Posted by
jerie on
URL: http://confocal-microscopy-list.275.s1.nabble.com/CO2-grade-for-on-scope-incubator-tp7582397p7582414.html
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Dear Peter,
EMBL operates such incubators for 10+ years using technical grade C02
without any problems. We have even saturated the air in the incubator with
water in order to counteract evaporation of culture media for long time
lapse acquisitions. You can use pre-mixed gas, as mentioned, and it is
better to mix CO2 with air or with N2. A low partial pressure of O2 helps
avoiding photobleaching and facilitates photo-switching of some GFPs.
Kind regards, Jens
Visiting Scientist @ Center for Technological Development in Health (CDTS),
Oswaldo Cruz Foundation (Fiocruz), Ministry of Health, Rio de Janeiro,
Brazil.
http://br.linkedin.com/pub/jens-rietdorf/6/4a3/189/Skype jens.rietdorf
On Tue, Jul 22, 2014 at 8:42 PM, Peter Werner <
[hidden email]> wrote:
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>
> At our facility, we have a spinning-disk confocal system, with an
> incubator that surrounds the entire stage and substage area. The question I
> have concerns the type of CO2 I should be using for the incubator. I was
> told by someone from the UC Berkeley Biological Imaging Facility that I
> should be careful about the grade of carbon dioxide that I use -
> apparently, it should be high purity and as low as possible in H2O. The
> reason is, even if the cell cultures themselves will be fine with a lower
> purity of CO2, CO2 forms carbonic acid in reaction with water, and the
> acidified water content in the chamber atmosphere can be damaging to the
> instrument over the long term, hence, the less H2O mixed in with the CO2,
> the better. If this is the case, that would imply that "Instrument Grade"
> or "Bone Dry" grades would be what I need to use.
>
> I was wondering if anybody else knew of this recommendation and followed
> this practice. It makes sense to me why one would want to minimize carbonic
> acid formation in environments the confocal system is exposed to. On the
> other hand, it does entail some expense, not so much in terms of the gas
> itself, but in terms of special cylinders for high-purity CO2, which must
> be rented, or purchased at $500+ per gas cylinder.
>
> Let me know,
> Peter G. Werner
> Instructional Assistant/Lab Technician, Microscopy, Merritt College
> SEM/AFM Lab Technician, Ohlone College
>
[hidden email]
>