Myeloma and Hybridoma Cell Counts and Viability Checks

For most purposes, the number of hybridoma or myeloma cells can be estimated simply by observing the cells under the microscope. When an exact cell count is needed, the number can be determined by using a hemocytometer (improved Neubauer counting chambers are the most commonly used). This is a simple device in which a special coverslip rests on supports that hold it 0.1 mm above the base of the slide. The slide is engraved with a series of lines that form 1 × 1-mm squares. By counting the number of cells within the 0.1-mm³ chamber formed by the 1 × 1-mm square and the height of the coverslip, an accurate quantitation of cells per milliliter can be calculated. To determine the percentage of viable cells within a population, the cell suspension is mixed with a vital dye and observed under the microscope. Vital dyes are excluded from living cells but stain dead cells. The most common dye used for these stains is Trypan Blue.