Posted by
Michael Doube-2 on
URL: http://confocal-microscopy-list.275.s1.nabble.com/organoid-imaging-tp7590835p7590839.html
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Hi Mary Ellen,
Some time ago I was helping a user with their zebrafish embryos. The system was an LSM 780 upright with dipping lenses. The embryos were maintained in position by pressing small dimples into the top surface of (I think) agarose, then resting each embryo in a dimple, covered in water. It was sufficient in most cases to prevent drift.
Best regards,
Michael
On 28/04/2020 22:21, Mary Ellen Pease wrote:
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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
--
[Jockey Club College of Veterinary Medicine and Life Sciences - City University of Hong Kong]
Dr. Michael Doube
BVSc, BPhil, PhD, PGCert Vet Ed, MRCVS, FRMS
Associate Professor
Department of Infectious Diseases
and Public Health
5/F, Block 2, To Yuen Building
31 To Yuen Street
Kowloon, Hong Kong
Tel: (852) 3442 5296
Fax: (852) 3442 0589
Website: www.cityu.edu.hk/ph<
http://www.cityu.edu.hk/ph>
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