my tissue sections are very fragile ! - (Jul/24/2008 )
I'm using frozen sections (10um thick) for my IHC. I fix the sections in 4% formaldehyde (15 min) before blocking them. however I've found them very fragile; when I'm working with lens x40 and it touches the slide it makes sections fall apart ... !!! sections are perfect with H&E staining though ... Has any one experienced the same problem ever? what do you think I can do to make my sections stronger ?
of course they're fragile they are 10 um thick
i take it you are using coverslips, vectorsheild (or the like) and nail varnish
and the tissue is held in place by apes coated slides
and you do press the tissue hard onto the slide when you section
what kind of tissue is it?
Well, tissues are from an insect (locust) ganglion ... I haven't been using any mounting medium so far, I just put one or two drops of pbs and then put the coverslip on the top of it, I seal them with nail polish then. Do you think using a mounting medium (like permount ) makes any difference?
Mounting media helps to preserve the staining. In case of fluorescence staining, helps preserve the fluorescence. Without mounting media, one has to analyze the slides immediately.
Yes, but I mean permanent mounting medium; Since it gets hard after a while, I thought it may help to make the sections more robust before I study them under the microscope?
i seem to remember vector does one
so it does