BotuVital is a diluent for sperm evaluation based on Eosin and Nigrosin, which allows the analysis of the sperm pathologies as well as the visualization of plasma membrane integrity in light microscopy at 100x immersion objective.
HOW TO USE:
- Over one of the slide ends, deposit 1 drop of fresh semen (free of diluent) and 1 drop of BotuVital;
- Homogenize the drops until getting a mixture of uniform coloring;
- Smear with help of another slide, slightly loosening the uniform mixture in order to obtain a very thick and dark smear and do not cause mechanical injuries to the sperm, according to the scheme below:
- Wait 5 minutes for the mixture to dry;
- Read in light microscopy at 100x magnification (immersion);
- Count 200 sperm to measure the spermatic pathologies together with stained and unstained ones:
– Sperms stained in pink show damaged plasma membrane.
– Sperms with absence of staining show full plasma membrane.