Semen analysis Spermogram

1
Semen analysis(Spermogram)
2

• Collected in
• - a glass or plastic jar free of
  any
• preservative
• -warm ( at least to room T. )
• Evaluated
• within 30 min after collection
• (no examination after 2 hrs)

3
Record

• 1- The name of the man
• 2- The period of abstinence
• 3- The date time of collection
• 4- Completeness of collection
• 5- Difficulties in producing the sample
• 6- The interval between collection analysis

4
Two samples should be collected for initial
evaluation( interval 7 days 3 weeks )
5
Macroscopic analysis

• 1- PH (measured with PH paper)
• 2-Volume (measured by weight or using
• graduated cylinder)
• 3-Appearance abnormal smell ,color
• or viscosity( thread shorter
  than 2 cm)
• 4-Liquefaction time (above 60 min )
• bromelain ?
• 5-Seeding for bacterial analysis

6
Microscopic analysis

• 1-Sperm concentration
• (total number , viability sp.
  agglutination)
• - Centrifuge if indicated
• - Examine at least 200 sp. For motility
  
• 2- Presence of other cellular elements
  

7
Centrifugation properties

• 1- sp. lt1-2 / field 1 ml at 600 g for
• 15 min
• 2-no sp. 1 ml at 3000 g for
• 15 min

8
Preparation for routine analysis

• A fixed volume of 10 ul semen onto a clean
• glass slide covered with a 22mm / 22mm
• coverslip
• Evaluation after one minute ( for stabilization )
• Assessment at 37 C ( or a constant temperature
• between 20 40 C )

9
Grading system for sperm motility

• a. Rapid progressive motility
• (gt25 um/s at 37 C and gt20 um/s at 20 C)
• b. Slow or sluggish progressive motility
• c. Nonprogressive motility (lt5 um/s )
• d. Immotility

10
Preliminary estimation of sperm concentraton

• This is used to decide the dilution for
  determining
• the sperm concentration by haemacytometer.
• - Counting spermatozoa per 400 field
• - Preparing appropriate diluent
• (distilled water NaHCO3 formalin gentian
• violet or trypan blue )
• - Dilute the semen based on estimated sperm
• concentration

11
Assessment of sperm concentration

• 1 - It should be determined using the
• haemocytometer method
• - 10 ul of the thoroughly mixed diluted specimen
  is transferred to the counting chamber
• - Counting spermatozoa in the central square of
  the
• grid (25 large squares ) , the number of
  large
• squares are determined based on the number of
  sp.
• per each square ( to 200 sp. )
• - Calculate total number by using the formula.

12
Assessment of sperm concentration

• 2 alternative method
• - 1 ml well-mixed sample 19 ml cold water
• - Fill a hematology counting chamber
• - Count number of sperm seen in a 1 mm2
  area
• ( 16 small squares ) . This number is then
• multiplied by 200,000 )
• Double figure and add 5 zeros
  !!

13
Limitations of the porocedure

• Delayed examination of the specimen
• Collection in improper container
• Exposure of the specimen to temperature extremes
• during transport
• Abnormally low sperm count allowing for
• evaluation of less than 200 spermatozoa
• Use of dirty or contaminated supplies

14
Semen culture

• Abstinence for 5-7 days
• Passing urine before obtaining the sample
• C leang the genitalia with soap
• Collect the specimen in a sterile container
• Examine the specmen within 3 hrs after collection
• Culture in a G and G- medium