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 |
In reply to this post by Laura Macro
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 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 On Nov 17, 2008, at 8:53 AM, Laura Macro wrote: Dear all, |
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:
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 |
In reply to this post by Laura Macro
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 |
In reply to this post by Knecht, David
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 >> > |
In reply to this post by Laura Macro
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, Dan Focht Bioptechs 724-282-7145 www.bioptechs.com |
Beat Ludin |
In reply to this post by Laura Macro
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 |
In reply to this post by Dan Focht
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 |
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