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Julia -

Here's a method that may work for you. It is a well known botanical 
method that I have used many times without any toxicity problems. It's 
cheap and easy, and it seals well. It's autoclavable, and it keeps 
forever un-refrigerated. I have some always on hand. The sealant is 
called VaLaP and it's cheap and non-toxic. Get yourself some plain 
Vaseline (unscented etc.) and some ordinary canning paraffin from the 
supermarket, and some lanolin, which you can get in bulk from VWR or 
Fisher where they list it as "Wool Fat". Mix them well 1:1:1 (equal 
parts) at 60C. I have a little hotplate that I keep just for my VaLaP, 
always ready. When you need it melt it at 60C and apply it with a Q-tip. 
I dab it on the corners of the coverslip just to keep it in place, and 
then paint around the edge to seal. It is very sticky so it seals well 
and will stick to polycarbonate or glass, and yet it is not difficult to 
handle once you have tried it a couple of times.

I have kept fern material alive for an extended period for a week or 
more of continuous time-lapse. I autoclave in a little beaker. No 
contamination or leakage. Obviously, you don't want to get in on the 
objective, but If you get it on the lens it cleans off well with "Sparkle".

Try it. you may find it is just what you need.

Regards,

Phil Lintilhac

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On 4/22/24 1:43 PM, Julia Edgar wrote:
> *****
> To join or leave the confocal microscopy listserv or to change your email address, go to:
> https://lists.umn.edu/cgi-bin/wa?SUBED1=confocalmicroscopy&A=1
> Post images onhttp://www.imgur.com  and include the link in your posting.
> *****
>
> Dear Confocal Microscopy List
> Years ago I got a very useful protocol from somebody on the List for making live imaging dishes. Basically, one burred holes in a Petri dish and glued a suitable coverslip beneath using World Precision Instruments, #SYLG184. We continue to use Slyfard (R) 184, but we can't keep the cells alive. It seems like the glue is somehow toxic to the cells - yet we had great success in the past.
>
> Any advice will be greatly appreciated.
> Best wishes
> Julia
>