ABSTRACT

Fluorescence microscopy is an art, and it will take some practice to be able to take nice images of cells expressing fluorescent proteins. It is always a good idea to take a bright-field image for comparison with the fluorescence image. It is also a good idea to start by looking at the control plate: If it is highly fluorescent under any of the filters, this is background autofluorescence that must be eliminated. Using a neutral density (ND) filter can reduce excitation power enough to cut down autofluorescence without compromising fluorescent protein excitation. A narrower bandpass filter might also be needed; compare different filters as available.