[OpenSPIM] Heating and CO2

[David Potter]

Matt,

I haven't had a need for CO2 yet so I can't help you there but I have been
successfully heating my chamber to 27C for up to 24 hours. Longer wouldn't
be a problem. Samples are largely zebrafish. The chamber is anodized
aluminum and the liquid volume is 150mL. I cap the chamber to slow
evaporation while leaving a hole for the sample mount to move. I went the
simple route and attached two polyimide encapsulated  copper foils, via
PSA, to the chamber bottom paired with a very low tech controller, an NTC
thermistor and an old 12V laptop power supply. The foils and controller are
from Minco (https://www.minco.com/Components/Shop-by-Product). Your chamber
size will dictate the specific foil but an example would be, say, part
number HK6912. The controller I used is the CT325. Throw in a fuse to keep
OHS moderately happy. You'll need to calculate the thermal mass of your
chamber plus medium (plus anything the chamber touches i.e. transfer heat)
and the warm-up time blah blah and match the parts correctly but that's all
straightforward. My chamber warms up from 18C to 27C in about 10 minutes.
Put the thermistor as close as you can to the sample. I put a temp probe in
an agarose mount and adjusted the CT325 to give me 27 at the probe. It's
not sexy but it works and it was cheap at ~$200 USD.

If I wanted CO2 control, I would probably do nothing fancier than a well
sealed plexi box around the sample chamber and stage. Feed in heated
CO2/air dual gas (or air/N2/CO2 tri-gas) at a slight positive pressure and
you don't need CO2 controllers. I would include an IR sensor (not a wet
sensor) as it's good form and allow record keeping. Add a gas pressure
sensor and wire every sensor into a beagle bone to record all the data. You
might find an old system designed for a microscope nobody wants anymore and
up-cycle the parts. I would put the stage inside the box as a bigger box is
way cheaper and simpler than soft sealing the sample holder arm. Or go
crazy and box everything but the camera and lasers with opaque plexi and
you get gas, temp and safety all in one. Send me photos if you build one.

Good luck.

- David

David Potter
Lattice Light Sheet Microscopy Manager
Monash Micro Imaging - Advanced Optical Microscopy
Monash University Clayton, VIC 3800, Australia



On 30 January 2018 at 05:00, <openspim-request at openspim.org> wrote:

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>    1. Heating/CO2 solutions for sample chamber (PEARSON Matthew)
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> Message: 1
> Date: Mon, 29 Jan 2018 12:26:04 +0000
> From: PEARSON Matthew <matthew.pearson at igmm.ed.ac.uk>
> To: "openspim at openspim.org" <openspim at openspim.org>
> Subject: [OpenSPIM] Heating/CO2 solutions for sample chamber
> Message-ID: <E944BD3E-A8A2-4190-8BBD-B04593DE95D4 at exseed.ed.ac.uk>
> Content-Type: text/plain; charset="utf-8"
>
> Hi all,
>
> I was wondering if anyone has any advice/experience they'd like to share
> on how to implement heating (up to 37C) and 5% CO2 supply to a SPIM sample
> chamber.  What samples  do you image and over what kind of time frame?
>
> Thanks for the help,
>
> Matt
>
> --
> Matt Pearson
> Microscopy Facility
> MRC Human Genetics Unit
> Institute of Genetics and Molecular Medicine (IGMM)
> University of Edinburgh
> Crewe Road
> EH4 2XU
>
>
>
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