|Cell Depth||0.02mm (1/50 mm)|
|Ruling Pattern||Improved Neubauer, 1/400 Square mm|
In order to count the number of sperm per milliliter, dilution is not necessary. To immobilize sperm, there are two recommendations:
Rulings cover 9 square millimeters. Boundary lines of the Neubauer ruling are the center lines of the groups of three. (These are indicated in the illustration below.) The central square millimeter is ruled into 25 groups of 16 small squares, each group separated by triple lines, the middle one of which is the boundary. The ruled surface is 0.02 mm below the cover glass, so that the volume over a square millimeter is 0.02 cubic mm. Count all cells within this center square millimeter (1mm x 1mm area).
The number of sperm per milliliter (ml) = Number of cells counted X dilution (if used) X 50,000*
*50,000 = 50 (cell depth is 1/50) X 1000 (1000 cubic mm = 1 milliliter)
After completing the count, remove the cover glass and clean the counting chamber with water or a mild cleaning solution (10% solution of bleach). Dry the counting chamber with a soft cloth or wipe, or rinse with acetone.