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Shalin Mehta <[log in to unmask]> writes:

> After a month or so, I will need to use upright and inverted confocals with
> different capabilities located 1 hour bus journey apart. 
> I have been searching for culture vessel that allows live cell microscopy with
> following features (decreasing order of preference).
> 1. Can be used with both inverted and upright microscopes for live cell
> imaging
> 2. multiple chambers for multiplexing measurements
> 3. ease of transportation.

Our solution to this type of situation is to make our own sealed
chambers. You take a standard 1mm slide and drill 1 or several holes
in it. Glue a coverslip to one side. Now you have 1 or several round
holes with a coverslip on one side. You grow your cells on small
coverslips fill the holes with media and invert the cells onto the
chamber and seal with wax. You need to have media that is CO2
independent, but most cells seem to survive in these conditions.

The resulting chambers are very stiff and robust. By filling the wells
with media the cells are well protected from mechanical shocks.
Optically both side of the chamber are a standard coverslip, with only
1mm of liquid between them so should work for most setups. 

To reuse you simply scrape of the wax, throw away your coverslip with
cells on it and acid wash the chamber. The only problem is drilling
the glass is a pain. Be very careful as glass dust is nasty stuff.

Ian



>  
> I tried samples from this:
> 1. Nunc -- Lab-tek chambered slides - For fixed samples, it works nicely but
> for live cells the thick bottom of slide won't work with objectives on
> inverted microscope. It cannot be turned upside down to view adherent cells
> under upright microscope. Not easy to transport.
> 2. ibidi -- u-slide 6 - Works with inverted scope because bottom is
> coverslip-like. Can be turned upside down (for using with upright) when wells
> of all channels are sealed. But then condenser cannot come close enough
> because slide is resting on wells and hence cannot get good illumination
> required for taking reference images. Relatively easy to transport live cells.
> 3. Nunc -- Opticell - (Yet to try) appears to work with both upright and
> inverted scope. Comes with special transportation envelope. Its memberanes are
> 75 micron thick (less than half the coverslip thickness), so I am concerned
> about errors due to wrong coverslip thickness. Lacks multiple chambers and
> requires large volume of reagents.
>  
> Please share your suggestions on modifying one of this or using another
> product?
>
> Regards
> Thanks
> Shalin
>
> --
> My co-ordinates:
> Shalin Mehta, Graduate student
> Graduate Programme in Bioengineering, NUS, Singapore
> Email: shalin {dot} mehta {at} gmail {dot} com
> Blog: electricsbm.blogspot.com
> Mobile: +65 90694182