How do I incubate slide-mounted sections with Ab?

Eva Pietrzak 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
>slide.

>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 Broide

>Ron S. Broide, Ph.D.
>Department of Biological Chemistry
>Institute of Life Science
>Hebrew University
>Jerusalem, 91904
>Israel
>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



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