avoid condensation??

[Christian Tischer]

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Hello,

we are using an inverted microscope and need to do long term transmission
imaging in a 24-well dish. the whole central part of microscope is surrounded by
a box keeping it at 37 degrees and providing CO2.

so far, we fail to avoid condensation at the plastik cover of wells. the even
more annoying thing is that the condensation pattern changes from very fine to
bigger droplets within ca. 10 hours. while the small droplets are actually not so
bad, the bigger droplets cause severe problems for the transmission imaging
(introducing some lensing effects).

taking the lid off seems no option, because the wells may dry out over 10-24
hours.

we tried putting some oil on the cover (from the inside of course), hoping that
it would be more hydrophobic and thus prevent condensation, but
unfortunately that only helped for a few hours...

Anyone any ideas?

Thanks already!!!

Tischi (ALMF@EMBL Heidelberg)

[Romin, Yevgeniy/Sloan Kettering Institute]

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We use these mebranes that have a hydrophobic side to place on top of dishes instead of lids.  It prevents drying out and has no condensation. I haven't seen these for 24-well plate, but I'm sure they have them.  we go ours through our Zeiss applications specialist (no commercial interest)

Yevgeniy



-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Christian Tischer
Sent: Tuesday, February 21, 2012 4:43 AM
To: [hidden email]
Subject: avoid condensation??

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Hello,

we are using an inverted microscope and need to do long term transmission imaging in a 24-well dish. the whole central part of microscope is surrounded by a box keeping it at 37 degrees and providing CO2.

so far, we fail to avoid condensation at the plastik cover of wells. the even more annoying thing is that the condensation pattern changes from very fine to bigger droplets within ca. 10 hours. while the small droplets are actually not so bad, the bigger droplets cause severe problems for the transmission imaging (introducing some lensing effects).

taking the lid off seems no option, because the wells may dry out over 10-24 hours.

we tried putting some oil on the cover (from the inside of course), hoping that it would be more hydrophobic and thus prevent condensation, but unfortunately that only helped for a few hours...

Anyone any ideas?

Thanks already!!!

Tischi (ALMF@EMBL Heidelberg)


 
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[kspencer007]

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Hi Tischi;
        I've used similar setups where I put a layer of light mineral oil over the top of the media in the wells. This prevents evaporation (I've gone up to 48 hours), and the pH seemed to be stable. Some cells may not tolerate this, but it's a simple thing to try.
        Kathy Spencer
        The Scripps Research Institute



-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Christian Tischer
Sent: Tuesday, February 21, 2012 1:43 AM
To: [hidden email]
Subject: avoid condensation??

*****
To join, leave or search the confocal microscopy listserv, go to:
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Hello,

we are using an inverted microscope and need to do long term transmission
imaging in a 24-well dish. the whole central part of microscope is surrounded by
a box keeping it at 37 degrees and providing CO2.

so far, we fail to avoid condensation at the plastik cover of wells. the even
more annoying thing is that the condensation pattern changes from very fine to
bigger droplets within ca. 10 hours. while the small droplets are actually not so
bad, the bigger droplets cause severe problems for the transmission imaging
(introducing some lensing effects).

taking the lid off seems no option, because the wells may dry out over 10-24
hours.

we tried putting some oil on the cover (from the inside of course), hoping that
it would be more hydrophobic and thus prevent condensation, but
unfortunately that only helped for a few hours...

Anyone any ideas?

Thanks already!!!

Tischi (ALMF@EMBL Heidelberg)

[James Denegre]

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We cover the medium with a thin layer of mineral oil, which has been
saturated with gas; in our case CO2/O2/N2, at 37 degrees, and don't use
the lid.  Our mineral oil is from Sigma, other mineral oils may extract
ingredients from the medium. When saturated, gas exchange across the oil
is fine, as the absorption coefficients of CO2 and O2 are similar to those
in water (Kubie, JBC, 1927). We perform imaging for up to five days in
this manner, with no loss of medium and no condensation.


Jim Denegre
The Jackson Laboratory







On 2/21/12 4:42 AM, "Christian Tischer" <[hidden email]> wrote:

>*****
>To join, leave or search the confocal microscopy listserv, go to:
>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>*****
>
>Hello,
>
>we are using an inverted microscope and need to do long term transmission
>imaging in a 24-well dish. the whole central part of microscope is
>surrounded by
>a box keeping it at 37 degrees and providing CO2.
>
>so far, we fail to avoid condensation at the plastik cover of wells. the
>even
>more annoying thing is that the condensation pattern changes from very
>fine to
>bigger droplets within ca. 10 hours. while the small droplets are
>actually not so
>bad, the bigger droplets cause severe problems for the transmission
>imaging
>(introducing some lensing effects).
>
>taking the lid off seems no option, because the wells may dry out over
>10-24
>hours.
>
>we tried putting some oil on the cover (from the inside of course),
>hoping that
>it would be more hydrophobic and thus prevent condensation, but
>unfortunately that only helped for a few hours...
>
>Anyone any ideas?
>
>Thanks already!!!
>
>Tischi (ALMF@EMBL Heidelberg)

[Boswell, Carl A - (cboswell)]

Hi Tischi,
Have you considered humidifying the heating/gas chamber?  There are some very simple methods to keep humidity high in a closed environment, including running the gas through water using an aquarium aerator.  Cotton balls or crumpled paper towels in shallow dishes provide lots of moist surface area.
Good luck,
C


Carl A. Boswell
520-742-6131


-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Christian Tischer
Sent: Tuesday, February 21, 2012 2:43 AM
To: [hidden email]
Subject: avoid condensation??

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hello,

we are using an inverted microscope and need to do long term transmission imaging in a 24-well dish. the whole central part of microscope is surrounded by a box keeping it at 37 degrees and providing CO2.

so far, we fail to avoid condensation at the plastik cover of wells. the even more annoying thing is that the condensation pattern changes from very fine to bigger droplets within ca. 10 hours. while the small droplets are actually not so bad, the bigger droplets cause severe problems for the transmission imaging (introducing some lensing effects).

taking the lid off seems no option, because the wells may dry out over 10-24 hours.

we tried putting some oil on the cover (from the inside of course), hoping that it would be more hydrophobic and thus prevent condensation, but unfortunately that only helped for a few hours...

Anyone any ideas?

Thanks already!!!

Tischi (ALMF@EMBL Heidelberg)