Download: Viability Protocols for Blood-based Samples
Impact of RBC contamination in clinical samples
- Many blood-based samples (whole blood, peripheral blood, bone marrow, PBMC, cord blood, etc…) may contain residual red blood cells even after RBC lysis. (See figure below)
- When samples are enumerated using manual counting, the presence of residual RBCs can lead to inaccurate cell concentration and viability readings.
- Nucleic acid dyes, acridine orange (AO) and propidium iodide (PI), only stains nucleated cells and therefore yields accurate and consistent results.
- The Cellometer image cytometers acquire multiple images and automatically reports the concentration of nucleated cells and percent viability.
- Live, AO positive, white blood cells (WBCs) are green
- Dead, PI positive, WBC are red
- In this PBMC sample, red circles indicate contaminating RBCs that are seen in the bright-field image but are not stained with AO/PI.
- The Cellometer accurately and automatically reports the cell concentration and viability in less than 30 seconds!