organoid imaging

Posted by Mary Ellen Pease on
URL: http://confocal-microscopy-list.275.s1.nabble.com/organoid-imaging-tp7590835.html

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Hi all,

I have a user who would like to image live organoids via confocal/multiphoton without jeopardizing the sample. Has anyone figured out a material in which the organoid could be placed that would eliminate movement/drift during imaging, without damaging it so the sample could go back into culture afterwards? So far, the papers I've looked at on this topic cryoembed them at a certain time point for immunohistochemistry. My user's lab has samples which are expressing GFP and RFP, and would like to look at them in vivo. Someone in our department suggested putting them in a mix of matrigel and culture media, which could slow them down temporarily, but in my experience, matrigel attenuates the signal. We have a multiphoton system on an inverted Zeiss platform (LSM 710). we can image using an objective inverter and dipping lenses, or from the bottom through a coverglass dish. Since we are working remotely at this point in time, I was hoping to get some direction from this group.

Best,

Mary Ellen Pease
Manager, Wilmer Microscopy and Imaging Core Facility (MICF)
Johns Hopkins SOM, Baltimore MD