Fluorescent microscopy uses the emissions of fluorescently labelled probes to generate an image.

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How to prevent e coli from clumping (for FACS)?

I'm performing FACS on e coli, but the cells are clumping together so each event is multiple cells. I ran a control where I had one flask of e coli expressing GFP, and one flask expressing RFP. Run ...
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27 views

background extraction in microscope analysis

I have taken microscope pictures for proteins in HeLa cells, in order to quantify the intensity (immunofluorescence). The proteins are both at the nuclear envelope, and in order to quantify the ...
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57 views

Removal of the Initial Methionine in Venus for FRET

I'm working on building some FRET reporters. In addition to a cleavage site (of varying composition from 15-18AA), a 1 AA linker, I'm using Venus and Cerulean. Initially I was worried that 18AA ...
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Red transcriptional reporter in Psudomonas putida

I need to build a transcriptional reporter in P. putida with high excitation wavelength,in order to decrease phototoxicity (I have been using mNeonGreen until now, and I would like to try something ...
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What does bone look like while fluorescently staining nerves

I am working on a project involving changes in innervation in the subchondrial bone. Nerves were stained fluorescently for PGP9.5 and I have to analyze the samples. However, I don't have much of a ...
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40 views

Effects of Fixation on Cell Membrane

Why does fixing of cells with something like formaldehyde disrupt the membrane enough for probes, antibodies, and dyes to get into the cell?