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DisGUVery / disguvery

GNU General Public License v3.0
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Issues with encapsulation efficiency #4

Open m-sree opened 8 months ago

m-sree commented 8 months ago

Hi,

I've been using DisGUVery's encapsulation module for analyzing negative control GUVs, where no encapsulation is expected. However, I've encountered an issue where the analysis yields negative values for encapsulation efficiency. Could you provide guidance on adjusting the software or analysis parameters to address this? Specifically, I'm interested in understanding the best approach for background correction in this context, and whether there's a recommended method for handling or interpreting these negative values.

Thank you for your assistance.

Best regards, Sree

cristina-mt commented 8 months ago

Hi Sree, When you are talking about no encapsulation, does this mean that all the dye is outside the vesicles, but it's still visible in the image? i.e. the fluorescence signal is visibly higher outside? If this is the case, you would see negative values with any type of background correction. This just shows that you have less dye inside the vesicles than the one you have outside (which makes sense if you have no encapsulation)

However, if the fluorescence signal outside the vesicles is lower than inside, then we'd need to run some tests with your type of data and analysis flow to check if there's a bug.