Eosin 0.67g,
Nigrosin 5.0g Distilled water to 100 ml.
The eosin
nigrosin stained slide was used for the estimation of morphological defects under 100x oil emersion lens of light microscope.
Morphological observation was determined from a total count of 100 spermatozoa in smears obtained with
nigrosin and eosin for live-dead count.
Microscopic examination included 10% KOH mount, Gram stain, wet mount & Negative staining by
Nigrosin depending upon the type of sample.
Evaluation of the one step eosin
nigrosin staining technique for human sperm vitality assessment.
Following a 30 second incubation, 150 [micro]L of ten percent
nigrosin solution is added, and mixed.
Viability testing requires a very simple two-step staining procedure, using eosin-Y as the stain and
nigrosin as a counterstain.
The strips were stained with
nigrosin (Acid Black 2), dried, and made transparent with microscope immersion oil that had a refractive index of 1.474.
Determination of dead spermatozoa: For dead sperm ratio, Eosin and
Nigrosin stain was prepared, mixing 1% Eosin and 5%
Nigrosin in 3% sodium citrate dehydrate solution (Zemjanis, 1970).
KG, Karlsruhe, Germany) to four parts 10%
nigrosin aqueous solution [Sigma-Aldrich, USA]) using a glass rod to produce a smear on the slide.
Catalase negative colonies that showed the presence of capsule on
nigrosin stain and were optochin sensitive was further confirmed by bile solubility test.
