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Cammer, Michael-2 on
URL: http://confocal-microscopy-list.275.s1.nabble.com/organoid-imaging-tp7590835p7590847.html
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We kept organoids alive and apparently happy for 12-18 hours on a microscope stage using embedding in Matrigel. In addition to organoid imaging alone, cells can be added to Matrigel and watch how they interact with organoids. Imaging in paper below was at low mag, but (not used in the paper below) I fixed and labeled the organoids after overnight imaging to see the same ones at high resolution with confocal, so confident this would have worked live at high res too.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5716041/Small intestinal crypts were isolated and cultured as described previously (Sato et al., 2009<
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5716041/#bib47>). In brief, crypts of proximal small intestine were counted and embedded in 30 µl of Matrigel (Corning) at 10,000 crypts/ml and cultured in DMEM/F-12 ...
Michael Cammer, Sr Research Scientist, DART Microscopy Laboratory
NYU Langone Health, 540 First Avenue, SK2 Microscopy Suite, New York, NY 10016
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Hello Mary,
Have a look at CyGel (you can get it from Biostatus, Abcam also distribute it in the US). It's a liquid at 4C and it gels around 22-27C. It is reversible once you lower the temperature. They claim it's non-toxic. We have not tried with 2P imaging or organoids but it worked quite well in our hands with cells and fluorescence microscopy.
All the best, Zbigniew
Zbigniew Mikulski, PhD
Director, Microscopy Core Facility, Instructor | La Jolla Institute for Immunology | 9420 Athena Circle, La Jolla, CA 92037