immuno/floating tissue

From:Vicki Kalscheur <>

     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  My fax is 608-263-7930 (please put 
     attention Beth at the top)

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