Semen analysis is performed to evoluvate male firtility and reproductive health assessing the quality and quantity of sperm in a semen sample. Semen examination is an inexpensive test used for the infertility ,success of vasectomy and medicolegal  cases,hormonal imbalances, infections, lifestyle factors, or exposure to certain medications and environmental toxins.

semen analysis consist of following steps.

1.Rutine analysis:-

Routine Analysis of Semen is a laboratory test performed to evaluate the physical, microscopic, and functional characteristics of semen. The examination includes assessment of semen volume, color, viscosity, liquefaction time, pH, sperm concentration (count), motility, morphology , and the presence of other cells such as white blood cells or immature germ cells. process included:-

1.Sample collection

2.physical examination 

3.Chemical examination

4.Microscopic examination 

1.Sample collections:- 

A semen specimen should be collected after 3-7 days  of sexual absentees for accurate results.The sample is usually obtained by masturbation directly, the specimen is collected in a clean, sterile, wide-mouth ,leak-proof container provided by the laboratory or properly washed dry condom.then specimen submitted in a laboratory.

After collection, the specimen container properly labeled with the patient,s name ,date ,and time of collection. sample should be allowed to liquefy at room tempreture.

2.Physical examination or Gross examination:-

The physical examination of semen is the first step in routine semen analysis.its consist following aspect.

1.Volume:- Volume in a single ejaculation approximately 2.5-5 ml semen passed ,the volume is slightly more in patient of infertility. The volume dose not various with the periods of absenties in increase volume of semen is known as hyperspermia and low  volume is caused hypospermia.

2.colour:-Normally it is whitish ,gray white of slightly yellowish or milky.Yellowish or reddish discoloration may indicate infection or blood contamination.

3.pH(power of hydrogen ):- The normal pH  is 7-8.it is slightly alkaline in nature.

4.Viscosity :- During the ejaculation semen are viscus ,when ejaculation falls drop by drop .

5.Liquification:- Liquification occur because of presence of fibrinogen ,normally liquification occur at room tempreature within 10-30 minute avarage 20 minutes.

2. Chemical examination:-

Fructose test of semen:-

Fructose test is used to determined androgen deficiency or ejaculatory obstruction to semen .the level of seminal fructose is low in both condition fructose is measured qualitative by resorcinol’s.

The fructose test is performed to detect the presence and amount of fructose in seminal fluid. Fructose is produced mainly by seminal vesicles and serves as the primary energy source for sperm motility.

Requirements:- 

*Diluted HCL

*Resorcinol

*Semen/specimen

*Test tube 

*Bansal burner 

Procedure:-

1.  Take a 5 ml diluted HCL in a test tube.

2.  Add 1ml semen.

3. Add 5 mg resorcinol’s boil it for few minuts.

Normal value:-                                        150-600 mg/dl.

Interpretation:-

Appearence of red colour indicate the presence of fructose. it can measured by spectrophotometer.

                            -:Microscopic examination:-

Motility:- Motility with the help of liquefied semen make a wet mount sample slide and examin under the microscope first low power field and then highpowerr field microscope .normally within 2 hours of ejaculation mote than >60% spermatozoa  vagrasly motile, and in 6-8 hours 25-40% are motile.

                               -:Spermatozoa counting:-

1.Dilute the semen specimen in 1:10.

2.Charge the new baur’s chamber and weight for 2 minutes to allow the spermatozoa settle down.

3.Examine under microscope and count spermatozoa in 4 square cornor of new baur’s chamber and calculate them as follow.

Calculation :-  

Spermatozoa =  N×D/V

=N×10/0.4

=N×25

Spermatozoa /ml= N×25×1000

=N×25000

Normal values:-    

>60 million/ml

>60 × 10⁶/ml

Abnormal values:-

<20 × 10⁶/ml

Marphology examination of spermatozoa:-

For the morphology’s  study of spermatozoa prpared a thin smear from liquified semen on a glass slide,and stained it with any of the romanowashky stain,pap stain or H&E stain.

Observe atleast 200 spermatozoa for any abnormalities in their morphology normally 80% spermatozoa are normal.

Normal marphology of spermatozoa:-Abnormal marphology of specimen.

Abnormal marphology of specimen:-