Semen Analysis Flashcards
Inability to conceive after one year of unprotected intercourse or the inability to carry pregnancy to term
Infertility
Located in the interstitium of the testes and are responsible for the production of testosterone
Leydig cells
Area b/w the seminiferous tubules; not part of the seminiferous tubules
Interstitium
Accessory glands that are responsible for approximately 70% of the ejaculate; contains high concentrations of flavin and fructose
Seminal vesicles
Accessory gland that is responsible for approximately 25% of the ejaculate; contains acid phosphatase, proteolytic enzymes, and zinc
Prostate gland
Secretes an alkaline mucus that serves to buffer the ejaculate to a slightly alkaline pH
Bulbourethral glands
List the order of the travel route of sperm (SEVEn UP)
- Seminiferous tubules
- Epididymus
- Vans deferns
- Ejaculatory duct
- Urethra
- Penis
Proper collection of seminal fluid
- Collected through masturbation
- Collect in a warm glass container
- Store at 37°C until testing begins
- Collect b/w 2-7 days of abstinence
- Label w/ name, period of abstinence, and date/time of collection
- Testing should begin w/in 1 hour of collection
Normal appearance of seminal fluid
- Viscous
- Translucent gray/white
Abnormal appearance of seminal fluid
- “Whiter” or more turbid = leukocytes
- Red = expect RBCs
- Yellow = urine, prolonged abstinence, certain meds
Normal viscosity of seminal fluid
Semen immediately coagulates after ejaculation and w/in 30 minutes the coagulum will liquify
Abnormal types of viscosity
- Specimen isn’t liquefied by 60 minutes
- If clumps or agglutination persists, possibly due to Abs
- Watery samples due to few (if any sperm)
- Mucus threads form that are greater than 2 cm
Normal volume for seminal fluid
2-5 mL
What does a volume of seminal fluid < 2 mL indicate?
- Blockage
- Infection
- Congenital absence of seminal vesicles
- Retrograde ejaculation
- Loss of fluid
Normal pH for seminal fluid
7.2-8.0, slightly alkaline to neutralize acidic environment of vagina
What does a high pH of seminal fluid indicate?
Infections
What does a low pH of seminal fluid indicate?
A larger percentage of ejaculate volume is from the prostate gland or there are abnormalities in other sites
One of the most important factors for infertility testing, done on brightfield
Motility
How do you examine motility?
- Prepare a wet mount and count percentage of sperm in each graded category (rapid progressive, moderately progressive, non-progressive, and immotile)
- A sperm moves by rotating its tail in a spiral motion
Normal sperm motility
≥50% in the two most motile groups if the specimen is evaluated w/in 60 minutes
How do you analyze sperm concentration?
- Dilute specimen
- Load hematocytometer
- Count 4 corner squares and red cell counting area
- Count both sides and average
Equation for calculating sperm concentration
((avg # sperm)/(L x W x D)) x dilution factor
Normal concentration for sperm
200-250 million/mL, since sperm are reported in mL and no uL, you have to multiply by 1000
Reasons for low sperm concentration
- Anatomical defects
- Short periods of abstinence
- Loss of first portion of ejaculate
- Low or absent fructose
How do you calculate sperm count?
[sperm] x volume of ejaculate
Normal sperm count
≥ 40 million
When and why is a sperm count performed?
- After postvasectomy, specimens are collected at monthly intervals beginning 2 months after surgery
- Must have 2 monthly specimens w/ no sperm
- Presence of motile sperm indicates unsuccessful procedure
- Macroscopic results should still be normal
How do you analyze morphology?
- Prepare slide and air dry
- Stain w/ Wright’s Giemsa, or Papanicolaou
- Rate under oil immersion (1000x), count defective sperm and type of defect
Normal percentage of normal sperm morphology
12-15% or greater
What percentage of normal sperm indicates a strong predictor of infertility?
< 5% normal sperm
What is the teratozoospermia index (TZI)?
- Number of defects divided by a total number of abnormal sperm
- Should be ≤1.6, helpful in predicting sperm function
What would be the cause of abnormal morphology
- Anatomical defects
- Physiological problems
- Infection
- Scrotal heating
- Frequent ejaculations
Two stains useful in examining vitality
Blom’s Stain or eosin nigrosin will allow rapid differentiation between alive and dead sperm, where dead sperm take up stain and appear dark pink and live sperm do not take up stain and appear white
Normal vitality
≥50% are more alive in normal men
What do leukocytes in ejaculate indicate?
Should be < 1 million/uL of seminal fluid otherwise indicate infection or inflammation
What is measured in rape investigations?
- Look for motile sperm (24 hours) or immotile sperm (3 days)
- Acid phosphatase measurement
- PSA
______ is the only body fluid with high concentration of acid phosphatase and can be measured in vaginal fluid or from skin or clothing
Semen
Semen is routinely analyzed to evaluate what?
Post-vasectomy status
Following liquefaction, what should the viscosity of the ejaculate look like?
Viscosity is similar to water
A semen pH > 8.0 is associate with what?
An infection in the male reproductive tract
Vitality staining with these two stains enables rapid differentiation b/w alive and dead sperm
Blom’s and eosin nigrosin
- Dead sperm take up the stain and appear pink, live sperm appear white
- Serves as a confirmation for motility
What else could be found in the ejaculate?
- Leukocytes (< 1 million/mL; if higher, indicates infection or inflammation)
- Sperm precursors
- Urethral epi cells
- Cellular debris
Normal of combined leukocytes and sperm precursors (round cells) in seminal fluid
≤ 5 million/mL
Two tests that test for Abs against sperm
- Mixed agglutination rxn
- Immunobead test
Mixed agglutination
- Principle
AHG is mixed w/ latex beads coated w/ IgG. AGH binds to latex beads and sperm simultaneously in the presence of a sperm Abs
Immunobead test
- Principle
Sperm are added to a solution containing latex beads that are coated w/ anti-IgG (anti-IgM or anti-IgA). Microscopic analysis indicates whether the Abs are directed against the head or the tail
