Immuno on freezing microtome sections.

From:Neuropathology <> (by way of Sue Danielson <>)


Our lab is just beginning to do this very thing.  We first stained thinner
sections (15-20 microns) with the PGP ( using a fluorescent tag) and looked
at them using a regular microscope (an olympus BX40).

Now that our immuno procedure has been optimized,  we are cutting a series
of thicker sections (from 50-100 microns), staining them as free floating
sections for PGP, collagen IV and Ulex and attempting to look at them with
confocal microscopy.  From what I have read, immunofluorescence staining on
free-floating sections is the way to go.

Please contact us directly if we can assist you in any way.

Susan Danielson MS
Neuromuscular Lab Coordinator
Dept. Neurology, Medical College of Wisconsin
ph:  414.805.3836
fax:  414.454.7905

I 've been asked to cut 50 micron freezing microtome sections of a skin
biopsy and immunostain with PGP 9.5. I'm happy (??!!) to cut the sections
but would welcome any advice on the immunostaining. Is it better to free
float the sections through the reagents or mount the sections onto slides,
air dry and then stain? Is immunofluorescense best or can I use the standard
immuno method as used for cryostat sections?

Bob Quilty
Dept of Neuropathology
Frenchay Hospital

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