Does anyone have any ideas about high refractive index mounting media for either fixed, immunolabeled samples or for living, GFP-fusion expressing cells, to avoid spherical aberrations with high numerical aperture objectives? Media with RI over 1,5 would be best.
you are describing two very different situations.
For fixed immunolabeled samples there are a number of different mounting media (Prolong Gold, Vectashield, Mowiol, etc) with high refractive indices (~1.45-1.5) that approximate the RI of coverslip glass.
For live cell imaging the situation is different as you can't really replace the aqueous medium with mounting medium without killing your cells. How thick is your sample? If you are only imaging cells, the simplest situation is to grow them on glass-bottom dishes (e.g. MatTek). If you are imaging close to the coverslip surface (i.e. within a few microns), you should do fine with a standard oil immersion lens on an inverted microscope as the oil will be close to the coverslip RI. If, however, you are imaging live cells in aqueous medium far from the coverslip, you may do better with a water immersion objective. See http://www.microscopyu.com/articles/optics/waterimmersionobjectives.html for a more detailed discussion of imaging at depth in aqueous solutions.
To add to this, you need a water immersion lens with a correction collar if you are using a coverslip to image live cells with minimal aberrations.
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