DIFERENCIAÇÃO E DESENVOLVIMENTO

ANO LETIVO 2022/2023

Sperm Motility Protocol

Motility is divided into 3 categories:

• Progressive motility (MP)
• Non-progressive motility (in situ; NP)
• Immotility

1. Mix the sperm sample well and place a drop of 10 µL on a slide. Cover with a
coverslip;
2. Count at least 100 under the microscope at 100x or 400x magnification once the
sample has stopped drifting (several different fields must be observed);
3. Record the percentage of sperm in each category.

Sperm Morphology Protocol

1. Prepare a smear from a 10 µL sperm sample on a slide and allow it to air dry;
2. Fix the preparation by immersing the slide in a methanol solution (15s);
3. Stain the preparation by dipping the slide in:

- eosin solution (15s)

- thiazin solution (15s);

4. Allow the slide to air dry;
5. Glue the slide to a coverslip using a resin;
6. Count at least 100 sperm cells under the microscope at 400x magnification,
observing several different fields;
7. Record the percentage of normal shapes.