Hi everyone,
I am learning flowcytometry and to use flowJo software. I have done my first test trial of flowcytometry from Mesenchymal stem cells. My forward and side scatter values are between
10(4) - 10(6). I do not know whether it is technically correct? If it is wrong how should I correct it. Where can I find troubleshooting information?.
Thanks in advance
Dear Murugadas!
Please you look at these guidelince: (p 1756)
Article Guidelines for the use of flow cytometry and cell sorting in...
Gating strategy for flow cytometry analysis of MSCs. FSC/SSC: forward scatter/sideward scatter. Isotype AB: isotype control antibody. The bar defines the border of 99.5% of the isotype control fluorescence signal. In this example, 99.46% of the MSCs express CD73
Article Fluorescence activated enrichment of CD146+ cells during exp...
Hi Murugadas,
In flow cytometry you obtain relative values, based on the signal intesity (in your case size and optical homogeneity) AND the gain applied to the photodetectors. It also depends on the scale you are using : lin or log.
Cell size and optical granulosity are also dependent on osmotic pressure, while fluorescence can vary with pH and ionic strength.
It is thus impossible to give you range of FSC or SSC values for a cell type.
Hope this helps,
Cheers
Oliv
I would recommend you to have "undyed" control. Measure them at you measurement parameters and adjust forward and side scatter accordingly.
There is no restrictions for you to set FSC and SSC voltage upto your cells/ population is present in the plot.
As nothing in flowcytometry is absolute, it's relative. Relative size, relative granularity, relative expression of any marker.