What kind of system would you recommend for live-cell imaging? Would you use live-cell incubation chambers or something else. Also what are the advantages of live-cell imaging?
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Fluorescence detection in living cells and ways to improve your image
Company: BioTechnical Institute of Maryland
Job Title: Director
There are lots of good chamber systems for live imaging. If imaging is occurring for a few hours you might be able to get away without an incubation chamber but if your incubations are longer a more sophisticated system would be necessary. Here is a good website from Nikon that talks about them (http://www.microscopyu.com/articles/livecellimaging/culturechambers.html). Obviously a good chamber needs to maintain incubator like conditions without condensation and the viewing port needs to be optically clean. Also a good chamber will have some ports for media perfusion as wells a heating system to maintain the appropriate growth temperature. Typically some sort of glass is used since cell culture plastic does not work well with fluorescence. Live cell imaging allows one to visualize changes in the cells over time and with appropriate tags it is possible to follow trafficing of proteins or complexes as well as visualize interactions. One of the biggest problems that make live cell imaging difficult is the background fluorescence from the cell culture medium itself. Most would opt for media without phenol-red but even this has significant background problems. This is one reason people use DPBS for this type of microscopy. To answer some or most of these problems with auto-fluorescence and poor signal:noise, GIBCO is about to come out with a new product specifically designed to address these problems. A new product called FluoroBrite DMEM is being released that has background equivalent to DPBS and 90% less than standard phenol red-free DMEM. The nice thing about this is you can maintain optimal cell health while getting the images you want with low background.