Re: organoid imaging

Posted by Mary Ellen Pease on
URL: http://confocal-microscopy-list.275.s1.nabble.com/organoid-imaging-tp7590835p7590854.html

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Thanks very much! I will look into this as well.
take care,

*Mary Ellen*

Mary Ellen Pease
410-292-2551 (cell)
[hidden email]


On Wed, Apr 29, 2020 at 9:53 AM Cammer, Michael <
[hidden email]> wrote:

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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
>
> [hidden email]<mailto:[hidden email]>
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> ________________________________
> From: Confocal Microscopy List <[hidden email]> on
> behalf of Zbigniew Mikulski <[hidden email]>
> Sent: Wednesday, April 29, 2020 2:55:09 AM
> To: [hidden email]
> Subject: Re: organoid imaging
>
> [EXTERNAL]
>
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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
>