Hello everyone,
I am looking to study the fluorescence intensity of GFP in a GRAM positive bacterium. The culture medium is M17+sucrose+glucose. Does this medium fluoresce in the GFP wavelengths? If so, do you know a medium for GRAM positive (lactococcus) that does not fluoresce?
Thank you for your help and support
You can easily test the medium for fluorescence. If it is significat, then wash the cells with PBS before measuring the GFP fuorescence.
Another issue to check on is whether the bacterial cells exhibit fluorescence. They probably do because of such things as flavins and heme proteins, but the question is whether it will be too much to allow detection of GFP fluorescence above the background.
Hi Romain Bernardo , This culture medium is nor fluorescent. Unless you have a contamination in your media or your cells are having auto-fluorescence. As a precaution, you can try with a new media bottle and also make sure your cells are in good condition (less debris or no debris). In addition, also check the microscope settings to see whether you are using a very high power and a gain in your settings which can often produce significant background or autofluorescence artifacts. Hope this helps.
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