferens / ductus deferens
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%
12
Q
- produced by the seminiferous tubules of the testes
A
Sperm cells
13
Q
CHEMICAL CONSTITUENTS
A
Acid phosphatase
Zinc
Fructose
Potassium, citric acid, ascorbic acid
Proteolytic enzymes
Spermine and choline
14
Q
distinguishes semen from other fluids
A
Acid phosphatase
15
Q
major nutrient of spermatozoa
A
Fructose
16
Q
liquefaction and coagulation of seminal fluid
A
Proteolytic enzymes
17
Q
Abstinence period:
A
3 to 5 days or not more than 7 days
18
Q
Abstinence period Prolonged:
A
higher volume, decreased sperm motility and increased flavin content
19
Q
Time of collection:
A
preferably early in the morning with an empty bladder before ejaculation
20
Q
Container:
A
wide-mouthed, warm, sterile glass or plastic
21
Q
Transport and testing: brought and tested in the lab within
A
30 minutes to 1 hour
22
Q
Specimens for fructose level analysis should be tested within
A
2 hours
23
Q
Fertility testing: requires [?] at 2 weeks interval
A
2 to 3 samples
24
Q
is essential
A
Complete collection of semen
25
First portion not collected (1st and 2nd fractions)
Falsely decreased sperm count
Falsely increased pH
Specimen will not liquefy
26
Last portion not collected (3rd fraction)
Falsely decreased semen volume
Falsely increased sperm count
Falsely decreased pH
Specimen will not clot
27
METHODS OF COLLECTION
Self-Production or Masturbation
Condom Collection
Vaginal Vault Aspiration
Coitus Interruptus
28
: best method collection because it prevents contamination
Self-Production or Masturbation
29
: requires the use of condoms that are non-lubricant-containing and made from polyurethane.
Condom Collection
30
: contains spermicidal agents
***Ordinary condoms
31
: aspiration of seminal fluid from the vaginal vault after coitus
Vaginal Vault Aspiration
32
: not reliable method of collection because the first portion of the ejaculate may be lost
Coitus Interruptus
33
All semen specimen are potential reservoirs for
HIV and HEPATITIS virus
34
STANDARD PRECAUTIONS
35
Discarded as
biohazardous waste
36
Specimens must be kept at
body temperature (37oC)
37
: preserved in frozen state at -85C and stored for one year
Artificial insemination
38
: should be frozen if delay in testing is unavoidable
Fructose level testing
39
PHYSICAL EXAMINATION
Performed after liquefaction (within [?])
30 to 60 minutes
40
Color
Grayish white to Pearly white
41
Transparency
Translucent
42
Volume
2-5mL/ejaculation
43
Volume
Measured using a
graduated cylinder with 0.1 increments
44
Volume
Increased:
Decreased:
Increased: prolonged abstinence
Decreased: infertility
45
Odor
Fishy, distinct, “Chlorox-like”, Musty, acrid
46
Viscosity
Highly viscous, pours in droplets
47
Viscosity
Normal:
viscid to highly viscid
48
Viscosity
Reporting:
0 (watery) – 4 (gel-like)
49
Prolonged [?] indicates a possible deficiency in prostatic enzymes
Liquefaction
50
Assessed by pouring semen into a graduated glass (normal: drop by drop)
Liquefaction
51
Incomplete [?] impedes sperm motility and should be reported
Liquefaction
52
Liquefaction Time
30 minutes-1 hour
53
pH
7.2-8.0
54
: possible infection within the reproductive tract
↑pH (alkaline)
55
: increased prostatic fluid , ejaculatory duct obstruction, poorly developed seminal vesicles
↓pH (acidic)
56
Specific Gravity
1.027 to 1.032
57
may indicate bleeding
Rust to red-brown
58
may indicate urine contamination, antibiotics, prolonged abstinence or pyospermia
Yellowish
59
may indicated infection and presence of WBCs
Increased white turbidity
60
infertility
Clear
61
MICROSCOPIC
Sperm motility
Sperm count
Sperm concentration
Sperm morphology
Sperm viability
62
Main factor which affects the penetrating ability of the sperm cells through the cervical mucus barrier
SPERM MOTILITY
63
Performed on a well-mixed liquefied specimen within 1 hour from time of collection, under 20 HPF
SPERM MOTILITY
64
SPERM MOTILITY
A [?] should be used
positive displacement pipette
65
SPERM MOTILITY
Coverslip:
22x22 mm
66
Evaluate the movement and direction
SPERM MOTILITY
67
SPERM MOTILITY
Alternative procedure: Examine [?] and count the percentages of the different motile categories using a manual cell counter
200 sperm per slide
68
SPERM MOTILITY
Normal: A minimum motility of [?] with a rating of [?] within [?]
50%
2.0
1 hour
69
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.
50%
25%
70
Number of cells per ejaculate
SPERM COUNT
71
SPERM COUNT
Normal Value:
> 40 million/ejaculate
72
Number of cells per ml
SPERM CONCENTRATION
73
SPERM CONCENTRATION
Normal Value:
> 20 million/ml
74
NORMAL VALUES
Routine:
Kruger’s Strict Criteria:
>30% normal
>14% normal
75
involves measuring the head, neck, tail and acrosome
Kruger’s Strict Criteria
76
Diluting Fluids:
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
Only [?] should be counted
fully developed sperm cells
78
[?] must not be included
Immature sperm and WBCs (“round cells”)
79
[?] can be counted if clinically significant
Spermatids (immature sperm cells) and WBCs
80
– inflammation or infection of the reproductive system that can lead to infertility
>1 million WBCs per ml
81
– disruption of spermatogenesis
> 1 million spermatids per ml
82
Procedure: 1 drop of liquefied semen → pre-warmed slide → cover with a coverslip ringed with petrolatum
Hanging Drop Method
83
Count at least 200 spermatozoa under HPF
Hanging Drop Method
84
Motile: moving actively forward
Non-motile: sluggish in direction
Hanging Drop Method
85
Performed in an undiluted semen under approx 20 HPF
Routine Grading of Motility
86
Utilizes undiluted specimen
Makler
87
Sperm cells are immobilized by heating
Makler
88
Sperm cells are counted using the 4 corner squares and the large central square
Neubauer Hemocytometer
89
Both sides loaded
Neubauer Hemocytometer
90
Counts should agree within 10%
Neubauer Hemocytometer
91
Phase-contrast or bright-field microscopy
Neubauer Hemocytometer
92
- Decrease in the number of sperm cells or presence of few motile cells
Oligospermia
93
- seen in unilateral or bilateral hypotrophic testes and hypothyroidism
Oligospermia
94
- Complete or total absence of spermatozoa
Azospermia
95
- Seen in bilateral underdeveloped testes, obstruction and infection with gonorrhea
Azospermia
96
- Presence of dead or immobile sperm cells
Necrospermia
97
- Seen in hypertrophic testes and decrease in fructose levels
Necrospermia
98
- Oval shaped; measuring 5um in length and 3um in width
Head
99
- Acrosomal cap: should occupy 1/2 of the head and 2/3 of the nucleus
Head
100
- Attaches the tail to the head
Neck/Middle Piece
101
- Contains mitochondria that provides energy for flagellar motion
Neck/Middle Piece
102
- Approximately 45um in length and exhibits a whip-like motion that propels the sperm cell
Tail
103
Stain and evaluate at least
200 sperm cells under oil immersion field
104
Percentage of [?] is reported
abnormal sperm
105
Stains:
• Papanicolaou – stain of choice
• Giemsa stain
• Wright’s stain
106
Head:
double heads, giant and amorphous heads, pinheads, tapered heads and constricted heads
107
Tail:
doubled, coiled or bent
108
Neckpiece:
abnormally long
109
Provides objective testing sperm motility based on sperm velocity and trajectory (direction of motion)
Computer-Assisted Semen Analysis
110
Also determines sperm concentration and morphology
Computer-Assisted Semen Analysis
111
Differentiation and enumeration of round cells—
immature sperm cells(spermatids) and leukocytes
112
Variables:
• C = number of round cells
• N = number of spermatids and/or leukocytes counted
• S = sperm concentration in millions per ml
C = (N x S) / 100
113
Suspected if a specimen has normal sperm concentration with markedly decreased motility
I. Sperm Viability Test/Bloom’s test/ Eosin Negrosin
114
Assessed within 1 hour of ejaculation by mixing the specimen with Eosin-Nigrosin
I. Sperm Viability Test/Bloom’s test/ Eosin Negrosin
115
number of dead cells in 100 sperm is counted using a bright-field or phase contrast microscope
I. Sperm Viability Test/Bloom’s test/ Eosin Negrosin
116
I. Sperm Viability Test/Bloom’s test/ Eosin Negrosin
LIVING CELLS:
DEAD CELLS:
Normal:
bluish white in color
red against a purple background
75% living cells/100 sperm cells
117
Low levels are associated with androgen deficiency, low testosterone level and seminal vesicle insufficiency
Fructose Test
118
Samples tested within 2 hours or frozen to prevent fructolysis
Fructose Test
119
Resorcinol Test: 1 ml semen + 9 ml reagent --> boil --> orange red color
Fructose Test
120
Spectrophotometric: used to quantitate fructose level in semen
Fructose Test
121
Normal: ≥ 13 umol/ejaculate
Fructose Test
122
Destroys blood-testes barrier
Antisperm Antibodies (ASA)
123
Can be detected in men and women
Antisperm Antibodies (ASA)
124
screening procedure to detect IgG antibodies
Mixed agglutination reaction (MAR)
125
The semen sample is incubated with AHG and a suspension of latex particles or treated RBCs coated with IgG
Mixed agglutination reaction (MAR)
126
(+): >50% agglutination
Mixed agglutination reaction (MAR)
127
more specific test and detects IgG, IgM and IgA antibodies and demonstrate
Immunobead test
128
what area of the sperm the autoantibodies are affecting
(+): ≥ 20% sperm with bound immunobead
Immunobead test
129
Post-coital test to determine the quality of the cervical mucus and the ability of sperm cells to penetrate it
Sims-Huhner Test
130
Determines the presence of choline
Florence Test
131
Florence Test Reagents:
potassium iodide and iodine crystals
132
Florence Test (+) Result:
presence of brown rhombic or needle-like crystals under the microscope
133
Determines the presence of spermine
Barbiero’s Test
134
Barbiero’s Test Reagents:
picric acid and trichloroacetic acid
135
Barbiero’s Test (+) Result:
presence of yellow leaf-like structures
136
Determines the tenacity of the mucus
Spinbarkeit Test
137
Specimens are tested at monthly intervals, beginning at 2 months after vasectomy and continuing until two consecutive monthly specimens shows no spermatozoa
Post-vasectomy Semen Analysis
