Hi-Tech Solutions

Sperm Morphology Test Diff – Quik Method(1) (2)Intended use and principle The Diff-Quik Staining kit contains rapid stain solutions for Semen smear. Stained in only 15seconds. By varying the number of dips in the appropriate staining Solution, different degrees of shading and intensity are easily obtained.Reagents Buffer solution Fixative Solution Stain Solution IStain Solution II Warning*Fixative Solution contains Methanol >50%*Toxic by inhalation and if swallowed.*Keep container tightly closed.*Keep away from sources of ignition – No smoking.*Avoid contact with skin and eyes.*In case of accident or if you feel unwell, seek medical advice immediately.*Stain Solution I contains sodium azide, it may react with lead and copper plumbing to form highly explosive metal azides. If discarded into sink, flush with a large amount of water toprevent azide build-up.Stability If stored appropriately at 15-30ºC all 4components of the staining set are stable after the first opening until the indicated expiration date. The stability is monitored throughout the dating period. Procedure Take 1 ml of liquefied semenAdd 1 ml of Buffer Solution, mix well ¢rifuge at 2000 RPM for 2 - 3 minutesDiscard supernatant (Seminal Plasma) andadjust the sperm concentration between 40 - 60millions / ml with Buffer solution & use 5 μLto prepare the smear. If adjusted concentrationis less than 40 millions/ml, use 10 μL toprepare the smear. Allow smear to air dry.Put and cover entire smear with 1 ml ofFixative Solution. Keep it for 5 seconds.Drain off the fixative solution and allow smearto air dryPut and cover entire smear with 1 ml of StainSolution I. keep it for 5 second.Drain off stain solution I.Put and cover entire smear with 1 ml of StainSolution II. Keep it for 5 second.Drain off stain solution II.Rinse slide with distilled water.(3)Drain off distilled water and clean the back sideof the slide with the Filter paper.Allow slide to air dry and proceed withdifferentiation.ExaminationPut a drop of immersion oil on dry smearExamine the smear under microscope with thehelp of 100x lens. Examine 200 sperm andevaluate followings as per WHO criteria:- Normal Sperm- Abnormal SpermIn abnormal sperm, detailed abnormality(defects) pertaining to head, mid piece and tailshould be noted down.RemarkThe intensity of the staining can be varied byincreasing or decreasing the volume of theStaining solution and timing, but never gobelow 5 seconds.(4)To increase red staining, increase the Volumeof stain or time in Stain Solution I. To increaseblue staining, increase the Volume of stainingor timing in Stain solution II.If the standard procedure results in a toointensive Red or Blue staining, the pH of thedistilled water should be checked.Diff-Quik Solutions should be changed ifstaining differs from normal.WarrantyThis product is warranted to perform asdescribed in its labeling and in the productliterature and HI-TECH SOLUTIONSdisclaims any implied warranty ofmerchantability or fitness for any otherpurpose, and in no event shall HI-TECHSOLUTIONS be liable for any consequential damages arising out of aforesaid expresswarranty.

Acrosome Intactness Status
Take 500 Micro Litre Of Hts – Acrosome Solution In A Test Tube, Add 100 Micro Leter Liquified Semen Sample To The Centrifuge Tube And Mix Well.incubate At 37degree For 30 Minutes. Put A Drop Of Mixture Solution On The Gcar Slide & Make Smear Very Smoothly, Air Dry For 2 Minutes To Remove Excess Moisture.
Incubate For 2 Hours At 37 Degree Centigrates With Moisture Chamber.
Air Dry And Examine Under 10x – 40x Objective.count The Spermatozoa With Halos.
Normal Percentage > 50%

Sperm Vitality Test 
Procedure:
1 drop of liquified semen 
2 drop of Esoin stain and mix well
3 drop of Nigrosin stain mix well
put one drop on plain slide and prepare smear
air dry and see under 100 x objective with oil emmersion  

Result: white sperm alive, Red sperm dead.