I transfected my cells (primary human dermal fibroblasts) with a plasmid containing the gene of interest and a C-terminal turboGFP tag.
I could see the turboGFP fluorescence using a fluorescence microscope.
So I fixed these cells in ice-cold methanol and stored them at 4 °C, covered with an aluminium foil.
However, when I performed an immunofluorescence, the green fluorescence of the turboGFP almost completely disappeared.
Does the GFP fluorescence disappear in fixed cells?
How long does it last?
GFP fluorescence does not work after methanol fixation, however it should work fine after formaldehyde fixation.
Thank you, John Hardy Lockhart
And do you know how long the GFP fluorescence lasts?
Should I perform my experiments soon after formaldehyde fixation? Or can I store my cells at 4 °C without any fluorescence loss (or at least a little loss)?
You should be able to store the cells for quite some time after fixation. Just wrap the plate in aluminum foil to minimize light exposure.
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