( ...)The QIAGEN Guide to Animal Cell Culture

Part V: Counting cells

Welcome to the fifth of a series of articles providing useful hints for culturing animal cells. This article contains considerations for counting cells.

This protocol has been adapted from references 13. Protocols in this series are examples of methods for general cell culture and have not been rigorously validated and optimized by QIAGEN. There are many alternative protocols in current use.

Cell counting using a hemocytometer

It is often necessary to count cells, for example, when plating cells for transfection experiments. One method for counting cells is to use a hemocytometer. A hemocytometer contains 2 chambers (Figure 1). Each chamber is ruled into 9 major squares (volume of 0.1 mm3 or 1 x 104 ml each). Cell concentration is determined by counting the number of cells within a defined area of known depth (volume).

1. Clean the surface of the hemocytometer with 70% ethanol or another suitable disinfectant, taking care not to scratch the surface of the central area. Dry with lens paper.

2. Clean the coverslip, wet the edges very slightly, lay it over the grooves and central area of the hemocytometer and gently press down.

Tip - It is important that the coverslip is properly attached to obtain the correct chamber depth. The appearance of Newtons rings (bright and dark rings caused by interference in the air between the coverslip and the glass surface of the hemocytometer) will confirm that the coverslip is attached properly.

3. Harvest the cells, either by trypsinization (adherent cell cultures; see QIAGEN News 2002 No. 3, 20) or
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