Processing and staining cells for flow cytometry or cell sorting is a multi-stepped process that can stress cells and cause cell death. Dead or apoptotic cells are troublemakers in cytometry because they can bind antibodies, effectively changing the staining antibody concentration even after titration, which results in suboptimal staining of live cells, and they can autofluoresce, particularly in the green channels, providing false positive signals. Dead cells also do not divide, cannot produce cytokines upon in vitro stimulation, and are often sticky due to extruded nucleic acids resulting is both poor yield and poor purity when sorted. So if you have a large fraction of dead cells in your samples, your results may be significantly skewed or unusable.Read the full article »
Flow cytometry is a valuable research tool because it can generate abundant amounts of very detailed data, but this often means that scientists need to deal with moving and storing large amounts of digital data. In addition, many flow cytometry users run their experiments at a site separate from their lab or work with an offsite contract research organization (CRO) that runs their flow cytometry experiments. This means that flow cytometry data transfer is an issue that researchers must address even in the planning stages of their flow cytometry experiments. Consider these factors related to transferring large datasets for flow cytometry.Read the full article »
We walked you through the basics of achieving your optimal panel design in the “Know your Flow” series, but you probably have more questions as you refine your panel or stumble upon some of the technical nuances of staining cells.
Check out these tips for a successful flow cytometry run.Read the full article »