How do I incubate slide-mounted sections with Ab?
e.pietrzak at mailbox.uq.edu.au
Thu Oct 17 20:14:07 EST 1996
In article <Pine.SGI.3.94.961017130003.19844A-100000 at leonardo.ls.huji.ac.il> broide at LEONARDO.LS.HUJI.AC.IL (Ron BROIDE) writes:
>To all who read this,
>I have a technical question in immunohistichemistry:
>I have started doing some immunohistochemistry on slide-mounted frozen
>sections in a lab that uses parafilm to cover slides for the purpous of
>incubating with labeled riboprobes. This method works quite well for
>them, but doesn't seem to work for my antibody incubations. Instead, what
>I end up with is a gradient of labeling from the top to the bottom of the
>I've seen people surround their slide-mounted sections with a ring of nail
>polish or grease in order to form a well for their antibody incubations.
>This seems a bit messy. I've also seen some plastic/rubber chambers that
>can stick onto glass slides and form a small incubation chamber (not the
>ones from Labtech for cell culture). This looked like a neat trick.
>1) Does anyone know what the latest technique is for antibody incubations
>on slide-mounted sections?
>2) Does anyone know who sells these type of plastic/rubber chambers that
>can stick onto glass slides?
>Thanks for the help,
>Ron S. Broide, Ph.D.
>Department of Biological Chemistry
>Institute of Life Science
>Tel: 972-2-658 5450
>FAX: 972-2-652 0258
>broide at leonardo.ls.huji.ac.il
You can use special DAKO pen to outline the shape of your section.
It forms a well around it.
Even simpler method is to put a drop on your section without anything.
The slide is much more hydrophobic than your section, the drop will not run.
The magic of succes is:
1/ to keep it in a wet chamber so it does not dry,
2/ to handle it gently, without jerking, so it does not run off,
3/ and to wipe the slide dry around the section after washing.
The gradient you had was probably due to drying up.
Eva Pietrzak, PhD
Dept. of Dentistry
University of Queensland
More information about the Immuno