A review of cell counting methods pre- and post- levitation.
Obtaining accurate cell counts is critical prior to and following viable cell enrichment and debris removal via levitation. Cell counts may be obtained via hemocytometer or automated cell counter; LevitasBio recommends Nexcelom in conjunction with live dead viability stain such as AO/PI for determining cell counts. Other cell counters would be perfectly suitable, however it is critical to understand the operation and limitations of each automated cell counter. It is recommended that users review the user manual for their particular cell counter to ensure they understand the counting range, input requirements, and limitations of the automated cell counter, minimum and maximum number of cells that fall in the linear counting range. Another aspect to consider is how well the automated cell counter works with debris-laden samples and other challenging samples types such as single cell suspensions derived-from tissue. Finally, output samples from LeviCell are typically concentrated in <100ul, so consider if you counting method will require a large volume of your output sample. If using a hemocytometer, see the counting tips and guidelines below. Whether using manual or automated counting methods, when in doubt count your samples in duplicate or triplicate.