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We have used an intermediate solution: we detach the coverslips from the used glass bottom petri dishes (e.g. mattek). Then we clean the dishes without fear. After that we glue back coverslips of proper thickness (#1.5) in the bottom using some easy to remove glue (Sylgard works OK, it needs to cure completely and cannot be used in excess as it may leak and cover some of the glass from the inside). Next we rinse and sterilize the plates using ethanol and UV exposure. Finally, we culture-treat just the glass portion of the refurbished dish if needed.. 
I said intermediate solution because still wastes a lot of time... But sometimes we have more time than money. The part we hate the most is the fearless washing. 

-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of Karl Garsha
Sent: Wednesday, December 15, 2004 9:12 AM
To: [log in to unmask]
Subject: Re: glass bottom cell culture plates: cleaning / reuse for cell culture?


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Hello Guenter,
I still use old fashioned reusable glassware for cell culture. The chambered coverslips and glass-bottom multiwell plates are relatively fragile, and they are diffucult to clean without breaking. The glue that holds the glass to the plastic can disintegrate and you end up with leaky chambers (which is very frustrating and messy). The surface treatment necessary depends on the cell lines being cultured, but I would imagine that the wells could be re-treated without too much of a problem. The hard parts are washing to the point of scrupulous cleanliness and sterilizing. One approach would be to wash the culture vessels with soap/water, rinse well, soak in 70% ethanol and let dry under a bacteriocidal UV lamp in a laminar flow hood. Then coat with whatever substrate is necessary for cell adherence under sterile conditions. It is possible that some bacterial endotoxins could left behind which would complicate immunological assays, so the washing would be critical. The glues on the commercial chambered coverglasses will not hold up long under this sort of regimen; I'm pretty sure these things are thoughtfully engineered to fall apart and you might get quite a few leakers. Cutting out the ethanol soak might help in this respect. Making your own reusable chambered coverglasses would probably be more successful also. -Karl

Guenter Giese wrote:

> Search the CONFOCAL archive at 
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Dear microscopists,
>
> multiwell glass bottom cell culture plates suitable for confocal 
> microscopy are quite expensive (15 to 50 € / $ each), and we would 
> like to reuse them if possible. We do time lapse microscopy 
> experiments on living cells
>
> Reuse of glassware in cell culture was (is still?) commonplace, but 
> the combination of glass and plastic and the high surface to volume 
> ratio of multiwell plates may be disadvantageous in this respect.
>
>
> Two scenarios:
>
> i) Only some wells of the multiwell plate are used for cell culture in 
> one experiment: Used wells could be treated individually after the 
> experiment to kill any living cells / bacteria and to inactivate 
> viruses.
> - Which chemicals / methods are safe in preventing collateral damage
> to neighboring, unused wells?
>
> ii) All wells are used:
> - By which method could one clean and treat the plates to allow proper 
> growth of the cells (cleaning / surface treatment protocol)?
>
> I know that it is wise to use new material, but at least for 
> preliminary checks we would like to reuse the plates. One could also 
> preclean new plates to allow comparison of results (after comparison 
> of results obtained with used vs unused plates).
>
> Guenter
>
> ------------------------------------------
> Dr. Guenter Giese
> Light Microscopy Facility
> Dept. of Biomedical Optics, MPI fuer Medizinische Forschung Jahnstr. 
> 29, D-69120 Heidelberg, Germany Phone (+49) 6221-486-360 (Fax: -325)
> e-mail: [log in to unmask]
> http://www.mpimf-heidelberg.mpg.de/~ggiese/lightmicro


--
Karl Garsha
Light Microscopy Specialist
Imaging Technology Group
Beckman Institute for Advanced Science and Technology
University of Illinois at Urbana-Champaign
405 North Mathews Avenue
Urbana, IL 61801
Office: B650J
Phone: 217.244.6292
Fax: 217.244.6219
Mobile: 217.390.1874
www.itg.uiuc.edu