We have developed two SFT Kits for assessment of Sperm Morphology - PAP Staining Method and Diff-quick Staining Method.
We have introduced Spray bottle for reagents for PAP Staining Method which gives immense ease of procedure. Also our Diff-quick staining kit takes very short staining time and gives good contrast.
Accurate evaluation of sperm morphology is possible, only when these basic steps are followed.
(1) Appropriate preparation of slides & smears
(2) Good fixation
(3) Proper staining of smears
(4) Using suitable magnification
(5) Evaluating recommended numbers of microscopic fields and sperms
(a) The glass slide used must be thoroughly clean & dry. In order to obtain thin and even spread sperm smear use recommended ‘Semen Drop’ volume depending on sperm concentration. Use 10-15 µl droplet if the sperm concentration is > 20 million/ml; with concentrations < 20 million/ml, a 5-10µl droplet is recommended.
It is a preferred practice now to prewash the semen sample & adjust sperm concentration. Prewashing of semen sample helps to remove excess seminal plasma and improve sperm concentration.
(b) The fixation procedure followed is dependent on the staining method employed. In general, prepare thin semen smear, let it air dry & then fix it in a fixative, which is again air dried.
(c) Ideally, modified PAP staining is preferred. However, Diff-Quik staining is also used because of its simplicity, short staining time & good contrast.
(d) Magnification of 100x objective (Bright field) lens is preferred for evaluation of stained semen smear.
(e) With modified PAP staining, acrosome stains as light-blue and the post-acrosomal region of head as dark blue; while mid-piece stains green and tail as red.
(f) In Diff-quik staining, acrosome stains as pale purple, while post acrosomal part of head, mid-piece & tail stain as dark purple. Diff-quik staining results into slightly swollen heads.
(g) It is recommended to evaluate a minimum of 100 sperms from different microscopic fields.
(h) Normal sperm morphology has been shown to be predictive of ‘Male Fertility’ independent of the other semen parameters. Sperms slightly abnormal or borderline forms must be considered as abnormal.
|Semen Analysis Chamber (Sperm Meter)||Semen Collection Container|
|Controlled Temperature 370C Dry bath (Sperm Warmer / Water bath)||Test-tubes|
|Centrifuge Machine (Androspin)||Glass-slides|
|Slide-warmer||Non-spermicidal Semen Collection Pouch (Sperm Collect)|
|Glass Slide Stand||Coverslips|
|Stop-watch (Andro Watch)||Pasture Pipettes|
|Set of Pipettes||Filter Papers|