I'd like to know how to calculate the mean of fluorescence per cell by flow cytometry.
I'm just measuring the total amount of DNA in one wt cyanobacteria spp, and comparing with the total amount of DNA with a modified strain that is supposed to have more DNA (so I should detect more DNA per cell). I'm using SYBR green.
So, I just would like to compare the mean of fluorescence (SYBR) per cell in both strains. I just need relative units (for example, if mutant have double amount of DNA than wt).
In Flow Jo there is a tool called "mean". You can apply this statistic to a particular fluorescence (I understand that this means: this one particular fluorescence/cell, in arbitrary units).
Does anyone know if I can just use that measurement for the publication? Or should I normalize this value somehow? Not sure...
Dear Judith,
if you just want to compare the two groups it should work in the way you described it.
Have you measured both groups with the identical setup of the instrument (e.g. at the same day)? Otherwise I would recommend to include fluorescent beads to ensure the comparability of the measurements.
I would suggest including fluorescent beads as internal standard even if the instrument setups are the same for the two types of cells. Ideally, the standard would be cells with a known genome size, and this standard would be added to the cells you want to quantify. Hoping this will help, Claude
You have MFI buried deep in the add statistics option in flowjo vx u can set your gates and add the MFI
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