Seminal Fluid Flashcards
(140 cards)
1
Q
What are the application of analysis of semen?
A
a. Andrology
b. Assisted Reproductive Technology
c. Fertility Studies
d. Post-vasectomy
e. Forensic analysis
2
Q
Determine the need for in-vitro fertilization
A
Andrology
3
Q
Combination of four secretions from structures of male
reproductive system.
These secretions are important for a normal semen specimen
A
Spermatozoa 5%
Seminal fluid 60-70%
Prostatic fluid 20-30%
Secretions from bulbourethral gland 5%
4
Q
Paired glands in the scrotum where sperm is produced
A
Testes
5
Q
The production of sperm is called
A
spermatogenesis
6
Q
✓ External location of the scrotum contributes to a __________ e that is optimal for sperm development
A
a lower scrotum temperature
7
Q
secrete sperm
A
seminiferous tubules
8
Q
Spermatogenesis:
A
- Mitosis of germ cells located in the epithelial cells
- Germ cells undergo meiosis (spermatogenesis)
- Development of immature (nonmotile) spermatozoa called
spermatids - Spermatids mature in the epididymis for 90 days or until
ejaculated
9
Q
provide nutrients and
support to the germ cells
A
Sertoli cells or nurse cells
10
Q
Semen is ejaculated via
A
the vas deferens to the
ejaculatory duct
11
Q
Receive both the mature sperm and the secretions from the
seminal vesicle
A
ejaculatory duct
12
Q
Produce the most fluid in the semen (60-80%)
➢ Provides medium for sperm
A
seminal vesicle
13
Q
✓ Provides energy for the sperm to move
✓ Absence = immotile sperm
A
Fructose
14
Q
✓ Gray appearance of semen
✓ Blue to yellow fluorescence when semen is
viewed under woods lamp (UV light)
A
Flavin (pigment)
15
Q
Seminal vasicle contains?
A
Fructose
Flavin
Coagulation protiens
16
Q
Located below the bladder
Function: secretion of prostatic fluid & help propel the semen
during ejaculation by contracting
A
Prostate gland
17
Q
➢ 20-30% of the semen
➢ Milky & acidic
➢ Responsible for liquefaction of semen
A
Prostatic fluid
18
Q
Prostatic fluid consist of high concentrations of?
A
Acid phosphatase
citric acid
zinc
proteolytic enzymes
19
Q
Located below the prostate
Function: secretion of alkaline mucus
A
Bulbourethral gland
20
Q
➢ 5% of the semen
➢ Neutralize acidity of prostatic secretions and acidic
environment of vagina
A
Secretion of bulbourethral gland
21
Q
Acidic environment =
A
immobilize the sperm
22
Q
Involves both macroscopic & microscopic analysis of sperm
and semen
A
semenalysis
23
Q
7 parameters:
A
a. Appearance
b. Volume
c. Viscosity
d. pH
e. Sperm concentration & sperm count
f. Motility
g. Morphology
24
Q
Contains the highest number of
sperm & prostatic fluid
A
First part of semen
25
▪ Failure to collect first part of the semen =
Decrease in sperm concentration
Increased pH & Failure to liquefy
26
▪ Failure to collect last part of semen =
False decreased in volume
decreased in pH
specimen unable to clot
27
Prolonged abstinence have higher volume but decreased motility (HOW MANY DAYS)
2-7 days of abstinence
28
➢ WHO recommends 2-3 collections with at least 1-3
weeks apart
➢ 2 out 3 abnormal specimen = significant
Fertility testing
29
Place of collection:
hospital/laboratory premises
(recommended)
30
If not possible, semen specimen must be delivered:
within 1 hour & transported @ room temp
31
Specimen awaiting analysis:
37 degrees celsius
32
Manner of collection:
a. Masturbation (recommended)
b. Nonspermicidial, nonlubricant-containing rubber or
polyurethane condoms
33
contain spermicides
Ordinary condoms
34
= first portion of the
ejaculate may be lost and the low pH of the vaginal fluid may
affect sperm motility
Coitus interruptus
35
After collection, the lab personnel must take note of the
following:
1. Patient information (name, age, birthday)
2. Period of abstinence
3. Time of collection
4. Other abnormalities in the specimen quality
36
Normal appearance of semen
Gray-white color, appears translucent and is musty in
odor
37
✓ Presence of WBCs and infection
✓ WBC may be confused with
spermatids = leukocyte esterase to
screen for WBC
White turbidity
38
✓ Presence of RBC
Red coloration
39
✓ Urine contamination or
prolonged abstinence
✓ Urine = toxic to sperm; affects
motility
Yellow
40
Initially, semen appears as gel-like fluid that will liquefy within
30-60 minutes
41
Failure to liquefy =
prostatic enzyme deficiencies
42
Analysis of specimen cannot be done unless semen has
liquefied (T/F)
T
43
➢ If semen fails to liquefy within 2 hours, induce liquefaction
by:
1. Dulbecco’s phosphate-buffered saline (DPBS)
2. proteolytic enzymes, such as alpha-chymotrypsin or
bromelain
44
Normal VOLUME OF SEMEN
2-5 mL
45
Volume of semen is measured using?
graduated cylinder
46
prolonged abstinence =
Increased volume:
47
Decreased volume =
✓ Impaired function of semen-producing organs (seminal
vesicle)
✓ Infertility
✓ Incomplete collection
48
Consistency of the semen; associated with liquefaction
VISCOSITY
49
Normal viscosity of semen
Easily drawn into pipette & form droplets that do not
appear clumped or stringy
50
Abnormal viscosity of semen
Incompletely liquefied semen
✓ Clumped and are highly viscous
✓ Droplets that form threads longer than 2cm
51
Manner of reporting:
✓ 0 (watery) to 4 (gel-like)
✓ Low, normal or high
52
Normal pH of semen
7.2 – 8.0 (alkaline)
53
??? = Alkaline pH
Prostatic
Fluid
(Acidic)
+
Secretions from
bulbourethral
gland (Alkaline)
+
Secretions
from seminal
vesicle
(Alkaline)
54
Increased pH =
infection
55
Decreased pH =
increased prostatic fluid, obstruction or poorly
developed seminal vesicles
56
Testing used for semen
✓ Reagent pad for pH
✓ pH paper
✓ pH meter
57
Number of spermatozoa
per mL of semen
Sperm concentration
58
Spermatozoa per
ejaculate
Sperm count
59
Normal sperm conc
>20 to 250 million
per mL
60
Borderline low: sperm conc
10-20
million per mL
61
Counted using (sperm conc)
Neubauer
counting chamber
62
Computed by (sperm count)
sperm
concentration x volume
63
Normal sperm count
>40 million/ejaculate
64
Neubauer counting chamber steps
1. Dilute the semen with diluting fluid (1:20 standard dilution)
2. Load the diluted sample to both sides of Neubauer counting
chamber
3. Stand for 3-5 mins to settle
4. Use the chamber for RBC counting
5. Only count mature sperm
65
Traditional diluting fluid
sodium bicarbonate and
formalin
66
Alternative diluting fluid
Saline and cold water
67
If counts do not agree within 10% margin...
REPEAT dilution and
counting
68
what cells may be significant
Round cells
69
>1 million leukocytes/mL is significant and is associated with
infection (prostatitis) & poor sperm
quality
70
>1 million spermatids/mL indicates
disruption of spermatogenesis
71
are the predominant form
of leukocytes in the semen
Peroxidase-positive granulocytes
72
= differentiate peroxidasepositive
granulocytes from spermatids & lymphocytes
Peroxidase stain
73
computation of sperm concentration:
If standard dilution (1:20) is used, multiply the
average number counted to 1 million
74
computation of sperm count:
sperm conc x volume
75
Progressive, forward motility
sperm motility
76
Method of sperm motility
1. Place consistent volume (10uL) of semen on glass slide
2. Evaluate percentage of sperm w/ forward movement in at
least 20 HPF
3. Another way: examine 200 spermatozoa and count the
percentage of sperm with different speed and direction
using cell counters
77
Normal: >50% of sperm with a
2.0 grading
Motility w/ speed & direction
78
Does not include speed
WHO 2010 Alternative Grading
79
Motile with rapid, straight line motility
4.0; a
80
motilr with slower speed
3.0; b
81
motile with slow forward progression
2.0; b
82
motile without forward progression
1.0; c
83
no movement
0; d
84
sperm moving linearly or in a large circle
progressive motility (PM)
85
sperm moving with an absence of progressing
nonprogressive motility (NP)
86
no movement
Immotility (IM)
87
High percentage of immotile sperm =
further evaluation to
determine vitality or presence of sperm agglutinins
88
Abnormal sperm morphology can also cause
INFERTILITY
89
Sperm morphology is evaluated by the structure of
head,
midpiece and tail
90
Contains the acrosomal cap
✓ 2/3 of the head
✓ Important for ovum penetration
HEAD
91
Thickest part of sperm due to mitochondrial sheet
✓ Provides energy for the tail
✓ Abnormal midpiece may cause the head to bend
backward
MIDPIECE
92
Method FOR SPERM MORPHOLOGY
➢ Prepare a smear using 10uL of semen
➢ Stain with Wright’s, Giemsa, Shorr, or Papanicolaou stain
➢ At least 200 spermatozoa should be evaluated and the
percentage of normal and abnormal sperm is reported
93
Reporting FOR SPERM MORPHOLOGY
Normal: >30% in normal forms
94
✓ Stricter evaluation used in assisted reproduction
✓ Include additional parameters: head, neck, and tail size,
acrosome size, and evaluating for the presence of
vacuoles
Kruger’s criteria
95
REPORTING FOR KRUGERS CRITERIA
Normal: >14% in normal forms
96
Tests FOR SPERM MORPHOLOGY include
a. Sperm vitality
b. Seminal fluid fructose level
c. Sperm agglutinins
d. Microbial & Chemistry Tests
97
Normal sperm concentration but with decreased motility
sperm vitality
98
Method: sperm vitality
Semen + eosin- negrosin stain
✓ Count dead cells per 100 spermatozoa
99
Principle of sperm vitality
living cells cannot be penetrated by the stain
100
✓ Living cells stain
bluish white
101
✓ Dead cells stain
red against a purple background
102
✓ Normal:
50% or more living spermatozoa with 2.0 motility
103
Presence of vital but immotile cells =
defective
flagellum
104
large numbers of dead cells =
epididymal pathology
105
Low or absent fructose =
= low sperm concentration
106
Screening test for seminal fructose
Resorcinol test
107
(+) fructose =
orange-red color
108
Quantitative test for seminal fructose
Spectrophotometry
109
Normal spectophotometry
>13 micromole per ejaculate
110
Fructose analysis must be performed within
2 hours
or frozen specimen to avoid fructolysis
111
May be present in females and male (frequently encountered)
anti-sperm antibodies
112
➔ Disruption of blood-testes
barrier
➔ Sperm antigens trigger
immune system to produce
antibodies
➔ Occur after surgery,
vasectomy reversal
(vasovasostomy), trauma,
and infection
➔ Presence of clumps of
sperm during semenalysis
males
113
➔ Damaged sperm can cause
production of antibodies
➔ Infertility but normal
semenalysis findings
females
114
➔ Screening test to detect IgG
➔ Semen + AHG + latex particles or RBC treated with
IgG
Mixed agglutination reaction (MAR)
115
(MAR) Presence of antibody =
= agglutination of sperm and
particles/cell
116
Normal IN MAR
<10% of the sperm attached to the
particle/cell
117
➔ Specific test that detect:
✓ IgG, IgM, or IgA
✓ Area where antibody is located
Semen + polyacrylamide beads coated with anti-IgG,
anti-IgM, or anti-IgA
Immunobead Test
118
= interfere with penetration
into the cervical mucosa or ovum
Head-directed antibodies
119
= movement through the
cervical mucosa
Tail-directed antibodies
120
Presence of antibody = (IMMUNOBEAD)
beads attached to sperm at
particular areas
121
Normal: (IMMUNOBEAD)
Presence of beads on less than 50% of the
sperm
122
Reporting IN IMMUNOBEAD
Type of antibody and where it is located.
(For example: IgM tail antibodies,” “IgG head
antibodies)
123
➔ Aerobic and anaerobic cultures and tests for
Chlamydia trachomatis, Mycoplasma hominis, and
Ureaplasma urealyticum
● Microbial:
124
➔ Method: spectrophotometry
Chemistry:
125
✓ Decreased amounts indicate disorder of the
epididymis
Neutral "a-glucosidase, glycerophosphocholine, and
L-carnitine
126
✓ Decreased amounts indicate lack of prostatic
fluid
Zinc, citric acid, glutamyl transpeptidase, and acid
phosphatase
127
❖ Enzyme present in semen
❖ Detection can indicate presence of semen
Acid Phosphatase
128
❖ Presence of semen in the absence of
sperm
Prostatic Specific Antigen (PSA)
129
* Motile sperm can be detected up to
24 hours after intercourse
130
Non-motile sperm can persist up to
3 days
131
Head of the sperm can be seen
7 days after the intercourse
132
Method OF POST VASECTOMY
➔ Wet preparation
➔ If wet preparation is negative:
✓ Centrifugation of specimen for 10 minutes and
sediment is examined for spermatozoa
133
Presence of single “motile” sperm on a wet preparation =
unsuccessful vasectomy
134
Sperm are incubated with species-nonspecific
hamster eggs, and penetration is observed
microscopically
Hamster egg
penetration
135
Observation of sperm’s ability to penetrate
partner’s mid cycle cervical mucus
Cervical mucus
penetration
136
Sperm exposed to low-sodium concentrations
are evaluated for membrane integrity and sperm
viability
Hypo-osmotic
swelling
137
Evaluation of the acrosome to produce enzymes
essential for ovum penetration
In vitro acrosome
reaction
138
Determines the sperm velocity, trajectory,
concentration and morphology
Computer Assisted Semen Analysis (CASA) systems
139
Three CASA instrumentations:
a. Sperm Class Analyzer (SCA)
b. CEROS CASA systems
c. Automated Sperm Quality Analyzers (ASQA)
140
Microscope equipped with a digital camera,
motorized heating stage, and image analysis
software with analytical filter
Sperm Class Analyzer (SCA)
