Re: organoid imaging

Posted by Mary Ellen Pease on
URL: http://confocal-microscopy-list.275.s1.nabble.com/organoid-imaging-tp7590835p7590853.html

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thanks George - i was hoping an on campus expert would be listening! I will
look into the manuscript, etc. I'm home for how and I don't know if/when we
will be ready to try again since everyone on campus is in shutdown - i know
the 2 labs I was working with have put things to be for now. These are
interesting times we live in.

my jhu address is [hidden email].

Best,
*Mary Ellen*


On Tue, Apr 28, 2020 at 11:13 AM George McNamara <[hidden email]>
wrote:

> Hi Mary Allen,
>
> Jawad Allen -- now Dr. Jawad Allen -- in Cindy Sears lab did long term
> (weekend) imaging on our Ross Bldg (9th floor)  Olympus FV3000RS
> confocal inverted (IX83) microscope (OkoLab stage top + plexiglas shroud
> incubator, 37 C, tage top humidifier & optional 5% CO2 ... 10x/0.40 WD
> 3.1 mm or 20x/0.75 WD 0.6 mm, if I recall correctly ... standard lenses,
> so equivalent on your LSM710MP would work fine),
>
> http://confocal.jhu.edu/current-equipment/fv3000/
>
> (I believe) embedded in matrigel, similar to Philipp Tripal's reply. You
> should be able to get full details from Cindy (i.e. manuscript in prep).
>
> Yes, Jawad brought his SBS plates between buildings (in an appropriate
> container). I strongly recommend the research lab have a quarantine
> incubator for plates that leave the lab and return.
>
> Your user is welcome to try out our FV3000RS for comparison - once JHU
> SOM re-opens. Full set of laser lines from 405-730 nm. We'll probably be
> busy all summer catching up, but should be able to squeeze your user in.
>
> I encourage use of thin bottom SBS plates or 35 mm imaging dish. Mattek
> (and some others) offer #0 coverglass bottom, which provides an extra
> ~70 um working distance compared to #1.5 coverglass (nominally 170 um;
> ibidi's bottoms are typically their image quality plastic, ~180 um).
>
> George
>
> p.s. my JHU email handle is gmcnama2
>
>
> On 4/28/2020 10:21 AM, Mary Ellen Pease wrote:
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > Post images on http://www.imgur.com and include the link in your
> posting.
> > *****
> >
> > 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
>