Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
Date: Wed, 26 Feb 2003 10:10:38 +1100
Reply-To: Confocal Microscopy List <[log in to unmask]>
Sender: Confocal Microscopy List <[log in to unmask]>
From: Stephen Cody <[log in to unmask]>
Subject: Re: live cell imaging
Content-Type: text/plain; charset="US-ASCII"
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
Dear Judy,
I use a premixed 5% CO2 /air mixture, the gas company makes up to order.
They also sell a stock 5%CO2 / oxygen. As my incubator recirculates, I
don't want 95% O2 in contact with the heater element for fire safety
reasons.
It is not necessary to fill the whole chamber with your CO2 mixture.
Just place a Petri-dish lid over your specimen and have a fine gas line
(I use 0.96mm PE tubing) very gently bubbling under the Petri-dish.
Or better still grab a large Petri-dish, cut a hole in the bottom to
accommodate your cell bath. Build up a water-proof "dam wall" around the
hole with silicone sealant, to make a moat. Fill the moat with water and
cotton wool. This will keep the small chamber at saturated humidity.
Bubble your gas mixture very slowly into the water in the moat.
_________________________________
| |
|| ||
|_________|| ||__________|
\
\
Silicon dam
Stephen H. Cody,
Microscopy Manager,
Central Resource for Advanced Microscopy,
Ludwig Institute for Cancer Research,
Post Office Royal Melbourne Hospital,
Parkville, Victoria 3050, Australia.
Tel: +61 3 9341 3155 (BH)
+61 3 9341 3158 (@work AH)
Fax: +61 3 9341 3104
email: [log in to unmask]
http://www.ludwig.edu.au/labs/confocal.html
> -----Original Message-----
> From: Judy Trogadis [mailto:[log in to unmask]]
> Sent: Wednesday, 26 February 2003 6:48 AM
> To: [log in to unmask]
> Subject: live cell imaging
>
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Hello Confocalists:
>
> I have a question for those doing live fluorescent cell imaging in a
> controlled environment over a period of many hours:
>
> We have a plexiglass incubator installed around the stage of a Nikon
> inverted microscope, setup for live cell imaging and are looking for a
> device to both regulate and measure the amount of CO2 inside the
chamber.
> Does anyone know of such a product? Phenol red in culture media can
> interfere with optical quality, yet we have to maintain a stable pH in
the
> solution. I prefer not to have a probe directly in the dish because
often
> the dish will be closed, therefore, measuring the atmosphere is more
> practical.
>
> Also, thinking about which gas to use, I thought of a pure CO2 tank
but
> someone said it may make the environment hypoxic if the exhaust is too
> close to the dish. If the end of the tube (CO2 is bubbled through
water)
> is too distant from the dish, the gas can escape through numerous gaps
in
> the setup and we'll be going through tanks on a daily basis. Would a
5%
> CO2/air mixture be better?
>
> Thanks for any help
> Judy
>
> Judy Trogadis
> Bio-Imaging Coordinator
> St. Michael's Hospital, 8Queen
> 30 Bond St.
> Toronto, ON M5B 1W8
> Canada
> ph: 416-864-6060 x6337
> pager: 416-685-9219
> fax: 416-864-6043
> [log in to unmask]
Stephen H. Cody
Microscopy Manager
Central Resource for Advanced Microscopy
Ludwig Institute For Cancer Research
PO Box 2008 Royal Melbourne Hospital
Parkville Victoria 3050
Australia
Tel: 61 3 9341 3155 Fax: 61 3 9341 3104
email: [log in to unmask]
www.ludwig.edu.au/labs/confocal.html
www.ludwig.edu.au/confocal
-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On
Behalf Of Judy Trogadis
Sent: Saturday, 29 October 2005 6:00 AM
To: [log in to unmask]
Subject: heating live cells
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
To the wonderful Confocal Listserv:
Thank you for the many useful replies with suggestions on regulating
the temperature in our incubator. Of course, off-the-shelf products are
great, however, it was astonishing yet reassuring to know that
individuals are still willing to spend the time to create novel ways of
achieving the same results.
Gratefully yours,
Judy
Judy Trogadis
Bio-Imaging Coordinator
St. Michael's Hospital, 7Queen
30 Bond St.
Toronto, ON M5B 1W8
Canada
ph: 416-864-6060 x6337
pager: 416-685-9219
fax: 416-864-6043
[log in to unmask]
|
