I have a problem with the scattering gate of the hek 293 suspended cells in flowcytometry, shall I use the log or linear option? what the range to expect my cells appeared in? Do the hek cells have autofluorescence characteristic.
I am here with attaching the scattering dot plot of my cells
Hi Haitham,
In my experience with FACSCalibur flow cytometry - I know you are very likely using a newer cytometer - I believe you should always try the linear scale first. This is because when you use log scale you can "see" everything that is passing through the laser and many times is huge load of garbage!!! Your cytometer must be very clean when you use log scale. I am currently using it with very variable sized cells, such as snake leukocytes. In your case, since you are using a very well stablished cell line, the cell size and scatter are homogenous, linear scale should be better.
Good luck!
Thanks Nicolle, I did it, but cell population percentage is very low; about 28 % of the total applied sample.
Hello Haitham,
I also have two distinct "populations" when analyzing HEK293 cells in flow cytometry. On your figure "Slide1.JPG", you surrounded one population in blue and another one in red. Do you have an idea why we have such distinct populations ?
Thanks in advance for your answer!
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