07 Januari 2010

Comparing Automated And Manual Cell Counts For Cell Culture Applications

Arun Tholudur, Luis Giron, Kohinoor Alam, Tiffany Thomas,
Eric Garr, Gresham Weatherly, Keith Kulowiec, Melissa Quick, and Scot Shepard


In biomanufacturing based on animal cell culture, cell enumeration and viability determination are often accomplished using the trypan blue dye-exclusion method (1). This method is based on the fact that viable cells exclude the dye and remain visually clear, whereas nonviable cells are stained blue.
Historically, cell counts using the technique have been performed manually using a hemacytometer. Cell culture samples are mixed (after dilution in an isotonic buffer, if necessary) with known volumes of trypan blue dye, then loaded into a hemacytometer and counted under a microscope (usually at 100× magnification). It has long been recognized that cell counts obtained using this procedure are subject to inter- and intraanalyst variability that arises from subjective assessments and cell staining, sample dilution, and

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