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Hello, I´m going to build a heating stage/chamber in
order to do live imaging at 37°C with the ordinary 35mm-dishes . Due to lack of
experience with these systems are there any general advices about eventual
pitfalls or what works perfectly fine/not? Are there eventually some instruction
guidelines out there? Since I´ve built a complete home-made
scope, I´m not particularly limited in terms of space or compatibility to
Zeiss, Olympus, Leica etc. Currently, I´ve two separate systems in mind: 1)
A flexible system that separately
warms the objective as well as the dish. -
Does that work efficiently or
is the loss of heat too significant rendering this method rather meaningless? -
Any recommendations where to
get such warming equipment for reasonable prices? 2)
A complete plexiglas chamber
which could also be utilized for CO2-adjustment. -
Are these incubation chambers so
relatively big volume-wise because of trying to avoid significant flows which
could disturb the sample? Or is it of other practical reasons? -
What kind of heating source (ventilator)
would you suggest, any particular recommendations? Many thanks in advance for taking the time,
I´m happy about any advice. Best regards, Steffen Steffen Steinert, Dipl.-Ing. -------------------------------------- Universität
Stuttgart 3.
Physikalisches Institut Pfaffenwaldring
57 70550
Stuttgart Tel.:
049/0711/68569832 Fax:
049/0711/68565281 http://www.pi3.uni-stuttgart.de/en/ -------------------------------------- |
Hugo.Ostermann |
Search the CONFOCAL archive at
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Dear Steffen, you should see
bioptechs.com. The have a very detailed catalogue and movies with instructions
on the net. We have a complete
demo-kit if you would be interested to test their equipment. Just contact me
off-list. Mit freundlichen Grüßen, Hugo Ostermann CHROMAPHOR
Analysen-Technik GmbH Mühlenkamp 37 D-59387 Ascheberg - Germany Tel: ++49 - 2593-928.600 Fax: ++49 -
2593-928.601 mail: URL:
http://www.chromaphor.de Sitz Ascheberg-Westfalen Amtsgericht - Registergericht Coesfeld HRB
7703 Geschäftsführer: Hugo Ostermann Von: Hello, I´m going to build a heating stage/chamber in order to do live imaging
at 37°C with the ordinary 35mm-dishes . Due to lack of experience with these
systems are there any general advices about eventual pitfalls or what works
perfectly fine/not? Are there eventually some instruction guidelines out there? Since I´ve built a complete home-made scope, I´m not particularly
limited in terms of space or compatibility to Zeiss, 1) A flexible system that separately warms the objective as well as the
dish. -
Does that work
efficiently or is the loss of heat too significant rendering this method rather
meaningless? -
Any
recommendations where to get such warming equipment for reasonable prices? 2) A complete plexiglas chamber which could also be utilized for
CO2-adjustment. -
Are these
incubation chambers so relatively big volume-wise because of trying to avoid
significant flows which could disturb the sample? Or is it of other practical
reasons? -
What kind of
heating source (ventilator) would you suggest, any particular recommendations? Many thanks in advance for taking the time, I´m happy about any advice. Best
regards, Steffen Steffen Steinert, Dipl.-Ing. -------------------------------------- Universität Stuttgart 3. Physikalisches Institut Pfaffenwaldring 57 70550 Stuttgart Tel.: 049/0711/68569832 Fax: 049/0711/68565281 http://www.pi3.uni-stuttgart.de/en/ -------------------------------------- |
In reply to this post by Steffen Steinert
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
I am currently in the same task, for our confocal (that already is
surrounded by an environmental chamber "the box") but after asking a lot of
experienced people;
Thinking in the cost-benefice of a commercial chamber (and controllers)
versus a custom-made-by-yourself chamber:
-Cost ($) probably will be around the same range
-In the custom-made-by-yourself you do not have guarantee...
If you want something special, you can ask Warner, Oko-Lab or Bioptechs for
custom made chambers.
these are some useful links, depending on your purpose:
?
FIRST: All-in-one solutions for Microscope Stage Incubator
(especially for multiwell plates as
http://www.glassbottomdishes.com/multiwellproducts.html)
-Incubator in a plate enabling time lapse imaging
(http://www.wafergen.com/slide50.html)
-Just one incubator for every multiwell plate
(http://www.oko-lab.com/88.page, for USA through Warner
http://www.warneronline.com/product_info.cfm?id=1228α=a&name=Oko%20Labs%20Microscope%20Stage%20Incubator#)
?
SECOND: Separated components for Microscope Stage Incubator:
-Closed live-cell micro-observation chamber
(http://www.bioptechs.com/Products/FCS2/fcs2.html) + Objective Heater System
(http://www.bioptechs.com/Products/OBJ_HTR/obj_htr.html)
-Confocal Imaging Chambers
(http://www.warneronline.com/product_info.cfm?id=774α=a&name=RC%2D30%2C%20RC%2D30HV%2C%20and%20RC%2D30WA%20Confocal%20Imaging%20Chambers)
-Culture dish holder
(http://www.warneronline.com/product_info.cfm?name=DH-35%20Culture%20Dish%20Heater&id=766)
?
THIRD: If we want to perform micromanipulation or microinjection
-Open Dish System (http://www.bioptechs.com/Products/Delta_T/delta_t.html)
?
That's all, I am sorry for the long URL's
I hope this would be useful,
Fernando
--
Fernando Avila-Rencoret, MD
Department of Biomedical Engineering
Carnegie Mellon University
***************************************************
PS: An in-deep review on the topic:
http://www.microscopyu.com/articles/livecellimaging/culturechambers.html
|
This post was updated on .
In reply to this post by Steffen Steinert
Dear Steffen,
amongst many other commercial suppliers for heating equipment, EMBL sells controllers or complete mic enclosures, and as far as I know the only system to control CO2 in the entire mic enclosure. The system is meanwhile widely used. I have been involved with the development of the system (I declare a minimal commercial interest, I receive some inverntor compensation, though almost the entire money is reinvested into development of technology and support of the workshops). Concerning the objective heater solution, these put serious stress on the objectives and the heated volume is small, so to produce a long term stable situation, its not ideal. However, for fast heating and cooling, like for example to switch on/off temp sensitve allels, these heaters are suitable. EMBL has developed a device to heat/cool a small volume at 1degC per sec. For more info on enclosure heater+CO2 or the fast response heater please take a look at http://www.embl-em.de/products/ The product IDs are 159 & 396 Cheers, jens ***Join the elmi meeting in Davos! More info: http://elmi08.unibas.ch/index.html ***
|
In reply to this post by Steffen Steinert
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Steffen Steinert <[hidden email]> writes: > 1) A flexible system that separately warms the objective as well as the > dish. > > - Does that work efficiently or is the loss of heat too significant > rendering this method rather meaningless? If you have immersion objectives then you must also warm the objective, as you seem to realise. The heaters are rather harsh on objectives, which are not really designed for thermal stresses, especially if you want rapid heating or cooling. You also end up with a pretty dramatic gradient from the heating region on the objective into the microscope nose cone. The main advantage of this type of setup is that rapid changes in temperature are possible. > 2) A complete plexiglas chamber which could also be utilized for > CO2-adjustment. > > - Are these incubation chambers so relatively big volume-wise because > of trying to avoid significant flows which could disturb the sample? Or is it > of other practical reasons? My experience is that this type of chamber is necessary to get good long term stability of the microscope. If you are doing time lapse of more than a few mins then temperature drift can shift both the x-y position and probably more significantly the focus. Air conditioning is particularly bad for inducing cyclical temperature changes of a few degrees around the "set" temperature. But any room is going to warm up during the day and cool down at night. > - What kind of heating source (ventilator) would you suggest, any > particular recommendations? The equipment I have used was all custom built but broadly consisted of a fan running continuously and a heater that is pulsed by a PID controller. You setup the system, then let the PID auto-tune its control parameters to maintain the temp that you want. Works very well. Ian |
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Steffen I apologize for not getting back to you sooner. I hope this message saves you a lot of time and effort. Here are a few pointers. 1) A flexible system that separately warms the objective as well as the dish. It is important that the two systems do not conflict with each other. The objective warmers job is to prevent heat loss from the specimen only. It should not be used to warm the specimen. That is the job of the specimen warmer. A properly engineered objective heater system will not damage the objective. I have thermographed dozens of objectives to characterize their thermal profile when heated and cooled. It is important to know how an objective behaves. Once you know how heat propagates through the objective it is easy to thermal regulate it. If you don't know its thermal characteristics you are shooting in the dark and subject to making mistakes. That is why I posted this information on my web site. I have investigated this subject for fifteen years and post the results on our web page complete with descriptions, control algorithm, pictures of many brands and types of objectives, thermographs, time-lapse thermographs both with and without thermal isolation of the objective from the nosepiece. http:// www.bioptechs.com/Products/OBJ_HTR/obj_htr.html Please refer to this website and if you have any further questions don't hesitate to contact me. As for an economical way of warming an ordinary 35mmm culture dish, please check out the Bioptechs Stable Z system. I believe you will find that it is the only dish warming system on the market that makes sense in its design. It is NOT just a plate of metal with a heater on it like so many other "stage warmers". It provides peripheral thermal support to any 35mm dish without inducing Z axis drift. You will also find that it cost less than the traditional "stage heating" systems that are common on the market today. There is a detailed diagram on our website explaining how it works. There you go, same heat, less cost, better performance. http://www.bioptechs.com/ Products/Stable_Z/Stable_Z.html 2) A complete plexiglas chamber which could also be utilized for CO2-adjustment. We do not support boxing the microscope, instead we show you how you can set up low volume ,low consumption, localized and humidified CO2 environments for your specimen without investing in the box. You will find many other alternative methods for temperature, CO2, and perfusion control as well as techniques and equipment to accommodate a variety of specimen types. www.bioptechs.com Dan Content-Type: text/html Search the CONFOCAL archive at http:// listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Hello, I´m going to build a heating stage/chamber in order to do live imaging at 37°C with the ordinary 35mm-dishes . Due to lack of experience with these systems are there any general advices about eventual pitfalls or what works perfectly fine/not? Are there eventually some instruction guidelines out there? Since I´ve built a complete home-made scope, I´m not particularly limited in terms of space or compatibility to Zeiss, Olympus, Leica etc. Currently, I´ve two separate systems in mind: 1) A flexible system that separately warms the objective as well as the dish. - Does that work efficiently or is the loss of heat too significant rendering this method rather meaningless? - Any recommendations where to get such warming equipment for reasonable prices? 2) A complete plexiglas chamber which could also be utilized for CO2-adjustment. - Are these incubation chambers so relatively big volume- wise because of trying to avoid significant flows which could disturb the sample? Or is it of other practical reasons? - What kind of heating source (ventilator) would you suggest, any particular recommendations? Many thanks in advance for taking the time, I´m happy about any advice. Best regards, Steffen Steffen Steinert, Dipl.-Ing. Dan Focht Bioptechs 3560 Beck Rd. Butler, PA 16002 V724-282-7145 F724-282-0745 Micro-Environmental Control Systems www.bioptechs.com |
In reply to this post by Ian Dobbie-2
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear all, Thanks for the input! Although it requires some more time, I decided to build a complete heating chamber by myself, which so far turned out to be rather cheap (all items ~400Euro). Now it´s just a matter of putting it all together... Once again, thanks for the advices, particular thanks to Michael Herron for very helpful practical hints! Cheers, Steffen |
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