LAB 01 - SEMEN ANALYSIS Flashcards
secretion of sperm
Testes (seminiferous tubules
sperm maturation and development of flagella; stores sperms
Epididymis
transports sperm to the ejaculatory duct
Ductus deferens (vas deferens)
transport medium for the sperm; rich in fructose and flavin.
Seminal vesicle
propels the sperm through the urethra; contains acid phosphatase, citric acid, zinc, and proteolytic enzymes responsible for both the coagulation and liquefaction of the semen
Prostate gland
secretes alkaline mucus.
Bulbourethral glands
COMPOSITION OF SEMEN
5% WHAT, DUHA TO SILA!
Spermatozoa AND Bulbourethral fluid
COMPOSITION OF SEMEN
__% Seminal Fluid
60-70%
COMPOSITION OF SEMEN
__% Prostate Fluid
COMPOSITION OF SEMEN
thick alkaline mucus
Bulbourethral fluid
PURPOSE
GIVE ME THOSE 3 UNDER THIS
- Evaluation of reproductive dysfunction in males— infertility testing
- Select donors for therapeutic insemination
- Monitor success of surgical procedures:
varicocelectomy
Vasectomy
SAMPLE COLLECTION
PREPARATION:
- Abstinence: 2-7 days (Note: No prolonged abstinence)
- Sterile glass or plastic container
* Collected in a private room near the laboratory Give a clear written and verbal instructions concerning the collection of semen
* Deliver to the laboratory within 1 hour after collection
* Record the patient’s name, the period of sexual abstinence, and the date and time of specimen collection
METHODS OF COLLECTION
GIVE ME THOSE 4 UNDER THIS
- Masturbation
- Coitus interruptus (only in exceptional cases)
- Silastic seminal fluid collection device
- Aspiration from the vaginal vault after coitus
APPEARANCE:COLOR
NORMAL : ?
Gray-white, “Pearly-white”, Light Yellow; Opaque
APPEARANCE:COLOR
Shades of yellow : __
Correlate with flavin concentration
APPEARANCE:COLOR
__ : Associated with certain drugs, over abstinence, jaundice, urine contamination
Deep yellow
APPEARANCE:COLOR
Brown or red: ?
Presence of red blood cells
APPEARANCE:COLOR
__ : Presence of white blood cells and infection within the reproductive tract
Increased white turbidity
After ejaculation, semen is typically a semi-solid coagulated mass as protection from the acidic vaginal pH. Liquefies within ___ after collection (Normal)
30 to 60 minutes
Failure of liquefaction within 60 minutes -
eficiency in prostatic enzymes and should be reported.
If after 2 hours the specimen has not liquified:
- Dulbecco’s phosphate -buffered saline
- Proteolytic enzymes such as alpha - chymotrypsin or bromelain
Normal volume: __
2-5 mL
High volume MEANS
-active exudation in cases of active inflammation of the accessory organs -extended abstinence
LOW VOLUME MEANS
-obstruction of the ejaculatory duct, congenital bilateral absence of vas deferens, collection problems (loss of fraction of the ejaculate), partial retrograde ejaculation or androgen deficiency
Normal viscosity:
formation of small discrete droplets that do not appear like a clump or stringy when falling by gravity
Abnormal viscosity: __
drop from a thread more than 2 cm long (viscous)
pH paper with a range of ___ should be used
4.0 to 10.0
Normal pH: ___
7.2 – 7.8
Increased pH:
infection
Decreased pH:
increased prostatic fluid, ejaculatory duct obstruction, poorly developed seminal vesicle
SPERM MOTILITY
assessed by checking sperm cells in __ HPFs or 200 sperms in 2 replicates
20
The lower reference limit for total motility (PR+NP) is __
40%.
The lower reference limit for progressive motility (PR) is
32%.
SPERM MORPHOLOGY
GIVE ME THE 5 BASIC PARTS
- HEAD- oval-shaped; approx. 5 um long and 3 um wide
- TAIL- approx. 45 um long (with axoneme)
- ACROSOMAL CAP- encompasses half of the head; cover 2/3 of sperm nucleus
- MIDPIECE- 7 um long; mitochondrial sheath
- MOTILITY: 1-4 mm/min; Life span: 1-2 days at body temp
Normal values (count at least ___ sperm cells)
200
Kruger’s Strict Criteria: >__% normal; measures head, neck and tail using a micrometer
14%
Routine criteria: >__% normal
30%
number of spermatozoa per unit volume of semen
Sperm concentration
total number of spermatozoa in the entire ejaculate and is obtained by multiplying the sperm
Total sperm number/Sperm count
Normal values: >__ million/ejaculate
Normal values:__ million sperm per milliliter
(1) 40
(2) 20-250
Improved Neubauer Counting Chamber
- __:__ dilution
1:20
Diluting fluids: GIVE ME THE 5 WITH PERCENT AND TULO DAPAT
cold water, formalin, Na bicarbonate, 0.5% chlorazene, 1% formalin in 3% trisodium citrate
5 RBC squares - Computation:
sperm cells counted x 1,000,000 (mL)
2 WBC - Computation:
sperm cells counted x 100,000 (mL)
Addition of __ (crystal violet) to diluting fluid helps visualization using bright-field microscope.
stain
> 1 million WBCs/mL - ___
inflammation or infection
> 1 million spermatids/mL -
disruption of spermatogenesis caused by viral infections, exposure to toxic chemicals, and genetic disorders
no ejaculate at all (anejaculation or retrograde ejaculation)
Aspermia
less than 20 million per milliliter
Oligospermia
immotile/dead sperms
Necrospermia
complete absence of sperm
Azoospermia
OTHER TSTS AND TECHNIQUES
NAME ME THE 3
- SPERM VITALITY
- SEMINAL FLUID FRUCTOSE
- ANTISPERM ANTIBODIES
- Motile spermatozoa sticking to each other, head-to-head, tail-to-tail or in a mixed way.
- Motility is vigorous with frantic shaking motion or with limited motion
AGGLUTINATION OF SPERMATOZOA
Grade ___ - isolated - <10 spermatozoa per agglutinate, many free spermatozoa
1
Grade 2 - moderate - ___ spermatozoa per agglutinate, free spermatozoa
10-50
Grade __ - gross - all spermatozoa agglutinated and agglutinates interconnected
4
Grade __ - large - agglutinates of >50 spermatozoa, some spermatozoa still free
3
-screening procedure to detect IgG antibodies
Mixed Agglutination Reaction (MAR test)
Mixed Agglutination Reaction (MAR test)
Normal: < __% of motile sperm attached to particles
10
more specific to detect IgG, IgM, and IgA antibodies and demonstrates what area of sperm (head, neckpiece, midpiece, or tail) the autoantibodies are affecting.
Immunobead test
Normal: presence of beads on <__% of the sperm
50
> __ million WBCs/ mL indicates infection within reproductive system, prostate.
1
CHEMICAL TESTING
Decreased neutral alpha-glucosidase, glycerophosphocholine and L-carnitine suggest a disorder of ___.
epididymis
CHEMICAL TESTING
Decreased zinc, citric acid, glutamyl transpeptidase and acid phosphatase indicate lack of ____.
prostatic fluid
SPECIAL CASES
In cases of alleged rape -microscopic examination of the specimen for the presence of sperm with __
xylene
T OR F
SPECIAL CASES
Motile sperm- 3 days
Nonmotile sperm- detected for up to 24 hours after intercourse
F
Motile sperm- detected for up to 24 hours after intercourse
Nonmotile sperm- 3 days
SPECIAL CASES
__ - more specific detection
Seminal glycoprotein p30
POST VASECTOMY ANALYSIS
concerned only in presence or absence of __
spermatozoa
T OR F
POST VASECTOMY ANALYSIS
Dry preparation using phase microscopy
F - WET
- Performed in specialized andrology laboratories
SPERM FUNCTION TEST
SPERM ARE INCUBATED WITH SPECIES-NONSPECIFIC HAMSTER EGGS AND PENETRATION IS OBSERVED MICROSCOPICALLY
Hamster egg penetration assay
OBSERVATION OR SPERM’S ABILITY TO PENETRATE PARTNER’S MIDCYCLE CERVICAL MUCUS
Cervical mucus penetration test
EVALUATION OF THE ACROSOME TO PRODUCE ENZYMES ESSENTIAL FOR OVUM PENETRATION
In-vitro acrosome reaction
SPERM EXPOSED TO LOW-SODIUM CONCENTRATIONS ARE EVALUATED TO MEMBRANE INTEGRITY AND SPERM VIABILITY
Hypo-osmotic swelling test
BALIK PARA 75 TANAN
In cases of alleged rape -microscopic examination of the specimen for the presence of sperm with __
xylene