F1: Semen Analysis (Part 3: Macroscopic and Microscopic Examination) Flashcards
MACROSCOPIC EXAMINATION
Macroscopic examination involves 5 parameters, what are those?
- Appearance/ Color
- Volume
- Liquefaction Time
- Viscosity
- pH
VALVP
APPEARANCE/COLOR
Identify color based on description of appearance:
Gray white/pearly white translucent w/ musty odor or chlorox like smell
MACROSCOPIC EXAMINATION
Normal
APPEARANCE/COLOR
Identify color based on description of appearance:
WBCs which indicate infection
MACROSCOPIC EXAMINATION
Turbid
APPEARANCE/COLOR
Identify color based on description of appearance:
indicate low sperm count
MACROSCOPIC EXAMINATION
Clear
APPEARANCE/COLOR
Identify color based on description of appearance:
indicate urine contamination, prolonged abstinence, medications
MACROSCOPIC EXAMINATION
Yellow
APPEARANCE/COLOR
Identify color based on description of appearance:
Flavin
MACROSCOPIC EXAMINATION
Gray
APPEARANCE/COLOR
Identify color based on description of appearance:
RBCs
MACROSCOPIC EXAMINATION
red
APPEARANCE/COLOR
Yellow color of sperm
a. Urine contamination
b. Prolonged abstinence
c. Medication
d. ALL
e. a and b
f. b and c
g. nOTA
MACROSCOPIC EXAMINATION
d. ALL
VOLUME
Semen is measured in what container and how many mL of increments?
MACROSCOPIC EXAMINATION
container: clean graduated cylinder in 0.1 mL increments
VOLUME
Identify if Normal, Low volume, High volume
2-5 mL per ejaculate
MACROSCOPIC EXAMINATION
Normal
VOLUME
Identify if Normal, Low volume, High volume
- Non-adherence to 2-day abstinence
- Infertility
- Problem in the seminal vesicle
MACROSCOPIC EXAMINATION
Low volume
VOLUME
Identify if Normal, Low volume, High volume
Signifies prolonged abstinence
MACROSCOPIC EXAMINATION
HIgh volume
LIQUEFACTION TIME
Identify if Normal liquefaction time or Prolonged liquefaction time
30 – 60 minutes after collection
MACROSCOPIC EXAMINATION
Normal liquefaction time
LIQUEFACTION TIME
Identify if Normal liquefaction time or Prolonged liquefaction time
Indicates deficiency in prostatic enzymes
MACROSCOPIC EXAMINATION
Prolonged liquefaction time
LIQUEFACTION TIME
T or F
SPecimen should be analyzed before liquefaction
MACROSCOPIC EXAMINATION
F (should be analyzed after liquefaction)
LIQUEFACTION TIME
If after 2 hours specimen has not liquefied, what (2) reagents should be added?
MACROSCOPIC EXAMINATION
- Dulbecco’s phosphate-buffered saline
- Proteolytic enzymes (alphachymotrypsin bromelain)
LIQUEFACTION TIME
T or F
Liquefaction should be timed
MACROSCOPIC EXAMINATION
T
VISCOSITY
T or F
To observe viscosity, pour the specimen and observe how it pours OR by using pasteur pipette
MACROSCOPIC EXAMINATION
T
VISCOSITY
Identify if Normal or Highly viscous
Pours like droplets
MACROSCOPIC EXAMINATION
Normal
VISCOSITY
Identify if Normal or Highly viscous
Droplets with threads of >2cm
MACROSCOPIC EXAMINATION
Highly viscous
VISCOUS
Rating of 0 or 4 ?
Watery
MACROSCOPIC EXAMINATION
0
VISCOUS
Rating of 0 or 4 ?
Gel-like
MACROSCOPIC EXAMINATION
4
VISCOUS
T or F
Viscosity can also be reported as low, normal, or high viscosity
MACROSCOPIC EXAMINATION
T
VISCOUS
Highly clumped and viscous semen indicates what motility ?
MACROSCOPIC EXAMINATION
slower sperm motility
pH
pH of semen can be detected by what (2) methods?
MACROSCOPIC EXAMINATION
- pH pad of urinalysis reagent strip
- pH paper
pH
Identify if High pH, Normal pH, Low pH
indicates infection (reproductive tract)
MACROSCOPIC EXAMINATION
High pH
pH
Identify if High pH, Normal pH, Low pH
7.2 – 8.0
MACROSCOPIC EXAMINATION
Normal
pH
Identify if High pH, Normal pH, Low pH
- Increased prostatic fluid
- Obstruction of the ejaculatory duct
- Problem in the seminal vesicle
MACROSCOPIC EXAMINATION
Low pH
MICROSCOPIC EXAMINATION
Microscopic examination involves 3 parameters, what are those?
- Motility
- Morphology
- Count
SPERM CONCENTRATION/COUNT
Identify if Normal sperm concentration, Borderline concentration, Normal sperm count:
20-250 Million sperm/mL
MICROSCOPIC EXAMINATION
Normal sperm concentration
SPERM CONCENTRATION/COUNT
Identify if Normal sperm concentration, Borderline concentration, Normal sperm count:
10-20 Million/mL
MICROSCOPIC EXAMINATION
Borderline concentration
SPERM CONCENTRATION/COUNT
Identify if Normal sperm concentration, Borderline concentration, Normal sperm count:
At least 40 Million/ejaculate
MICROSCOPIC EXAMINATION
Normal sperm count
SPERM CONCENTRATION/COUNT
Formula of sperm count?
MICROSCOPIC EXAMINATION
Concentration x Volume
SPERM CONCENTRATION/COUNT
Common dilution is?
MICROSCOPIC EXAMINATION
1:20
SPERM CONCENTRATION/COUNT
Common dilution of 1:20 is achieved using what equipment?
MICROSCOPIC EXAMINATION
tatlo
Mechanical (positive-displacement) pipette/Automatic pipette/ WBC pipette
Pwede RBC pipette pero mahirap gamitin dahil white(semen) to white(background), mahihirapan basahin
SPERM CONCENTRATION/COUNT
T or F
Dilution is done to mobilize sperm prior to counting
MICROSCOPIC EXAMINATION
F (dilution is to immobilize sperm before counting)
SPERM CONCENTRATION/COUNT
What are the (3) diluting fluids used?
MICROSCOPIC EXAMINATION
- Sodium bicarbonate
- Formalin
- Chilled water
- Saline
SPERM CONCENTRATION/COUNT
Most common which dissolves mucus
MICROSCOPIC EXAMINATION
(3) diluting fluids used
Sodium bicarbonate
SPERM CONCENTRATION/COUNT
Most common which immobilizes sperm
MICROSCOPIC EXAMINATION
(3) diluting fluids used
Formalin
SPERM CONCENTRATION/COUNT
Used to kill sperm to stop motility
MICROSCOPIC EXAMINATION
(3) diluting fluids used
Chilled water
SPERM CONCENTRATION/COUNT
- Used in counting sperm cells
- Only the mature cells are counted, not the immature “round cells”
MICROSCOPIC EXAMINATION
Neubauer chamber
SPERM CONCENTRATION/COUNT: Neubauer chamber
T or F
In neubauer chamber, count the round cells and not the cells with head or tail
MICROSCOPIC EXAMINATION
F (reverse; count cells with head or tail or MATURE CELLS, NOT the round cells or IMMATURE CELLS)
SPERM CONCENTRATION/COUNT
This defines the square’s boundary (black line, left panel)
MICROSCOPIC EXAMINATION
middle of the 3 lines
SPERM CONCENTRATION/COUNT: Neubauer chamber
T or F
All spermatozoa within the central square are counted, as well as those with their heads between the 2 inner lines (white circles), including whose heads lie between the outer 2 lines (black circles)
MICROSCOPIC EXAMINATION
F (All spermatozoa within the central square are counted, as well as those with their heads between the 2 inner lines (white circles), but not those whose heads lie between the outer 2 lines (black circles))
SPERM CONCENTRATION/COUNT: Neubauer chamber
is counted only if that line is the lower or left-hand line of the square (white circle, middle panel) but not if it is the upper or right hand line of the square
MICROSCOPIC EXAMINATION
A spermatozoon with most of its head lying on the central line
SPERM CONCENTRATION/COUNT: Neubauer chamber
Sperm concentration per mL formulation
Using 2 WBC squares
MICROSCOPIC EXAMINATION
Sperm concentration per mL =
no. of cells counted x DF (20) x 1,000 / no. of squares (2) x volume of 1 square (0.1)
OR
no. of cells counted x DF (20) x 1,000 / no. of squares (2)
no. of cells x 20 x 1,000 / 2 x 0.1
MICROSCOPIC EXAMINATION
SPERM CONCENTRATION/COUNT: Neubauer chamber
Simplified concentration per mL formula
Using 2 WBC squares
MICROSCOPIC EXAMINATION
no. of cells counted x 100,000
SPERM CONCENTRATION/COUNT: Neubauer chamber
T or F
When using 5 RBC squares one should load one side of hemocytometer
Using 5 RBC squares
MICROSCOPIC EXAMINATION
f (Load both sides of hemocytometer)
SPERM CONCENTRATION/COUNT: Neubauer chamber
After loading both sides, sample should be allowed to settle for how many minutes?
MICROSCOPIC EXAMINATION
3 to 5 minutes
SPERM CONCENTRATION/COUNT: Neubauer chamber
After settling, two counts should be?
MICROSCOPIC EXAMINATION
Averaged
SPERM CONCENTRATION/COUNT: Neubauer chamber
T or F
Average of 2 counts should agree within 10%
MICROSCOPIC EXAMINATION
T
SPERM CONCENTRATION/COUNT: Neubauer chamber
If average of 2 counts do not agree, what should one do?
MICROSCOPIC EXAMINATION
repeat dilution and counting multiply the counted
sperm by 1,000,00
SPERM CONCENTRATION/COUNT: Neubauer chamber
Sperm concentration per mL formulation
Using 5 RBC squares
MICROSCOPIC EXAMINATION
Sperm concentration per mL = no. of cells counted x DF (20) x 1,000 / no. of sqaures (5) x volume of 1 square (0.004)
no. of cells x 20 x 1,000 / 5 x 0.004
SPERM CONCENTRATION/COUNT: Neubauer chamber
Simplified concentration per mL formula
Using 5 RBC squares
MICROSCOPIC EXAMINATION
no. of cells counted x 1,000,000
SPERM CONCENTRATION/COUNT: Neubauer chamber
- for counting sperm cells
- Uses a cover plate with 1mm2 grid divided into 100 squares
- undiluted specimen
- No additional factors are necessary for calculation
- number of spermatozoa counted in any strip of 10 squares of the grid indicates their concentration in
millions/mL
MICROSCOPIC EXAMINATION
Makler counting chamber
SPERM CONCENTRATION/COUNT
Makler counting chamber
a. cover plate with 1mm grid divided into 1000 squares
b. diluted specimen
c. additional factors
d. ALL
e. NOTA
MICROSCOPIC EXAMINATION
e. NOTA
a. cover plate with 1mm2 grid divided into 100 squares
b. **undiluted **specimens
c. NO additional factors
SPERM CONCENTRATION/COUNT
How are number of spermatozoa counted?
MICROSCOPIC EXAMINATION
MAKLER COUNTING CHAMBER
any strip of 10
squares of the grid
concentration in millions/mL
SPERM CONCENTRATION/COUNT
Causes of low sperm count could be because of (3) factors, what are those?
MICROSCOPIC EXAMINATION
MAKLER COUNTING CHAMBER
Medical causes, Environmental causes, Health and lifestyle and other causes
SPERM CONCENTRATION/COUNT
Medical, Environmental, Health/Lifestyle/other causes?
- Varicocele
- Ejaculation problems
- Anti-sperm Antibodies
- Tumors
- Undescended testicles
- Defects of tubules that transport sperm
- Chromosome defects
- Certain medications
- Prior surgeries
MICROSCOPIC EXAMINATION
MAKLER COUNTING CHAMBER
Medical causes
SPERM CONCENTRATION/COUNT
Medical, Environmental, Health/Lifestyle/other causes?
- Industrial chemicals
- Heavy metal exposure
- Radiation or X-rays
- Overheating the testicles
MICROSCOPIC EXAMINATION
MAKLER COUNTING CHAMBER
Environmental causes
SPERM CONCENTRATION/COUNT
Medical, Environmental, Health/Lifestyle/other causes?
- Drug use
- Alcohol use
- Tobacco smoking
- Emotional stress
- Weight
MICROSCOPIC EXAMINATION
MAKLER COUNTING CHAMBER
Health/Lifestyle/other causes
SPERM MOTILITY
T or F
Sperm must penetrate the cervical mucosa to the uterus, fallopian tubes and ovum
MICROSCOPIC EXAMINATION
MAKLER COUNTING CHAMBER
T
SPERM MOTILITY
Sperm motility can be detected using what (2) methods?
- Hanging drop method
- Sperm vitality
SPERM MOTILITY: 2 detection methods
- Undiluted, well mixed and fully-liquefied semen
- 10ul and 22 X 22 coverslip, concave slide
- % of motile sperm cells ( >50% within 1 hour)
MICROSCOPIC EXAMINATION
Hanging drop method
SPERM MOTILITY: 2 detection methods
volume of sample and what size of coverslip?
Hanging drop method
MICROSCOPIC EXAMINATION
sample volume: 10 uL
coverslip size: 22x22 (concave)
SPERM MOTILITY: 2 detection methods
Afer addition of samples, sample should be allowed to stand for how many minutes?
Hanging drop method
MICROSCOPIC EXAMINATION
1 minute
SPERM MOTILITY: 2 detection methods
Specimens detected through hanging drop method should be observed in how many high power fields?
Hanging drop method
MICROSCOPIC EXAMINATION
in 20 high power fields
SPERM MOTILITY: 2 detection methods
Normal % of motile sperm cells within 1hour?
Hanging drop method
MICROSCOPIC EXAMINATION
> 50% within 1 hour
SPERM MOTILITY: 2 detection methods
Identify Grade based on WHO criteria and Sperm motility
WHO Criteria: a
Sperm Motility: Rapid, straight line motility
Criteria for Hanging drop method
MICROSCOPIC EXAMINATION
4.0
SPERM MOTILITY: 2 detection methods
Identify Grade based on WHO criteria and Sperm motility
WHO Criteria: b
Sperm Motility: Slower speed, some lateral movement
Criteria for Hanging drop method
MICROSCOPIC EXAMINATION
3.0
SPERM MOTILITY: 2 detection methods
Identify Grade based on WHO criteria and Sperm motility
WHO Criteria: b
Sperm Motility: Slow forward progression, noticeable lateral movement
Criteria for Hanging drop method
MICROSCOPIC EXAMINATION
2.0
SPERM MOTILITY: 2 detection methods
Identify Grade based on WHO criteria and Sperm motility
WHO Criteria: c
Sperm Motility: No forward progression
Criteria for Hanging drop method
MICROSCOPIC EXAMINATION
1.0
SPERM MOTILITY: 2 detection methods
Identify Grade based on WHO criteria and Sperm motility
WHO Criteria: d
Sperm Motility: No movement
Criteria for Hanging drop method
MICROSCOPIC EXAMINATION
0
SPERM MOTILITY: 2 detection methods
Give interpretation based on result of sperm motility:
> 50% within 1 hour
Sperm Motility Interpretation hanging drop
MICROSCOPIC EXAMINATION
Normal
SPERM MOTILITY: 2 detection methods
Give interpretation based on result of sperm motility:
Motile in categories a, b, and c
Sperm Motility Interpretation hanging drop
MICROSCOPIC EXAMINATION
50% or more
SPERM MOTILITY: 2 detection methods
Give interpretation based on result of sperm motility:
show progressive motility (a and b)
Sperm Motility Interpretation Hanging drop
MICROSCOPIC EXAMINATION
25% or more
SPERM MOTILITY: 2 detection methods
T or F
One should observe immobile sperm and cliumps or antibodies when using hanging drop method
Sperm Motility Interpretation Hanging drop
MICROSCOPIC EXAMINATION
T
SPERM MOTILITY: 2 detection methods
If there is high percentage of immobile sperm, what test could be performed?
Sperm Motility Interpretation Hanging drop
MICROSCOPIC EXAMINATION
Sperm
vitality testing
SPERM MOTILITY: 2 detection methods
Clumping of sperm indicates possible presence of?
Sperm Motility Interpretation Hanging drop
MICROSCOPIC EXAMINATION
Sperm
agglutinins
SPERM MOTILITY: 2 detection methods
Progressive motility, Nonprogressive motility, Immotility?
Sperm moving linearly or in large circle
Alternative Sperm Grading Criteria OF Hanging drop
MICROSCOPIC EXAMINATION
Progressive motility
PM
SPERM MOTILITY: 2 detection methods
Progressive motility, Nonprogressive motility, Immotility?
Sperm moving in an absence of progression
Alternative Sperm Grading Criteria OF Hanging drop
MICROSCOPIC EXAMINATION
Nonprogressive motility
NP
SPERM MOTILITY: 2 detection methods
Progressive motility, Nonprogressive motility, Immotility?
No movement
Alternative Sperm Grading Criteria OF Hanging drop
MICROSCOPIC EXAMINATION
Immotility
IM
SPERM MOTILITY: 2 detection methods
Motility must be specified as?
Hanging drop
MICROSCOPIC EXAMINATION
Total Motility (PM and NP) or Progressive Motility (PM)
SPERM MOTILITY: 2 detection methods
Familiarize the parameters detected by Computer-Assisted Semen Analysis (CASA)
Hanging drop
MICROSCOPIC EXAMINATION
- Sperm velocity and trajectory – direction of motion
- Sperm concentration
- Sperm morphology
SPERM MOTILITY: 2 detection methods
- Performed using Bloom’s Test or Eosin/Nigrosin Test
- Done in cases of infertility where the sperm count is normal but has decreased motility
- Evaluate 100 sperm cells
Sperm vitality (viability)
SPERM MOTILITY: 2 detection methods
T or F
Sperm vitality or viability is perfromed in cases of infertility where sperm count is abnormal and have decreased motility
SPERM VITALITY (VIABILITY)
MICROSCOPIC EXAMINATION
F (NORMAL sperm count, but decreased motility)
SPERM MOTILITY: 2 detection methods
Interpret based on description of appearance:
* Not infiltrated by eosin
* Remain bluish white in color
SPERM VITALITY (VIABILITY)
MICROSCOPIC EXAMINATION
Living sperm cells
SPERM MOTILITY: 2 detection methods
Interpret based on description of appearance:
Red (stained by eosin)
SPERM VITALITY (VIABILITY)
MICROSCOPIC EXAMINATION
Dead sperm
SPERM VITALITY (VIABILITY)
SPERM MOTILITY: 2 detection methods
Interpret based on description of appearance:
Defective flagellum
SPERM VITALITY (VIABILITY)
MICROSCOPIC EXAMINATION
High number of vital but
immobile cells
SPERM MOTILITY: 2 detection methods
Interpret based on description of appearance:
Epididymal pathology
SPERM VITALITY (VIABILITY)
MICROSCOPIC EXAMINATION
High number of immotile
and nonviable cells
Sperm Morphology
Observe for head and tail morphology under what objective?
MICROSCOPIC EXAMINATION
Oil Immersion
Objective (OIO)
Sperm Morphology
How many sperms are evaluated?
MICROSCOPIC EXAMINATION
200 sperms
Sperm Morphology
T or F
In sperm morphology, one should take not of normal and abnormal forms
MICROSCOPIC EXAMINATION
T
Sperm Morphology
(3) stains used in sperm morphology?
MICROSCOPIC EXAMINATION
- Papanicolaou
- Giemsa
- Hematoxylin
PGH
Sperm Morphology: Preparation of normal semen smear
angle of slide when dragging spcimen?
MICROSCOPIC EXAMINATION
45 degrees
Sperm Morphology
What are the (3) distinct parts?
MICROSCOPIC EXAMINATION
- Head
- Middle piece
- Tail
Sperm Morphology
- For ovum penetration
- Oval shaped head approximately 5 µm long and 3 µm wide
- Acrosomal cap: approximately (40-70%) ½
(3) distinct parts
MICROSCOPIC EXAMINATION
Head
Sperm Morphology
T or F
Head is 3 um long and 5 um wide
(3) distinct parts
MICROSCOPIC EXAMINATION
F (head is 5 um long, 3um wide)
Sperm Morphology
Acrosomal cap occupies how big of the head?
(3) distinct parts
MICROSCOPIC EXAMINATION
½
of head (40-70%)
Sperm Morphology
- 7 um
- Contains mitochondria that provide energy for flagellar tail motion
(3) distinct parts
MICROSCOPIC EXAMINATION
Middle piece
Sperm Morphology
- For motility
- 40- 45 µm long
(3) distinct parts
MICROSCOPIC EXAMINATION
Tail
Sperm Morphology
Head or Tail abnormalities?
will result to poor ovum penetration
Abnormalities
MICROSCOPIC EXAMINATION
Head abnormalities
Sperm Morphology
Head or Tail abnormalities?
affect motility of the sperm
Abnormalities
MICROSCOPIC EXAMINATION
Tail abnormalities
Sperm Morphology
This abnormality causes the sperm head to bend backward and interfere with motility
Abnormalities
MICROSCOPIC EXAMINATION
Abnormally long neckpiece
Sperm Morphology
What are the (3) methods used for analysis of sperm morphology
- Kruger’s-strict criteria
- Computer-assisted semen analysis (CASA)
- SQA-V Sperm quality analyzer
Sperm Morphology
- Recommended by WHO but not routinely performed
- A system of evaluating sperm morphology using morphometry and stage micrometer
- Strictly measures the head, neck, tail, acrosomal head and
vacuoles - Normal forms (strict criteria): 14%
- Normal forms (routine criteria): 30%
(3) methods used for analysis of sperm morphology
MICROSCOPIC EXAMINATION
Kruger’s-strict criteria
Sperm Morphology
Tests for:
* Sperm velocity and trajectory (direction of motion)
* Sperm concentration
* Sperm morphology
(3) methods used for analysis of sperm morphology
MICROSCOPIC EXAMINATION
COMPUTER-ASSISTED SEMEN ANALYSIS (CASA)
Sperm Morphology
- Automatic results in 75 seconds
- Measures Motility (PM+NP)
- Immotility (IM)
- % Normal Morphology
- Average velocity (VELOCITY)
- Sperm Motility Index (SMI)
- Total sperm concentration (TSC)
- Motile sperm concentration (MSC)
- Progressively motile sperm
concentration (PMSC)
(3) methods used for analysis of sperm morphology
MICROSCOPIC EXAMINATION
SQA-V SPERM QUALITY ANALYZER