Immediately transfer the cell suspension to the counting chamber by placing the tip of the pipet at the edge of the chamber and allowing the chamber to fill completely via capillary action. Take a well mixed 20-50 µl aliquot of the dissociated cell suspension using either a Pasteur pipet or a micropipettor only drawing the cells into the tip.Wet the sides of the coverslip with reagent grade water and align the coverslip over the counting chamber.Be careful not to scratch these surfaces. Carefully clean the counting chamber surface and the coverslip of the hemocytometer with 70% isopropanol and allow to air dry.The use of a hemocytometer for cell yield quantitation is outlined however, newcomers to this procedure can refer to more detailed discussions (see Freshney, Culture of Animal Cells, page 227). The use of a cell counting chamber (hemocytometer) for yield quantitation and the use of trypan blue for viability quantitation are recommended. It is important to quantitate the results of each dissociation step in order to effectively evaluate each procedure.
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