liquid spilling of slides - (Oct/15/2006 )

Dear all:
Iam doing some immunohistochimistry on brain slices that has been mounted on some slides -- my problem is that most of the time the liquid I put on it, ie blocking buffer, anitbody solution would spill off -- I know this because sometimes the slides become dry. Ive tried a few things already.

1) some kind of pap pen -- for some reason this does not seem to be enough -- it think one problem is that I cannot get the slide to be at an even plane
2) putting a thin parafilm over the slide

I was wondering about maybe mixing my solutions, primary, secondary, even blocking buffer in some kind of glyerol solution and cover slipping it... will this work? I am thinking maybe I would need to incubate it for longer if this was the case, but for how much longer?

thanks!
al

I would still suggest using the hydrophobic pen or the pap pen. try to make a thick marking around the sections. A thin marking might not help. Also check if u need to order a new pen, may b th eone u r using is getting old.

And try to limit the amount of liquid u r using for blocking or for antibodies.

I use the ImmEdge pen (Vector labs) and never had this problem. Scolix advice is good, and sometimes you have to push the pen on a piece of paper towel to get it flowing. If your sample is small you should be able to keep the liquid in a drop with surface tension so that it doesn't flow off the sample.

Is it possible your liquid is drying out, not coming off the slide? I use a "humidity chamber" that I made by placing paper towels in the bottom of a slide box and covering with water. Then I place the slides across the holders over the paper towels. When doing long incubations I close the chamber and for overnight I place at 4 degrees.