LabTek chambered coverglass vs. chamber slide

[Rachael Lim]

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Hi all,

Would like to ask anyone who has used LabTek II's Chambered Coverglass and
Chamber Slide.

I read from the FAQ on their website, that the Chamber Slide's upper
structure is made of polystyrene, which is not compatible with many fixation
reagents; and thus, it is designed for upper structure and gasket removal
before fixing, staining and coverslipping.

However, i found from their comparison chart that the upper structure of
their Chambered Coverglass is also made of polystyrene, BUT the upper
structure is non-removable. Why is it so? Does this mean that fluorescence
staining has to be done elsewhere prior to transfer into the Chambered
Coverglass? And are the two products suitable for fluorescence work with
inverted microscope? I have the impression that Chambered Coverglass is
great for inverted microscope, and Chamber Slide is for upright microscope.

Please advise. Thank you!

Cheers,
Rachael Lim

[Morrison, Ian E]

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Rachael, the LabTek Chambered Coverglass is designed for work on an inverted 'scope; you work with a solution in the chamber which can keep the cells alive, and you can change the media (carefully!) during the your observations if you want to.  The upper structure cannot be removed as this would break the thin glass.

The Chamber Slide is built on normal thickness glass, which is strong enough to allow the polystyrene structure to be removed, and then a #1.5 coverslip can be placed over the cells (and a drop of medium) using nail varnish or vacuum grease around the edges to seal it.  The slide can then be observed on an upright or inverted 'scope, but the cells will only live for a short time.  When you remove the polystyrene structure, it is important to check that all of the silicone gasket material comes away, as a piece of this can prevent the coverslip from sitting down properly, so you may not be able to focus on the cells.  

You normally stain and fix before removing the plastic structure, but you have to use a fixative that is compatible with these materials.  Ian

--------------------------Dr. I.E.G.Morrison       {[hidden email]}------------------
                          Dept.Biological Sciences, University of Essex
                          Wivenhoe Park, Colchester, Essex CO4 3SQ
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-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]]On
Behalf Of Rachael Lim
Sent: Wednesday, October 31, 2007 5:15 AM
To: [hidden email]
Subject: LabTek chambered coverglass vs. chamber slide


Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Hi all,

Would like to ask anyone who has used LabTek II's Chambered Coverglass and
Chamber Slide.

I read from the FAQ on their website, that the Chamber Slide's upper
structure is made of polystyrene, which is not compatible with many fixation
reagents; and thus, it is designed for upper structure and gasket removal
before fixing, staining and coverslipping.

However, i found from their comparison chart that the upper structure of
their Chambered Coverglass is also made of polystyrene, BUT the upper
structure is non-removable. Why is it so? Does this mean that fluorescence
staining has to be done elsewhere prior to transfer into the Chambered
Coverglass? And are the two products suitable for fluorescence work with
inverted microscope? I have the impression that Chambered Coverglass is
great for inverted microscope, and Chamber Slide is for upright microscope.

Please advise. Thank you!

Cheers,
Rachael Lim

[Engstrom, Lars]

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Yes. The chambered coverglass is great for inverted scopes. I have used
the chambered coverglass a lot and have had no problems fixing with 10%
buffered formalin, my fixative of choice. You can probably get by with
most fixatives if you use a small volume and you shouldn't need to fix
cultured cells for very long. The only problems I have had with our set
up is that the end chambers are sometimes hard to image with our
coverglass stage insert and the plastic tab has to be cut off (a change
in manufacturing happened about 2 years ago, not sure why, anyone?). I
usually only use the center 4 wells of an 8 chambered coverglass.

The chambered slides can also be used on an inverted scope but I don't
like the small gap that remains even after removing the gasket and
therefore almost always image in the chambered cover glass with a small
volume of PBS or HBSS and a water immersion objective for higher
magnifications.

Good luck.
-Lars

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On
Behalf Of Rachael Lim
Sent: Tuesday, October 30, 2007 10:15 PM
To: [hidden email]
Subject: LabTek chambered coverglass vs. chamber slide

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Hi all,

Would like to ask anyone who has used LabTek II's Chambered Coverglass
and
Chamber Slide.

I read from the FAQ on their website, that the Chamber Slide's upper
structure is made of polystyrene, which is not compatible with many
fixation
reagents; and thus, it is designed for upper structure and gasket
removal
before fixing, staining and coverslipping.

However, i found from their comparison chart that the upper structure of
their Chambered Coverglass is also made of polystyrene, BUT the upper
structure is non-removable. Why is it so? Does this mean that
fluorescence
staining has to be done elsewhere prior to transfer into the Chambered
Coverglass? And are the two products suitable for fluorescence work with
inverted microscope? I have the impression that Chambered Coverglass is
great for inverted microscope, and Chamber Slide is for upright
microscope.

Please advise. Thank you!

Cheers,
Rachael Lim