Total Nucleated Cell and Viable Cell Counts

Accurate cell counts are important when conducting all experiments. Ideally BOTH the total nucleated cell (TNC) count and the viability of the cells should be assessed. Researchers generally use Trypan Blue for both TNC and viability cell counts. However for samples containing red blood cells (RBCs), it is often difficult to distinguish nucleated cells from RBCs using Trypan Blue alone.

For cell samples that contain RBCs (i.e. whole blood, bone marrow), 3% Acetic Acid with Methylene Blue (Catalog #07060) should be used to perform TNC counts. The acetic acid lyses red blood cell and white blood cell membranes, leaving white blood cell nuclei stained lightly with methylene blue. Refer to the Product Information Sheet (available under the Technical Resources section of the product page) for directions for use.

For samples with possible compromised viability (i.e. frozen samples), one should always determine the viability using Trypan Blue (Catalog #07050). In the Trypan Blue Exclusion Test, viable cells with intact cell membranes remain unstained, while the non-viable cells with leaky membranes take up the trypan blue dye. Since Trypan Blue is a cytoplasmic stain, acetic acid-lysed cells cannot be used for Trypan Blue staining. Refer to the Product Information Sheet (available under the Technical Resources section of the product page) for directions for use.