Calculating sperm concentrations











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Determination of sperm concentration for small

The semen must be killed to prevent movement and diluted before loading into the hemacytometer. If you have used a phase type hemocytometer the thin ones then be sure the phase rings are in and the condensor is all the way up. The objective is to kill sperm and to dilute them sufficiently so that you will count - sperm cells in the 5 squares as described below. This number is one of the most predictive of eventual pregnancy, according to several studies. The number, in millions, of spermatozoa per milliliter of semen. The tip of the pipette is placed in the V-shaped groove on the hemacytometer to load the sample into the chamber about 15 microliters. When counting, count only those cells on the lines of two sides of the large square to avoid counting cells twice. This surface has a grid etched into it. The hemocytometer is 0. The example at right shows red lines where cells on the line would be counted. You may also need to raise or lower the condensor so you can see the sperm. The chambers are overlaid with a glass coverslip that rests on pillars exactly 0. Multiply the mean obtained in 1 by 10, to obtain the number of cells per ml of diluted sample.

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The formula for calculating the sperm count, when 5 small squares within the large center square are counted is: Number of sperm counted x dilution.

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Description: Each of the nine squares on the grid, including the central counting area of 25 large squares, has an area of 1 square mm, and the coverglass rests 0. Complete no masses or strands. This helps to hold down the coverslip while loading the sperm. Therefore you must multiply your count in 5 squares by 50, in order to determine how many sperm would have been in 1. Count sperm heads in the 5 large squares as indicated by the green color. The dilution factor must be recorded to allow calculating the concentration. Loading the Hemacytometer. Dilute sperm with water. Male Fertility Lab University of Washington. It is important not to overload the chamber, as doing so will give an inaccurate count. The rails are designed to hold a coverslip 0. All 25 large squares can be counted, or a counting pattern using fewer squares can be used like the ones below.
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