When I perform cell cycle analysis using Cell Quest flow cytometer, I can not well initial gating of FSC-H and SSC-H. Because most of the cells are not see in the FSC and SSC figures. If anyone have experience of initial gating in cell cycle using PI, please help me.
Bertrand Cornu
Check your sample preparation and staining protocols: Make sure that your samples are prepared and stained correctly, as this can affect the distribution of cells in the FSC-H and SSC-H plots. For example, if your cells are clumped or aggregated, this can cause the FSC-H and SSC-H distributions to appear distorted or shifted. Adjust the scale of the plots: Try adjusting the scale of the FSC-H and SSC-H plots to see if this helps to improve the separation of the cell populations. You can also try using a logarithmic scale to see if this helps to better distinguish between the different cell populations. Use alternative markers: If you are having difficulty gating cells using FSC-H and SSC-H, you may want to consider using alternative markers to help distinguish between different cell populations. For example, you could try using DNA content or cell cycle-specific proteins (e.g. cyclin B) to help distinguish between different phases of the cell cycle. Use manual gating: If all else fails, you may need to resort to manually gating the cells using the FSC-H and SSC-H plots. This can be time-consuming, but it allows you to carefully inspect the distributions of cells and manually select the populations that you are interested in.
Gating cells in flow cytometry can sometimes be challenging, especially when the distribution of cells is not well separated in the FSC-H and SSC-H plots. There are a few strategies that you can try to improve the gating of cells in your cell cycle analysis:
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Aung phyo Wai
Thank you so much. I am using FACScalibur (cell quest) software and this is first for me to detect cell cycle analysis. I did not how to proper adjusting the initial gating of cell population.
I will try again.
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