Live Dead Analysis

Hi Hannah,

How do we perform live dead analysis using Biofilm Q?
do we compare the mean intensity of the two channels and will that give us the live dead analysis of the biofilms??


Hi Srinivas,

depending on your data, you can choose several approaches to analyze dead-live dynamics.

I am assuming that you have two fluorescence channels - are these channels exclusive in the sense that no live cells are visible in the “dead” channel and no dead cells visible in the “live” channel? In this case I would recommend to perform a segmentation on both channels, merge them, and look at the dynamics of the properties measuring relative abundance.

If there is one channel in which all cells, live or dead, are fluorescent, I would instead measure the “3D overlap” property after performing segmentation of both channels. This property can be calculated using the fluorescence properties in the parameter calculation tab of BiofilmQ.

It is also possible to measure and compare fluorescence intensities, but I would recommend to only do this if segmentation is not possible for both channels (e.g. if the signal is too noisy in one of the channels). This can also be done via the fluorescence properties (depending on your constellation choose either intensity ratio or mean intensity per cube).

Let me know if you would like to have a more detailed explanation of any the above mentioned approaches - I just wanted to give you a quick overview and description of the possibilities that I am aware of right now.