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Also, are you re-using the dishes? The age and use of your dish could also
be impacting your cells. Overly UV-ing could also be breaking down the
plastic dish as well.

Much regards,
Quinn

On Mon, Apr 22, 2024 at 1:25 PM Quinn Lee <[log in to unmask]> wrote:

> Hello Julia,
>
> Is the Sylgard elastomer old? Or possibly not curing properly? I also use
> Sylgard 184 to attach new cover slips to MatTek Glass Bottom Dishes, which
> seems similar to your setup. Mine is from Dow Chemical (NC9285739) and I
> have had no issue so far. Usually when my cells die it is usually from
> harsh transfection, insufficient recovery during knockdown and
> transfection, and/or the coverslip was insufficiently coated with
> attachment factors  (ie collagen).
>
> My work involves making biomimetic surfaces, thus I cannot use vendor-made
> dishes as-is. If you similarly have your own reasons as to not use
> vendor-made dishes, you possibly try to use them as a possible control as
> you troubleshoot?
>
> Much regards,
> Quinn
>
> On Mon, Apr 22, 2024 at 1:21 PM Christopher King <[log in to unmask]>
> 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 on http://www.imgur.com and include the link in your posting.
>> *****
>>
>> Hi Julia,
>>
>> Considering the cost of the time to make the dishes and find a suitable
>> glue, it might be worth considering to purchase glass bottom dishes
>> online.
>> There are a number of companies-you can even get them precoated with
>> things
>> like polyLysine, matrigel, collagen, etc.
>>
>> Best,
>> Chris
>>
>> On Mon, Apr 22, 2024, 10:44 AM Julia Edgar <[log in to unmask]>
>> 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 on http://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
>> >
>> >
>>
>