RE: immuno/floating tissue
1)Try BD Gold Seal Ultrastick (TM) slides or Polysciences SectionLock (TM)
2)Let the water from the flotation bath drain off the slide 3-5 minutes (by
standing the slide up against the outside edge of the float bath or in a
slide draining rack) before placing the slide flat on the slide warming
3)Leave the slide on the very warm slide warming table (approximately 5
degrees less than the meltpoint of the paraffin you are using)until the
paraffin is completely melted and the section glistens.
NOTE:If the slide has not been properly drained before placement on the
warming table small (often undetectable to the naked eye) bubbles will form
between the section and the slide. Thus decreasing any adherance you may
think you are gaining by using costly slides and expensive additives.
My two cents learned the hard way doing hard tissues, Good luck
Donna Montague, M.S.
Physiology/Biophysics and Orthopaedic Surgery
University of Arkansas for Medical Sciences
From: Vicki Kalscheur [mailto:email@example.com]
Sent: Friday, July 20, 2001 8:56 AM
Subject: immuno/floating tissue
I am a graduate student working on my MS in the Comparative Orthopedic
Research Lab at the UW-Madison.
My project involves examining chondrocyte viability in bovine patella
samples over time. Part of my project is examining the articular
cartilage for apoptotic chondrocytes. I will be using Anti-Caspace 3,
Anti-PARP, and Tunel assays to do this.
I am, however, having a problem regarding this procedure. It seems
that my samples will not adhere to the slides during the assays.
After about 4 washes (only 1/4 of the way through each assay) the
sections fall off the slides and float in the solution.
I have tried Aminoalkylsilane coated slides, and a product called HALT
to help adhere the sections but have had no luck. I am currently
using EDTA to decalcify the sections and frozen sectioning. I have
considered using paraffin embedded samples to secure my samples more
effectively but am afraid of the damage to the chondrocytes during
heating. We also plan on trying some free-floating analysis.
I would greatly appreciate any suggestions anyone could provide. My
email is firstname.lastname@example.org. My fax is 608-263-7930 (please put
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