JB4 section loss and staining

From:Gayle Callis

You can use Plus charge slides, but the problem lies more in going through
solvents during staining. 

Gill 3 is excellent for GMA sections. You can never get the same depth of
staining with 2um GMA sections since there isn't enough tissue there to
stain and be dark enough. Same with very thin paraffin sections.  

We air dried sections, DO NOT REHYDRATE THEM, immersed thim directly into
Gill 3 for 10 min, rinsed three washes with distilled water 10 dips each,
we never used acetic acid rinse, blued in Scotts Tap water, and rinsed in
tap water very briefly. Sections were air dried, and then immersed directly
into Eosin y, preferred is eosin phloxine, rinse VERY FAST through 2
changes of 95% alcohol - air dry sections again and coverslip DRY sections
with mounting media.  Prolonged soaking in xylene creates a parched earth
effect in open tissue spaces, this is the plastic - ugly artifact.   

Extended rinsing in alcohol releases GMA from slide and extensive rinsing
causes this plastic to visibly expand from slide surface. Air drying does
not hurt tissues in GMA, since the plastic cannot be removed anyway.  
Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology - Marsh Lab
Montana State University - Bozeman
S. 19th and Lincoln St
Bozeman MT 59717-3610

406 994-6367 (lab with voice mail)
406 994-4303 (FAX)

email: gcallis@montana.edu

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