Friday, June 5, 2020

STUDY OF SEMEN

SEMEN ANALYSIS

Introduction

Problem with male semen a counts for 40% of all infertility another 40% released to female reproducts and harmon problem. The remaining 20% the pather semen Analysis is the cornerstone of testing for male in fertility this test provides important information about the quality and quantity of the sperm.

Semen is a composite solution consisting basically of spermatozoa suspended in the seminal plasma.

Semen is viscid neutral by slightly Alkaline and pale yellow colour due to its flavien content.

 

Collection of specime3n (Semen)

The usually recommended specimen one collected following a three day period of continence.

The most satisfactory – The specimen collected by masturbation in the clinical pathological Laboratory. This aloss a complete examination of the sperm particularly the liquification.

The specimen should be delivered with in 30min in laboratory.

(Note: - Wide mouth clean and dry bottle (50ml).

Precaution

The specimen should not be collected in condoms ,  Since the  power or lubricate applied in condoms may be spermici.The container in which the specimen is collected should be free from detergent.

Storage

The specimen should be examin immediately after collection. If it is necessary to dtore then it should be kept at R.T (25 ± 50C )

Sperm are easly damaged by either excessive heat or excessive cold.

 

Morphology of human spermatozoa

Each normal human spermatozoa is made up of two main parts.

1.       A Head :- It is flat and measure measure approximetly 0.5 micrometer in length 3 micrometer in wide and 1.5 micrometer in thick.

 

2.       A Tail  :- The tail measure about 50µm. It consist of 3 parts. The mid piece, The principal piece thr terminal piece.

 

3.       The tail means by which the sperms move.

 

 

 

Precaution

Name of the patient, Date & Time of Sample (semen) collection , Length of absentise and the time of interval between the collection analysis should be reconered in the semen analysis report.

 

Composition

The prostrating fluid in semen is acidic contain acid  phosphate and proteolytic enzymes which acts on the coagulum of the semen vesicles, thus resulting liquefication on the semen problem may their for inferty lesation.

Spermatozoa are cells which fertilized the egg, they are the main focus when performing a semen analysis.

Source of secration                                                                                         % ejaculate

         i.            Testis                                                                                                    5%

       ii.            Seminal                                                                                                46 – 80 %

     iii.            Prostate                                                                                               13 – 33 %

     iv.            Bulbo urethral and urethral gland                                                       2 – 5 %

 

Routine examination of semen

Throughly mix the sample before Examination.

Physical examination :-  Which is performed after a maximum of 60 min by which the time of liquiofication is complete.

1.       Colour and Appearance :- Normal semen is grey white, Vicid and Opque.

Case of Altered semen colour

                                            i.            Yellow Colour – Pyo spermmia (inflammatory cause) billrubin presence.

                                           ii.            Rust Colour (blooding) – Minnar bleeding in the seminal vesical.

2.       Volume :- Normal volume is 2-5 ml per ejaculate.

       Abnormal Volume

                                i.            Hypo spermia - <1.5 ml is called Hypo spermia.

                               ii.            Hyper spermia – Increase in semen volum >5.5 ml (A very rare phenomena )

                             iii.            A Spermia – Means the total Absess ejaculate.

                             iv.            AZoo spermia – Means the Abscess of Spermatozoa in semen.

                               v.            Oligo Spermia – Oligo spermia known as hyper spermia.

 

3.       Viscosity :- Take the specimen in the pasture piyette and expel it. The specimen whith normal viscosity can be poured drop by drop.

Ø  Inccreased viscosity result in poor invasion of cervical mucus as demonstrate by post coital studies.

4.       Liquefication time :- Normally a sample of semen fealiure liquefy in 60 minute indicates in adequate prostratic secresion.

 

 

Chemical Examinattion

Determination of PH :- By Ph paper strip method, normal 7.2 – 7.6 . Cause of ulteration in the ph.

Ø  7< - Obstruction to ejaculatory duct and conjenitial absence of seminal vesical. Such specimen contain mainly prostatic secretions and hence and acitic Ph.

Ø  Lesser then >7 – acute infection of the prostatic (Accute prostaties). Seminal vesicle and epididymis.

Quantitative determination of semen.

This is based on the principal that the fructose react with resorcinol. In the strong acidic medium to give a red colour complete which is compare with the known fructose slandered at 490nm.

Normal range :- 150 – 300 mg/dL

Ø  High fruetose lavel; (> 300 mg/dL) is Associated with low counts.

Ø  Low fruetose lavel (<150 mg/dL) it associated with decreased testrone and respond well to testestrone tharepy.

Study of motility of sperm

a.       Place a small drop of liquefied semen on a glass slide and cover it with cover slide.

b.       Examine the cover slip preparation under the high power objective reduced elimination.

c.       Count the number of active motile sperm and out  of the total count of the sperm.

d.       Culculate the sperm showing the actual progressing motion.

e.       Examine the slide after 2 hrs, 3 hrs and 6 hrs, To present the drying effect of cover slip preparation should be placed under a petridish with moist filter paper.

f.        Observe for pus cells, epithelial; cell , red blood cell and for other finding.

 

Determination of sperm count

a.       After liquification gently mix the specimen.

b.       Draw the semen up to the 0.5 mark of a WBC Pipette.

c.       Draw in the semen diluting fluid to the 11 mark and mix well.

d.       Load the improved neubour chamber and allow the sperm to the settle for about 5 minute.

e.       Count sperm in the four corners sequare.

 

Calculating Sperm Count

The formula for calculating the sperm count, when 5 small squares within the large center square are counted is:

Number of sperm counted x dilution factor/volume x 1000 = sperm/ml.

Example: 50 sperm are counted in the five small squares. The dilution is 1:20.

Sperm/ml = 50 x 20/0.02 mm3 x 1000 mm3/ml = 50,000,000 sperm/ml.

If the dilution is 1:20, and the usual 5 small squares are counted, then the formula can be simplified as follow:

Sperm/ml = Sperm counted x 1,000,000.

 

Composition of semen diluting fluid

 

Sodium bicarbonate                                                                        5 gms

Formalin                                                                                              1 gm

D/W                                                                                                       99 ml

It should be stored at R.T

Normal Range                                                                                   40 – 300 millions/ml

 

Cause of Low count

a.       Infection :- mumps or chetis prostanitis

b.       Obstruction :- Occlusion or absence of efferent duct.

Endo crinopathies :- Hipo pitutarism, hypo gonodism, hypo and hyper throdisim &estrogen secration tumors.

 

Determination of morphology of sperms

a.       After liquification make a thin smear of the semen on a glass (similar to blood smear)

b.       Let it dry in the air and then heat very gently to fix. If necessary remove the mucous by diping the smear in the semen diluting fluid & then in D/W.

c.       Stain the smear by using Leishman stain or stain 0.2% (weight/volum). Aqueous bnasic fuchsin for 5 min.

Normal Observatiom                                                                     Colour

Spermetozoa heat caps                                                                 light blue

Nuclear posterior                                                                             Dark blue

Bodies & Tail                                                                                  Red or Pink

Spermetazoa size                                                                            50 – 70 m

Head size                                                                                         3-6µ x 2-3µ

 

Observation for other abnormalities

         i.            Abnormally shaped head.

       ii.            Abnormally size head (Giant or minut head).

     iii.            Double heads

     iv.            Vacuoles in the chromatin.

       v.            Middle section – Absent, swollen.

     vi.            Tail – May be rudimentary, double or absent.

                                                                     RRANDHIR KUMAR

                                                    RDK PARAMEDICAL STUDENT STUDY

 



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