Heat production on SP8 super Z stage

[Jelle Postma]

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Dear all,

I’m very thankful this list exists and have been reading along with great interest! I’d like to discuss a problem that we experience on one of our systems as follows:

- We have a Leica SP8 with The Box/The Cube temperature controlled incubator, Super-Z galvo stage, and CO2/Humidity controlled lid to cover the sample area.

- We notice that the super-z galvo stage becomes noticeably warm to the touch when in use, which affects the samples from our users, who often perform long-term imaging of live plants in petri dishes.

- The local heat at the stage leads to condensation in the sample dishes, drying-out of growing media, and consequentially, unreliable results in the long term imaging experiments.

- Unfortunately, The Box/The Cube cannot counteract the heat development, as the incubator environment is kept at the correct temperature, but the stage itself strongly deviates upward.

So far, we are able to *slightly* alleviate the problem via bottles of ice water, but this is highly variable and short-term. Sub-optimal...

As far as I am aware, there are no super-Z compatible sample-area coolers (but plenty of heaters), and the closest Leica is offering is an objective turret cooler (close, but not the right part)… I am wondering if there are obvious solutions to this that I am missing, whether some of you have found the same problem and possibly an impromptu solution, or whether there are aftermarket parts that counteract this issue.

Thank you very much for your thoughts!

——————

Dr. Jelle Postma
Light microscopy and image analysis
General Instrumentation, FNWI
Radboud University Nijmegen
Huygens Building, Room HG.01.222
Phone: [phone]

[Benjamin Smith]

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If you put galvo stages under load, they will heat-up because they are
using constant current to maintain position.  Perhaps the simplest solution
would be to unplug the galvo when using the incubator and use the motorized
stage/objective turret for XYZ scans (of course, this won't work for XZ
scans).

This is exactly what we did when using a Peltier stage, which is relatively
heavy for a galvo stage.  Speaking of which, Peltier stages are perfect for
both heating and cooling a sample very precisely.  We used this stage for
everything from animal cell culture (37.0 °C) to lamprey embryos (18.0 °C)
and even used it to keep drosophila pupae at exactly 21.0 °C  during a
48-hour live imaging run, to counteract both the temperature swings in the
room and the gradual heating due to the microscope electronics.  This is
the Peltier stage we used: https://physitemp.com/heatcoolstagesmicro_p216

Cheers,
   Ben Smith

On Thu, Mar 18, 2021 at 3:55 AM Jelle Postma <[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.
> *****
>
> Dear all,
>
> I’m very thankful this list exists and have been reading along with great
> interest! I’d like to discuss a problem that we experience on one of our
> systems as follows:
>
> - We have a Leica SP8 with The Box/The Cube temperature controlled
> incubator, Super-Z galvo stage, and CO2/Humidity controlled lid to cover
> the sample area.
>
> - We notice that the super-z galvo stage becomes noticeably warm to the
> touch when in use, which affects the samples from our users, who often
> perform long-term imaging of live plants in petri dishes.
>
> - The local heat at the stage leads to condensation in the sample dishes,
> drying-out of growing media, and consequentially, unreliable results in the
> long term imaging experiments.
>
> - Unfortunately, The Box/The Cube cannot counteract the heat development,
> as the incubator environment is kept at the correct temperature, but the
> stage itself strongly deviates upward.
>
> So far, we are able to *slightly* alleviate the problem via bottles of ice
> water, but this is highly variable and short-term. Sub-optimal...
>
> As far as I am aware, there are no super-Z compatible sample-area coolers
> (but plenty of heaters), and the closest Leica is offering is an objective
> turret cooler (close, but not the right part)… I am wondering if there are
> obvious solutions to this that I am missing, whether some of you have found
> the same problem and possibly an impromptu solution, or whether there are
> aftermarket parts that counteract this issue.
>
> Thank you very much for your thoughts!
>
> ——————
>
> Dr. Jelle Postma
> Light microscopy and image analysis
> General Instrumentation, FNWI
> Radboud University Nijmegen
> Huygens Building, Room HG.01.222
> Phone: [phone]
>

--
Benjamin E. Smith, Ph. D.
Imaging Specialist, Vision Science
University of California, Berkeley
195 Weill Hall
Berkeley, CA  94720-3200
Tel  [phone]
Fax [phone]
e-mail: [hidden email]
https://vision.berkeley.edu/faculty/core-grants-nei/core-grant-microscopic-imaging/

[Craig Brideau]

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I am not sure if you have room, but a water block placed on the stage and
fed with a constant-temperature reservoir of water would let you pull heat
out of the setup. I constructed a Peltier stage for sample heating/cooling
that draws or dumps heat from a water block under it. The working fluid
goes to an external chiller that maintains it at 25 degrees C so the
Peltier modules at the stage level do all the work. Alternatively, you can
forgo the local Peltier modules to save space and just run chilled water
through the block to pull heat out of the area. It will need time to
settle, but once it hits a steady-state temperature it should be quite
resistant to change.

Craig

On Thu, Mar 18, 2021 at 10:43 AM Benjamin Smith <[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.
> *****
>
> If you put galvo stages under load, they will heat-up because they are
> using constant current to maintain position.  Perhaps the simplest solution
> would be to unplug the galvo when using the incubator and use the motorized
> stage/objective turret for XYZ scans (of course, this won't work for XZ
> scans).
>
> This is exactly what we did when using a Peltier stage, which is relatively
> heavy for a galvo stage.  Speaking of which, Peltier stages are perfect for
> both heating and cooling a sample very precisely.  We used this stage for
> everything from animal cell culture (37.0 °C) to lamprey embryos (18.0 °C)
> and even used it to keep drosophila pupae at exactly 21.0 °C  during a
> 48-hour live imaging run, to counteract both the temperature swings in the
> room and the gradual heating due to the microscope electronics.  This is
> the Peltier stage we used: https://physitemp.com/heatcoolstagesmicro_p216
>
> Cheers,
>    Ben Smith
>
> On Thu, Mar 18, 2021 at 3:55 AM Jelle Postma <[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.
> > *****
> >
> > Dear all,
> >
> > I’m very thankful this list exists and have been reading along with great
> > interest! I’d like to discuss a problem that we experience on one of our
> > systems as follows:
> >
> > - We have a Leica SP8 with The Box/The Cube temperature controlled
> > incubator, Super-Z galvo stage, and CO2/Humidity controlled lid to cover
> > the sample area.
> >
> > - We notice that the super-z galvo stage becomes noticeably warm to the
> > touch when in use, which affects the samples from our users, who often
> > perform long-term imaging of live plants in petri dishes.
> >
> > - The local heat at the stage leads to condensation in the sample dishes,
> > drying-out of growing media, and consequentially, unreliable results in
> the
> > long term imaging experiments.
> >
> > - Unfortunately, The Box/The Cube cannot counteract the heat development,
> > as the incubator environment is kept at the correct temperature, but the
> > stage itself strongly deviates upward.
> >
> > So far, we are able to *slightly* alleviate the problem via bottles of
> ice
> > water, but this is highly variable and short-term. Sub-optimal...
> >
> > As far as I am aware, there are no super-Z compatible sample-area coolers
> > (but plenty of heaters), and the closest Leica is offering is an
> objective
> > turret cooler (close, but not the right part)… I am wondering if there
> are
> > obvious solutions to this that I am missing, whether some of you have
> found
> > the same problem and possibly an impromptu solution, or whether there are
> > aftermarket parts that counteract this issue.
> >
> > Thank you very much for your thoughts!
> >
> > ——————
> >
> > Dr. Jelle Postma
> > Light microscopy and image analysis
> > General Instrumentation, FNWI
> > Radboud University Nijmegen
> > Huygens Building, Room HG.01.222
> > Phone: [phone]
> >
>
>
> --
> Benjamin E. Smith, Ph. D.
> Imaging Specialist, Vision Science
> University of California, Berkeley
> 195 Weill Hall
> Berkeley, CA  94720-3200
> Tel  [phone]
> Fax [phone]
> e-mail: [hidden email]
>
> https://vision.berkeley.edu/faculty/core-grants-nei/core-grant-microscopic-imaging/
>

[Jonkman, James]

*****
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*****

Hi, Jelle.  On the plus side, you got a hyperthermia device at no extra charge!

Seriously, though, I agree with Ben that your best bet may be to just unplug it or I might go one step further and remove the Super Galvo Z-stage completely.  This stage looks great on paper but has some pretty serious drawbacks.  We were quoted the galvo stage for an upright multiphoton system, but a mouse is too heavy for this thing (especially with any kind of heating pad to keep the mouse warm).  This was offered as an alternative to a piezo objective positioner that we had asked for in the bid, but we didn't appreciate this key limitation of the weight until it was too late, so now it sits unused in a drawer.  We also received 2 of these galvo stages on inverted microscopes.  On a workhorse confocal (used by many users mostly for fixed cells) we have removed it because it was too easily damaged (replaced twice in the first 6 months or so) and doesn't accommodate easy switching of stage inserts (you have to fiddle with tiny little screws).  And it wasn't really necessary - the built-in focus motor is sufficient for routine applications.  On the other microscope we are still using it, but it's a bit of a pain and I wish we had a piezo instead.  Tokai Hit has a special version of their stage-top incubator that is light enough and fits in the galvo stage, but if you add too much culture medium the weight is too great and the stage starts to squeal.  We haven't noticed hyperthermia but maybe with the galvo stage exposed to the room and just a tiny stage-top incubator the passive heat dissipation is sufficient  - but I'll have to double check the temperature in the coverglass chamber itself now that you've mentioned it.  Good luck and let us know how you solve it!

Cheers,
James

-----------------------------------------------
   James Jonkman, Staff Scientist
   Advanced Optical Microscopy Facility (AOMF)
   and Wright Cell Imaging Facility (WCIF)
   University Health Network
   MaRS, PMCRT tower, 101 College St., Room 15-305
   Toronto, ON, CANADA    M5G 1L7
  [hidden email]  Tel: [phone]
   www.aomf.ca


-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Benjamin Smith
Sent: Thursday, March 18, 2021 12:43 PM
To: [hidden email]
Subject: [External] Re: Heat production on SP8 super Z stage

*****
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*****

If you put galvo stages under load, they will heat-up because they are using constant current to maintain position.  Perhaps the simplest solution would be to unplug the galvo when using the incubator and use the motorized stage/objective turret for XYZ scans (of course, this won't work for XZ scans).

This is exactly what we did when using a Peltier stage, which is relatively heavy for a galvo stage.  Speaking of which, Peltier stages are perfect for both heating and cooling a sample very precisely.  We used this stage for everything from animal cell culture (37.0 °C) to lamprey embryos (18.0 °C) and even used it to keep drosophila pupae at exactly 21.0 °C  during a 48-hour live imaging run, to counteract both the temperature swings in the room and the gradual heating due to the microscope electronics.  This is the Peltier stage we used: https://urldefense.com/v3/__https://physitemp.com/heatcoolstagesmicro_p216__;!!CjcC7IQ!erczknp8X8aXqIu62W0z5PW6sIGafkGHw5hd7oDysrStbX4OaBD3RGd89QqiP50oaxDj$ [physitemp[.]com]

Cheers,
   Ben Smith

On Thu, Mar 18, 2021 at 3:55 AM Jelle Postma <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> https://urldefense.com/v3/__http://lists.umn.edu/cgi-bin/wa?A0=confoca
> lmicroscopy__;!!CjcC7IQ!erczknp8X8aXqIu62W0z5PW6sIGafkGHw5hd7oDysrStbX4OaBD3RGd89QqiP-FO2t5M$ [lists[.]umn[.]edu] Post images on https://urldefense.com/v3/__http://www.imgur.com__;!!CjcC7IQ!erczknp8X8aXqIu62W0z5PW6sIGafkGHw5hd7oDysrStbX4OaBD3RGd89QqiPwbBCSqh$ [imgur[.]com] and include the link in your posting.
> *****
>
> Dear all,
>
> I’m very thankful this list exists and have been reading along with
> great interest! I’d like to discuss a problem that we experience on
> one of our systems as follows:
>
> - We have a Leica SP8 with The Box/The Cube temperature controlled
> incubator, Super-Z galvo stage, and CO2/Humidity controlled lid to
> cover the sample area.
>
> - We notice that the super-z galvo stage becomes noticeably warm to
> the touch when in use, which affects the samples from our users, who
> often perform long-term imaging of live plants in petri dishes.
>
> - The local heat at the stage leads to condensation in the sample
> dishes, drying-out of growing media, and consequentially, unreliable
> results in the long term imaging experiments.
>
> - Unfortunately, The Box/The Cube cannot counteract the heat
> development, as the incubator environment is kept at the correct
> temperature, but the stage itself strongly deviates upward.
>
> So far, we are able to *slightly* alleviate the problem via bottles of
> ice water, but this is highly variable and short-term. Sub-optimal...
>
> As far as I am aware, there are no super-Z compatible sample-area
> coolers (but plenty of heaters), and the closest Leica is offering is
> an objective turret cooler (close, but not the right part)… I am
> wondering if there are obvious solutions to this that I am missing,
> whether some of you have found the same problem and possibly an
> impromptu solution, or whether there are aftermarket parts that counteract this issue.
>
> Thank you very much for your thoughts!
>
> ——————
>
> Dr. Jelle Postma
> Light microscopy and image analysis
> General Instrumentation, FNWI
> Radboud University Nijmegen
> Huygens Building, Room HG.01.222
> Phone: [phone]
>

--
Benjamin E. Smith, Ph. D.
Imaging Specialist, Vision Science
University of California, Berkeley
195 Weill Hall
Berkeley, CA  94720-3200
Tel  [phone]
Fax [phone]
e-mail: [hidden email]
https://urldefense.com/v3/__https://vision.berkeley.edu/faculty/core-grants-nei/core-grant-microscopic-imaging/__;!!CjcC7IQ!erczknp8X8aXqIu62W0z5PW6sIGafkGHw5hd7oDysrStbX4OaBD3RGd89QqiPzm-OBzs$ [vision[.]berkeley[.]edu]

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[Arvydas Matiukas]

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*****

Hi Jelle,

We have a confocal with custom built scanner. The scanner is positioned farther from the microscope stand and warms up (occasionally resulting in nonuniform scan) so within enclosed volume would heat up even more. We found that either switching off the scanner power supply when not scanning and/or after completing the scan returning the  galvos to the position where no current flows through them.

Regarding extra cooling the galvos I would suggest adapting computer CPU liquid cooling system. It can efficiently disperse about 100W of heat and is relatively inexpensive (I very successfully used it to cool custom LED light source).

Best
Arvydas
+++++++++++++++++++++++++++++
Arvydas Matiukas, Ph.D.
Director of  Neuroscience  Microscopy Core
Manager of NRB Shared Research Equipment

SUNY Upstate Medical University
Email: [hidden email]

From: Benjamin Smith<mailto:[hidden email]>
Sent: Thursday, March 18, 2021 12:44 PM
To: [hidden email]<mailto:[hidden email]>
Subject: [EXTERNAL] Re: Heat production on SP8 super Z stage

*****
To join, leave or search the confocal microscopy listserv, go to:
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Post images on https://urldefense.com/v3/__http://www.imgur.com__;!!GobTDDpD7A!YOalK9OFJ1XC1a7vp0Hrr1dDhDb_LERWPmLKR73DngBrtpU1vgTtZ7M4kzCbR-GZJA$<https://urldefense.com/v3/__http:/www.imgur.com__;!!GobTDDpD7A!YOalK9OFJ1XC1a7vp0Hrr1dDhDb_LERWPmLKR73DngBrtpU1vgTtZ7M4kzCbR-GZJA$>  and include the link in your posting.
*****

If you put galvo stages under load, they will heat-up because they are
using constant current to maintain position.  Perhaps the simplest solution
would be to unplug the galvo when using the incubator and use the motorized
stage/objective turret for XYZ scans (of course, this won't work for XZ
scans).

This is exactly what we did when using a Peltier stage, which is relatively
heavy for a galvo stage.  Speaking of which, Peltier stages are perfect for
both heating and cooling a sample very precisely.  We used this stage for
everything from animal cell culture (37.0 °C) to lamprey embryos (18.0 °C)
and even used it to keep drosophila pupae at exactly 21.0 °C  during a
48-hour live imaging run, to counteract both the temperature swings in the
room and the gradual heating due to the microscope electronics.  This is
the Peltier stage we used: https://urldefense.com/v3/__https://physitemp.com/heatcoolstagesmicro_p216__;!!GobTDDpD7A!YOalK9OFJ1XC1a7vp0Hrr1dDhDb_LERWPmLKR73DngBrtpU1vgTtZ7M4kzANpXwZZw$<https://urldefense.com/v3/__https:/physitemp.com/heatcoolstagesmicro_p216__;!!GobTDDpD7A!YOalK9OFJ1XC1a7vp0Hrr1dDhDb_LERWPmLKR73DngBrtpU1vgTtZ7M4kzANpXwZZw$>

Cheers,
   Ben Smith

On Thu, Mar 18, 2021 at 3:55 AM Jelle Postma <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> https://urldefense.com/v3/__http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy__;!!GobTDDpD7A!YOalK9OFJ1XC1a7vp0Hrr1dDhDb_LERWPmLKR73DngBrtpU1vgTtZ7M4kzA6L5p7yg$<https://urldefense.com/v3/__http:/lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy__;!!GobTDDpD7A!YOalK9OFJ1XC1a7vp0Hrr1dDhDb_LERWPmLKR73DngBrtpU1vgTtZ7M4kzA6L5p7yg$>
> Post images on https://urldefense.com/v3/__http://www.imgur.com__;!!GobTDDpD7A!YOalK9OFJ1XC1a7vp0Hrr1dDhDb_LERWPmLKR73DngBrtpU1vgTtZ7M4kzCbR-GZJA$<https://urldefense.com/v3/__http:/www.imgur.com__;!!GobTDDpD7A!YOalK9OFJ1XC1a7vp0Hrr1dDhDb_LERWPmLKR73DngBrtpU1vgTtZ7M4kzCbR-GZJA$>  and include the link in your posting.
> *****
>
> Dear all,
>
> I’m very thankful this list exists and have been reading along with great
> interest! I’d like to discuss a problem that we experience on one of our
> systems as follows:
>
> - We have a Leica SP8 with The Box/The Cube temperature controlled
> incubator, Super-Z galvo stage, and CO2/Humidity controlled lid to cover
> the sample area.
>
> - We notice that the super-z galvo stage becomes noticeably warm to the
> touch when in use, which affects the samples from our users, who often
> perform long-term imaging of live plants in petri dishes.
>
> - The local heat at the stage leads to condensation in the sample dishes,
> drying-out of growing media, and consequentially, unreliable results in the
> long term imaging experiments.
>
> - Unfortunately, The Box/The Cube cannot counteract the heat development,
> as the incubator environment is kept at the correct temperature, but the
> stage itself strongly deviates upward.
>
> So far, we are able to *slightly* alleviate the problem via bottles of ice
> water, but this is highly variable and short-term. Sub-optimal...
>
> As far as I am aware, there are no super-Z compatible sample-area coolers
> (but plenty of heaters), and the closest Leica is offering is an objective
> turret cooler (close, but not the right part)… I am wondering if there are
> obvious solutions to this that I am missing, whether some of you have found
> the same problem and possibly an impromptu solution, or whether there are
> aftermarket parts that counteract this issue.
>
> Thank you very much for your thoughts!
>
> ——————
>
> Dr. Jelle Postma
> Light microscopy and image analysis
> General Instrumentation, FNWI
> Radboud University Nijmegen
> Huygens Building, Room HG.01.222
> Phone: [phone]
>

--
Benjamin E. Smith, Ph. D.
Imaging Specialist, Vision Science
University of California, Berkeley
195 Weill Hall
Berkeley, CA  94720-3200
Tel  [phone]
Fax [phone]
e-mail: [hidden email]
https://urldefense.com/v3/__https://vision.berkeley.edu/faculty/core-grants-nei/core-grant-microscopic-imaging/__;!!GobTDDpD7A!YOalK9OFJ1XC1a7vp0Hrr1dDhDb_LERWPmLKR73DngBrtpU1vgTtZ7M4kzAodu5pkg$<https://urldefense.com/v3/__https:/vision.berkeley.edu/faculty/core-grants-nei/core-grant-microscopic-imaging/__;!!GobTDDpD7A!YOalK9OFJ1XC1a7vp0Hrr1dDhDb_LERWPmLKR73DngBrtpU1vgTtZ7M4kzAodu5pkg$>

[Arvydas Matiukas]

*****
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*****

Hello list,

Did anybody try  the solution from Inscoper “combining hardware and software to control fluorescence microscopes and add-ons easily from one point. … easy-to-use interface that requires no specific software knowledge, and it allows significantly faster image acquisition (3 times more images in the same time than Metamorph/Zen/NIS/LasX and about 5 times more than Micromanager).

Does  there exist  other comparable approaches that are less expensive (this one quotes for academia $25k)

Best,
Arvydas
+++++++++++++++++++++++++++++

Arvydas Matiukas, Ph.D.
Director of  Neuroscience  Microscopy Core
Manager of NRB Shared Research Equipment

SUNY Upstate Medical University

[Csúcs Gábor-3]

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Dear Arvydas,

Yes, we have quite extensively tested the Inscoper "box" and we are planning to purchase it for one (but only one) of our systems. Concerning your question and the advertised speed gain I can't fully confirm those values. There might be some cases where such a speed difference can be observed but this depends very much on the actual system/experiment and I could tell you also experiments where there will be no speed gain at all.
What Inscoper does, is similar to what numerous microscope companies have already done (but only for their dedicated hardware) while Inscoper can offer it (theoretically) for any component. The main idea is a hardware trigger/control box that can function independently on your acquisition computer, hence eventual other computational processes will not slow down you acquisition, you should be able to use your hardware at the maximum (physically possible) speed. If you have an acquisition sequence with lot of hardware control (e.g. filter wheels etc.) the speed gain can be indeed significant. A regular external filter wheel has approximately 50 ms switching time between neighboring position but due to software issues you will probably switch it with a speed of 150-200ms when you control it directly via your computer. With the Inscoper box you should reach the 50ms theoretical limit. On the other hand, if you use your camera with a software without any hardware control (e.g. stream acquisition) there will be no speed gain with Inscoper as the camera is in any case the limiting factor.
        What speaks against Inscoper (except that you need to pay for it)? They are a small company with limited software development possibilities, so I would claim that the flexibility of their software does not reach the level of e.g. NIS Elements.

Greetings GAbor




-----Original Message-----
From: Confocal Microscopy List <[hidden email]> On Behalf Of Arvydas Matiukas
Sent: Friday, March 19, 2021 7:47 PM
To: [hidden email]
Subject: Speeding up camera acquisition 3x

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Hello list,

Did anybody try  the solution from Inscoper “combining hardware and software to control fluorescence microscopes and add-ons easily from one point. … easy-to-use interface that requires no specific software knowledge, and it allows significantly faster image acquisition (3 times more images in the same time than Metamorph/Zen/NIS/LasX and about 5 times more than Micromanager).

Does  there exist  other comparable approaches that are less expensive (this one quotes for academia $25k)

Best,
Arvydas
+++++++++++++++++++++++++++++

Arvydas Matiukas, Ph.D.
Director of  Neuroscience  Microscopy Core Manager of NRB Shared Research Equipment

SUNY Upstate Medical University