imaging spheroids

Posted by Cromey, Douglas W - (dcromey) on
URL: http://confocal-microscopy-list.275.s1.nabble.com/imaging-spheroids-tp7581903.html

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I am working with a lab that is interested in doing fluorescence microscopy on spheroid assays (clumps of cancer cells seeded and growing inside a moderately thick collagen matrix).  They are looking at a number of different microscopy techniques on campus.  Because our Leica confocal was mostly configured for 2D cultured cell and tissue sections, we have quickly discovered the working distance limitations of our existing objectives.  Our local Leica technical representative will be loaning us Leica's fabulous 25x/0.95 water immersion objective (2.5mm WD) to try out.  This should help with WD issues, as well as spherical aberration in the sample (their dishes do at least have #1.5 thickness glass coverslip bottoms).

Has anyone else worked with these types of assays before?  Any suggestions on the sample prep side or the imaging side to end up with better image data?

Thanks,
Doug

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Douglas W. Cromey, M.S. - Associate Scientific Investigator
Dept. of Cellular & Molecular Medicine, University of Arizona
1501 N. Campbell Ave, Tucson, AZ  85724-5044 USA

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