The stained cells are the 'non-viable' cells because the dye has been able to penetrate the cells, ie. In contrast to TB, EB was less toxic to cultured HL-60 cells during the cell counting process. Trypan blue evaluates cell viability through imbalance of the wall and / or cell membrane. EB was found to be an effective dye for accurate counting of cells with different viabilities across three different automated cell counters. It is also toxic to cells causing changes in viability not related to the health of the sample. Consequently, this process is highly subjective. Over the years, there has been an understanding that trypan blue is inaccurate for cell viability under 80 without scientific support. The dye is excluded from membrane-intact live cells, but can enter and concentrate in membrane-compromised dead cells, rendering the cells dark blue. The dye is excluded from membrane-intact live cells but can go through and concentrate in. Trypan blue has long been the gold standard for staining dead cell to determine cell viability. The process is imperfect and cell color is often difficult to interpret. Trypan blue has been the benchmark for staining dead cells to establish cell viability for a long time. We found that cell counting accuracy with EB was comparable to that with TB. Trypan blue is used to count dead cells the dye is excluded from cells with intact membranes so only dead cells are stained blue. SAFETY DATA SHEET Revision Date 1 00654 07050 - Trypan Blue Revision Number 2 1. However, EB is less popular than TB and is seldom used with automated cell counters. Similarly to TB, EB stains only nonviable cells with disintegrated membranes. Erythrosin B (EB) is a vital dye that is impermeable to biological membranes and is used as a food additive. To avoid this chemical hazard, it is of importance to introduce an alternative non-hazardous vital dye that is compatible with automated cell counters. The TC20 cell counter can count single or mixed cell population samples. The cell counter detects the presence of trypan blue in the sample with no user input required and automatically accounts for the 1:1 (trypan blue:cells) dilution factor. However, the wide use of the traditional vital dye trypan blue (TB) raises environmental and health concerns due to its potential teratogenic effects. Only 10 l of cell suspension is required for the cell count. Recent advances in automated cell counters enable us to count cells more easily with consistency.