Rh Factor: Rh-Negative is on the left, Rh-positive is on the right.

Hemacytometer

How to count with a hemacytometer

Allow the cell suspension to settle for at least 10 seconds.

Count all of the cells in each of the four 1 mm³ corner squares labeled A thru D in Figure 1 on the next page.

DO count the cells touching the top or left borders.

DO NOT count the cells touching the bottom or right borders.

Determine the Cell Count.

Calculate the total cells counted in the four corner squares.

If the total cell count is less than 100, or if more than 10% of the cells counted appear to be clustered, carefully re-mix the original cell suspension and repeat steps 2 through 4 (above).

If the total cell count is greater than 400, dilute the suspension so the count will be 100-400 cells. Then repeat Steps 2 -4 (above).

Calculate the cell count using the equation: cells/ml = (n) x 10⁴,

where: n = the average cell count per square of the four corner squares counted.

Example:If the calculated average (n) of cells in the four 1 mm corner squares of the hemacytometer is 30:

cells/ml = (n) x 10⁴ (or) cells/ml = 30 x 10,000 = 300,000 cells/ml.

Determine the total number of cells in the total suspension volume.

Determine the total volume of the cell suspension.

Multiply the volume of the cell suspension by the "cells/ ml" value calculated above.

Example:If the initial suspension volume is 2 ml:

cells/ml x total volume = 300,000 cells/ml x 2 ml = 600,000 cells.