Have you ever started a flow cytometry run and you cannot see any cells? This can be an alarming feeling, but some straightforward troubleshooting can usually help you find your missing cells. Check out these three questions to tackle common issues associated with disappearing cells.
1.How clean is your machine?
Flow cytometers must be properly maintained in order to assure that the fluidics system is functioning properly. The sample injection port can get clogged, and the sheath fluid may need to be refilled. If you can’t see your cells, running through cleaning and set up procedures can typically resolve common fluidics problems.
2. How are the lasers?
The optics components within a flow cytometer are essential to the function of this machine. Laser alignments can shift or issues may arise with the equipment that detects fluorescent emissions (filters, mirrors, and lenses). It is critical to run through set up procedures, including the use of calibration and compensation controls, to assure that the flow cytometry instrument settings are correct and will detect your samples.
3. Do you have good gates?Cells of interest can be identified and measured on a flow cytometer by using a good gating strategy. During the set up phase of a flow cytometry run, users will have a set of control cells that are used to establish a series of gates around cells with the correct phenotypes. An appropriate gating strategy also assures that dead cells are not included in cell counts targeted during the acquisition phase.
Hopefully, by looking at these three areas, you will be able to find your missing cells. But, if cleaning and maintenance procedures or changes in the instrument settings do not resolve this problem, a flow cytometer technician may need to be contacted. Here’s to finding your cells and having a relaxing summer vacation!