Incubation. Cage or Stage

[Andrew Vaughan-2]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Dear All,

In common with many of you, I manage a facility where users want to use many different model organisms. We mostly have cage type incubation systems on our microscopes, which is less than ideal when one user wants the microscope at 26 degrees C and the next user wants it at 37 degrees C. I have found that the cage-type incubators do have the advantage of suppressing temperature fluctuation caused by thermostat control AC, so it's good to have them around the machine. However, I would like to know whether there are stage type incubation systems I could use inside the chambers that would allow us to quickly switch between imaging specimens at 20, 26 and 37 degrees C (possibly below ambient, so with cooling) but that provide a reliable way of maintaining focus with high NA immersion lenses. I have used objective lens heaters before and some (but not all) have been very good, but are there any that won't clash with the stage or microscope when the nose piece is rotated? I'd like to leave them in position rather than have to take them off all the time. Any suggestions would be most welcome.

Thanks,

Yours faithfully,

Andrew Vaughan.

[Darran Clements]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Hi Andrew,
We use hotbox type boxes too, but we have an LSM 710 with a Pecon stagetop incubation system for Ibidi microwell plates that rapidly equalises to the required temperature. It has excellent XY repositioning and the Zen autofocus works well for slower time lapse experiments, though a Definite focus/Perfect focus/ZDC type hardware autofocusing might be a faster means of getting you back to a stable z position.

Hotboxes are great, but the expansion and contraction time for a stable microscope setup under different conditions is tedious, the stagetop design works reasonably well to fill the need you've detailed for our needs.

All the best
Darran Clements
Stem Cell Institute
Cambridge

Get Outlook for Android<https://aka.ms/ghei36>

________________________________
From: Confocal Microscopy List <[hidden email]> on behalf of Andrew Vaughan <[hidden email]>
Sent: Monday, July 13, 2020 4:31:17 PM
To: [hidden email] <[hidden email]>
Subject: Incubation. Cage or Stage

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Dear All,

In common with many of you, I manage a facility where users want to use many different model organisms. We mostly have cage type incubation systems on our microscopes, which is less than ideal when one user wants the microscope at 26 degrees C and the next user wants it at 37 degrees C. I have found that the cage-type incubators do have the advantage of suppressing temperature fluctuation caused by thermostat control AC, so it's good to have them around the machine. However, I would like to know whether there are stage type incubation systems I could use inside the chambers that would allow us to quickly switch between imaging specimens at 20, 26 and 37 degrees C (possibly below ambient, so with cooling) but that provide a reliable way of maintaining focus with high NA immersion lenses. I have used objective lens heaters before and some (but not all) have been very good, but are there any that won't clash with the stage or microscope when the nose piece is rotated? I'd like to leave them in position rather than have to take them off all the time. Any suggestions would be most welcome.

Thanks,

Yours faithfully,

Andrew Vaughan.

[Jens Breffke]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

*** COMMERCIAL RESPONSE ***

Hi Andrew,

We carry a brand new product by a startup company called Linnowave which is out of the Max-Planck Institute in Erlangen, Germany. Their novel solution is to have the actual cover slip being the heating and measuring element using transparent, conductive coatings. It is measuring and generating heat where you need it: near the focal plane at the cover slip surface! I comes as a 18x18mm cover slip size and is available also with a PDMS liquid reservoir. All you need a humidity chamber which can be equilibrated much faster than conventional heating elements based on equilibration of an entire sample system, including objective. Since the temperature control with the Linnowave VAHEAT is targeted at the very volume where your focus is it can adapt within seconds.
For more information please visit  https://linnowave.com/
and https://shop.boselec.com/products/vaheat-microscope-temperature-control

With best regards from Boston, MA,

-Jens
____
Jens Breffke, PhD
Boston Electronics
[phone]
[hidden email]
www.boselec.com
 

-----Original Message-----
From: Confocal Microscopy List <[hidden email]> On Behalf Of Andrew Vaughan
Sent: Monday, July 13, 2020 11:31 AM
To: [hidden email]
Subject: Incubation. Cage or Stage

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Dear All,

In common with many of you, I manage a facility where users want to use many different model organisms. We mostly have cage type incubation systems on our microscopes, which is less than ideal when one user wants the microscope at 26 degrees C and the next user wants it at 37 degrees C. I have found that the cage-type incubators do have the advantage of suppressing temperature fluctuation caused by thermostat control AC, so it's good to have them around the machine. However, I would like to know whether there are stage type incubation systems I could use inside the chambers that would allow us to quickly switch between imaging specimens at 20, 26 and 37 degrees C (possibly below ambient, so with cooling) but that provide a reliable way of maintaining focus with high NA immersion lenses. I have used objective lens heaters before and some (but not all) have been very good, but are there any that won't clash with the stage or microscope when the nose piece is rotated? I'd like to leave them in position rather than have to take them off all the time. Any suggestions would be most welcome.

Thanks,

Yours faithfully,

Andrew Vaughan.

[weber]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Hi Andrew,

What kind of microscope did you use ? I think it is an interted one but in which conditions ?

Regards

[Andrew Vaughan-2]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

The microscopes are inverted. The rooms are just standard microscope laboratories. The conditions vary according to the model organism. Some users have mammalian cells which need 37 degrees C, some have yeast which are happy around 20 degrees C and some people are using Drosophila tissue at 26 degrees C.

[Jeremy Adler-4]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Hi Andrew,

Having looked at your literature, I am not clear where the temperature is measured.
Is an integrated measurement across the whole coverslip being made?
If so any immersion objective will act as a heat sink and reduce the local temperature for the actual cells being imaged, but this will contribute little to an overall temperature measurement - and not be compensated. If compensated and the heating increased, then cells at a distance from the objective will be heated above 37.

I have a strong preference for heating everything to 37, then local gradients around the cells/objective cease to be an issue.

Have a look at J Microscopy 210 (2003) 131-137,
We found some wonderful distortions when a slow Z series had a similar timescale to the room temperature oscillations.

Jeremy
===============================================
                    B i o V i s   P l a t f o r m of  Uppsala University
                   Light & EM microscopy / FlowCytometry & Cell Sorting / Image Analysis
===============================================
Jeremy Adler   PhD - Senior research engineer
Light, Confocal Microscopy, Image Analysis
E-mail: [hidden email]
070-1679349

Dag Hammarskjölds v 20
751 85 UPPSALA, SWEDEN
http://biovis.uu.se/
===============================================











-----Original Message-----
From: Confocal Microscopy List <[hidden email]> On Behalf Of Jens Breffke
Sent: Monday, July 13, 2020 6:06 PM
To: [hidden email]
Subject: Re: Incubation. Cage or Stage

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

*** COMMERCIAL RESPONSE ***

Hi Andrew,

We carry a brand new product by a startup company called Linnowave which is out of the Max-Planck Institute in Erlangen, Germany. Their novel solution is to have the actual cover slip being the heating and measuring element using transparent, conductive coatings. It is measuring and generating heat where you need it: near the focal plane at the cover slip surface! I comes as a 18x18mm cover slip size and is available also with a PDMS liquid reservoir. All you need a humidity chamber which can be equilibrated much faster than conventional heating elements based on equilibration of an entire sample system, including objective. Since the temperature control with the Linnowave VAHEAT is targeted at the very volume where your focus is it can adapt within seconds.
For more information please visit  https://linnowave.com/ and https://shop.boselec.com/products/vaheat-microscope-temperature-control

With best regards from Boston, MA,

-Jens
____
Jens Breffke, PhD
Boston Electronics
[phone]
[hidden email]
www.boselec.com


-----Original Message-----
From: Confocal Microscopy List <[hidden email]> On Behalf Of Andrew Vaughan
Sent: Monday, July 13, 2020 11:31 AM
To: [hidden email]
Subject: Incubation. Cage or Stage

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Dear All,

In common with many of you, I manage a facility where users want to use many different model organisms. We mostly have cage type incubation systems on our microscopes, which is less than ideal when one user wants the microscope at 26 degrees C and the next user wants it at 37 degrees C. I have found that the cage-type incubators do have the advantage of suppressing temperature fluctuation caused by thermostat control AC, so it's good to have them around the machine. However, I would like to know whether there are stage type incubation systems I could use inside the chambers that would allow us to quickly switch between imaging specimens at 20, 26 and 37 degrees C (possibly below ambient, so with cooling) but that provide a reliable way of maintaining focus with high NA immersion lenses. I have used objective lens heaters before and some (but not all) have been very good, but are there any that won't clash with the stage or microscope when the nose piece is rotated? I'd like to leave them in position rather than have to take them off all the time. Any suggestions would be most welcome.

Thanks,

Yours faithfully,

Andrew Vaughan.








När du har kontakt med oss på Uppsala universitet med e-post så innebär det att vi behandlar dina personuppgifter. För att läsa mer om hur vi gör det kan du läsa här: http://www.uu.se/om-uu/dataskydd-personuppgifter/

E-mailing Uppsala University means that we will process your personal data. For more information on how this is performed, please read here: http://www.uu.se/en/about-uu/data-protection-policy

[Pierre Tuerschmann]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

*** COMMERCIAL RESPONSE ***

Dear Jeremy and all,

Let me help to clarify: Our system VAHEAT is specifically designed to compensate the effects your were mentioning:
1) Local heat sink in the FOV
2) thermal drifts.

We have a highly sensitive temperature probe sitting in the FOV that enables fast feedback to adjust for temperature variations in your environment. The specific geometrical arrangement of our heating element, temperature probe and the immersion objective ensures a homogene temperature distribution in the FOV. Additionally, the heat load injected into your objective via the immersion oil is kept to a minimum and the quality of the optical imaging system can be maintained even at elevated temperatures (up to 80°C). This allows for fast, reproducible temperature changes and long-term temperature stability without warming the whole microscope to your desired temperature (which takes several minutes to hours and usually doesn’t improve on the temperature accuracy). All in all VAHEAT can, thus, be combined with various highly sensitive measurements such as AFM/confocal, STORM or PALM, PAINT, STED, TIRF.

For further details feel free to reach out to us. We are always happy to discuss your specific requirements.

Best regards from Erlangen

Pierre
———————————————
Dr. Pierre Türschmann
Managing Director

Linnowave GmbH
Henkestrasse 91, D-91052 Erlangen
Phone [phone]
E-mail [hidden email]
Web https://linnowave.com

Registergericht Fürth HRB 17123
USt-IdNr. DE 320927905

[Dan Focht]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Andrew

Here is how I address the micro-environmental issues you described. I agree the box approach does reduce the thermal expansion of the whole scope that occurs due to room temp variations. However in that you describe the need to use immersion lenses for and during temperature transitions I recommend keeping the thermal mass of the specimen containment structure to a minimum and applying or removing heat to that immediate area in the most efficient means possible. Waiting for self equilibration takes a loooong time. My company, Bioptechs, has developed  micro-environmental control systems for both steady state and quick thermal ramping applications such as gene expression experiments for 28 years. Therefore, I believe we can precisely  meet your needs. In addition to typical fixed physiological temperatures, our systems are being used in the temperature ranges of, 15C to 25C in 20 seconds, 4C to 40C in 3 minutes, 15C to 60C in 2 minutes while maintaining stability in the Z axis within our FCS2 flow cell. If you don't need an actual laminar flow cell we have a number of other products and technologies for fast and efficient thermal control as well as Z axis stability to choose from including a coverslip bottomed dish with intrinsic temperature control at the coverslip. Temperature control for the objective is a specialty of ours. Our patented Objective Heater incorporates the objective temperature at the specimen plane in the control loop and compensates for the heat-sinking effect of the nosepiece. It has a detachable cable from the heater so that you can rotate the nosepiece. If you need to obtain temperatures below ambient we have products to do that as well. Special note, we developed a thermal control system used on all our products that takes the specimen from initial to target temperature typically in seconds then steady thereafter and it does not induce the variations typical with PID. Setting up a micro-environmental control system for imaging is not trivial. Due to the number of technical details involved in properly defining an environmental system, I would like you to call me. Prior to calling it would be beneficial to visit our website to see the products and technologies available. www.bioptechs.com.


Dan Focht
Bioptechs Inc.
3560 Beck Road
Butler, PA 16002-9259
Office: 724-282-7145
Toll Free: 877-LIVE-CELL (548-3235)
[hidden email]
www.bioptechs.com


On Jul 14, 2020, at 4:56 AM, Andrew Vaughan <[hidden email]> wrote:

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

The microscopes are inverted. The rooms are just standard microscope laboratories. The conditions vary according to the model organism. Some users have mammalian cells which need 37 degrees C, some have yeast which are happy around 20 degrees C and some people are using Drosophila tissue at 26 degrees C.