Cover slip for confocol?? - (Aug/25/2005 )

Hi,
I have been using the labtek 4 chamber glass slides for my confocal. Now I have to do similar experiments however, I nned to fix the cells with acetone. The labtek chamber slides cannot be used with acetone as they corrode.

I was wondering if anyone has been growing cells on coverslips and then fixing and all. If so can he/she pass me the catalog number for the coverslips and also the company name (US only). I was thinking of placing these coverslips in a 24 well plate and then seeding the cells. Is this the way to do it??
thnx
p..

FYI, you can buy chamber slides like Lab-tek slides that are made from glass (called Lab-tek II slides or BD Falcon Culture Slides). You can remove the top cover and fix in acetone. Some have a teflon coated slide with chambers of all sizes. I use them all the time and they are good.

Also Erie Scientific sells just the teflon coated slides without chambers which can also be used for culturing with good success (called Cel-line slides, they may need to be autoclaved for sterility first). Erie stuff can be bought from VWR also.

Here are some links:

http://www.vwrsp.com/catalog/product/index...594&resultNum=0
http://www.vwrsp.com/catalog/product/index...97&resultNum=10
http://www.cel-line.com/

ok the thing is after I fix with acetone...i want to be able to add the primary and secondary etc.
The wells bascially help me in add ing say 300 ul or so to each well. If i take off the wells I might need to incubate the whole slide in the primary solution and probably use more of the antibody mixture....
else
how do u do the primary and secondary addition after you take off the well walls??
p..

The wells are divided by a blue teflon coat which is hydrophobic. In the 8-well format I use about 25-50 ul per well for the antibody incubations. Once you remove the chamber if you find that the wells are not hydrophobic *enough* you can pap pen between wells for an added hydrophobic barrier. Works well!

Also if you prefer, you can culture cells directly on the Cel-line slides in a tiny volume, just do it in a sterile humidified larger chamber. One you fix the incubations are done with small volumes just like the cultures. There is no chamber involved with these ones.

Good luck!
Andrea

"pap pen" sorry but you lost me
p..

A pap-pen is a hydrophobic pen that marks between the wells or even between two sections on one slide so that you can work with several antibodies or conditions on one slide. See Here for more info on pap pens.

"pap pen" sorry but you lost me 
p..