Imaging at below room temperature

[Laura Macro]

Dear all,

We are trying to do some imaging of yeast and Dictyostelium and have found that our microscope cubicles are around 24-25C which is affecting the experiments.  Ideally we would like to image at around 22C or other temperatures below room temperature.  I was wondering if people could let me know if they have any solutions to this problem.  Our microscopes are sealed (perspex boxes) and we have heaters which work great for imaging at 37C.  

Thanks

Laura Macro

[PattyJansma]

Hi, Laura,

     When imaging insect cell cultures, we needed stable temperature
control at 25C. We are using a Harvard Apparatus Temperature Controller
TC-202A and it has worked very well for several years.


Patty

Patty Jansma
ARLDN
University of AZ
Tucson, AZ

At 09:53 AM 11/17/2008, you wrote:

>Dear all,
>
>We are trying to do some imaging of yeast and Dictyostelium and have found
>that our microscope cubicles are around 24-25C which is affecting the
>experiments.  Ideally we would like to image at around 22C or other
>temperatures below room temperature.  I was wondering if people could let
>me know if they have any solutions to this problem.  Our microscopes are
>sealed (perspex boxes) and we have heaters which work great for imaging at
>37C.
>
>Thanks
>
>Laura Macro

[Julio Vazquez]

Hi Laura, 

We had similar issues with people imaging live C. elegans. Worms are pretty sensitive to temperature. We managed to keep them happy by wrapping a coil of copper tubing around the 40x oil objective and circulating water chilled to about 8-10 degrees centigrade through it with a Lauda refrigerating water bath. This cools down the objective, and by contact, also the sample, to about 16-18 degrees centigrade.

More sophisticated perhaps, some vendors have Peltier cooled or water cooled stage inserts; check for instance:

www.instec.com

http://www.20-20tech.com/

When getting some of the cooling stage inserts, you need to make sure they will work for your particular sample. For instance, the one we have from 20-20 (bionomic system) will not work with high NA immersion objectives, because the metal plate that holds the sample is too thick. We can make it work though by gluing a coverslip UNDER the insert with oil, and placing the sample on the coverslip.

--

Julio Vazquez

Fred Hutchinson Cancer Research Center

1100 Fairview Ave. N.,  mailstop DE-512

Seattle, WA 98109-1024

[hidden email]

http://www.fhcrc.org/science/shared_resources/imaging/

On Nov 17, 2008, at 8:53 AM, Laura Macro wrote:

Dear all,

We are trying to do some imaging of yeast and Dictyostelium and have found that our microscope cubicles are around 24-25C which is affecting the experiments.  Ideally we would like to image at around 22C or other temperatures below room temperature.  I was wondering if people could let me know if they have any solutions to this problem.  Our microscopes are sealed (perspex boxes) and we have heaters which work great for imaging at 37C.  

Thanks

Laura Macro

[Knecht, David]

We have a 20/20 Technologies stage temp controller (http://www.20-20tech.com/)  that we have used for this.  THermal transfer to plastic is not great and so it is hard to determine the exact temperature at the image plane, but we have used a IR thermometer to estimate.  Another peltier system I know of is http://www.alascience.com/.  Dave

On Nov 17, 2008, at 12:39 PM, Julio Vazquez wrote:

Hi Laura, 

We had similar issues with people imaging live C. elegans. Worms are pretty sensitive to temperature. We managed to keep them happy by wrapping a coil of copper tubing around the 40x oil objective and circulating water chilled to about 8-10 degrees centigrade through it with a Lauda refrigerating water bath. This cools down the objective, and by contact, also the sample, to about 16-18 degrees centigrade.

More sophisticated perhaps, some vendors have Peltier cooled or water cooled stage inserts; check for instance:

www.instec.com

http://www.20-20tech.com/

When getting some of the cooling stage inserts, you need to make sure they will work for your particular sample. For instance, the one we have from 20-20 (bionomic system) will not work with high NA immersion objectives, because the metal plate that holds the sample is too thick. We can make it work though by gluing a coverslip UNDER the insert with oil, and placing the sample on the coverslip.

--

Julio Vazquez

Fred Hutchinson Cancer Research Center

1100 Fairview Ave. N.,  mailstop DE-512

Seattle, WA 98109-1024

[hidden email]

http://www.fhcrc.org/science/shared_resources/imaging/

On Nov 17, 2008, at 8:53 AM, Laura Macro wrote:

Dear all,

We are trying to do some imaging of yeast and Dictyostelium and have found that our microscope cubicles are around 24-25C which is affecting the experiments.  Ideally we would like to image at around 22C or other temperatures below room temperature.  I was wondering if people could let me know if they have any solutions to this problem.  Our microscopes are sealed (perspex boxes) and we have heaters which work great for imaging at 37C.  

Thanks

Laura Macro

Dr. David Knecht    

Department of Molecular and Cell Biology

Co-head Flow Cytometry and Confocal Microscopy Facility

U-3125

91 N. Eagleville Rd.

University of Connecticut

Storrs, CT 06269

860-486-2200

860-486-4331 (fax)

[Michael Weber-4]

Laura,

LIS (Life Imaging Services) offers a cage incubator with cooling module
(external heat exchanger). Since you already have the boxes, maybe they
can fit the external module to yours. I just saw a demo of the unit and
never tested it in real life, but something like this is probably the
most stable solution.

Another option would be a stage incubator for cooling, like the TB-3
from Warner Instruments (Peltier system) or the Delta T from Bioptechs
(water-cooling solution). Disadvantages: temperature at the sample
depends more on the room temperature, immersion objectives bring heat in
and need to be cooled separately.

No connection to the mentioned companies.

Michael


Laura Macro wrote:

> Dear all,
>
> We are trying to do some imaging of yeast and Dictyostelium and have
> found that our microscope cubicles are around 24-25C which is affecting
> the experiments.  Ideally we would like to image at around 22C or other
> temperatures below room temperature.  I was wondering if people could
> let me know if they have any solutions to this problem.  Our microscopes
> are sealed (perspex boxes) and we have heaters which work great for
> imaging at 37C.  
>
> Thanks
>
> Laura Macro

[Dale Callaham]

Hi Laura,

You could install Peltier coolers to cool the whole box. If you have
anyone handy with simple electronics these Peltier coolers would
probably drop the temp inside your plexiglass box a few degrees....

http://www.allelectronics.com/make-a-store/item/PJT-10/LARGE-THERMOELECTRIC-DEVICE/1.html

Cheers!

Dale

>>
>>
>>
>> On Nov 17, 2008, at 8:53 AM, Laura Macro wrote:
>>
>>> Dear all,
>>>
>>> We are trying to do some imaging of yeast and Dictyostelium and have
>>> found that our microscope cubicles are around 24-25C which is
>>> affecting the experiments.  Ideally we would like to image at around
>>> 22C or other temperatures below room temperature.  I was wondering if
>>> people could let me know if they have any solutions to this problem.  
>>> Our microscopes are sealed (perspex boxes) and we have heaters which
>>> work great for imaging at 37C.  
>>>
>>> Thanks
>>>
>>> Laura Macro
>>
>

[Dan Focht]

Laura

If you intend to image below room temperature we have several options.

With yeast, you undoubtedly will be using high mag, high N.A. objectives so cooling the objective is a must.

However if there is too much of a temperature difference condensation can form on the optic at the nosepiece end of the objective.

Bioptechs makes a thermal isolator that you screw the objective into then screw the objective with the isolator into the nosepiece.  

This provides a twofold advantage.

1. It isolates the nosepiece from the objective thereby increasing the efficiency of thermal transfer to a cooling means.

2. It provides a gas trap that can be filled with dry air (not Nitrogen due to its RI) to prevent condensation from forming.

Bioptechs also makes custom Cooling rings to precisely fit the objective to maximize the transfer of heat from the objective.

I have a few questions about your setup.

Are you using an upright or inverted scope?

What modes of microscopy are you using?

What is the N.A. of your condenser if you are using transmitted light?

Is the speed of temperature transition of concern?

Additional micro-environmental control systems for cooling and warming are available at our web site www.bioptechs.com

Dan

On Nov 17, 2008, at 11:53 AM, Laura Macro wrote:

Dear all,

We are trying to do some imaging of yeast and Dictyostelium and have found that our microscope cubicles are around 24-25C which is affecting the experiments.  Ideally we would like to image at around 22C or other temperatures below room temperature.  I was wondering if people could let me know if they have any solutions to this problem.  Our microscopes are sealed (perspex boxes) and we have heaters which work great for imaging at 37C.  

Thanks

Laura Macro

Dan Focht

Bioptechs

724-282-7145

www.bioptechs.com

[Beat Ludin]

Dear Laura

As Michael already pointed out, we offer the
iceCUBE&BOX temperature control system which
covers a range of ambient-10°C to 45°C for most
setups. One of the advantages of an integral
system over a local one is that it will work with
any sample, stage and objective. Don't hesitate
to contact me off-list if you have further questions.

It goes without saying that I have considerable
commercial interest in our own products :-)

Best regards,

Beat

At 17:53 17-11-2008, you wrote:

>Dear all,
>
>We are trying to do some imaging of yeast and
>Dictyostelium and have found that our microscope
>cubicles are around 24-25C which is affecting
>the experiments.  Ideally we would like to image
>at around 22C or other temperatures below room
>temperature.  I was wondering if people could
>let me know if they have any solutions to this
>problem.  Our microscopes are sealed (perspex
>boxes) and we have heaters which work great for imaging at 37C.
>
>Thanks
>
>Laura Macro

[Ian Dobbie]

Dan Focht <[hidden email]> writes:

> 2. It provides a gas trap that can be filled with dry air (not Nitrogen due to
> its RI) to prevent condensation from forming.

The RI difference between air and nitrogen is < 1:10^4 so this
seems to be totally unnecessary.

Ian