Hi everybody,

I am fairly new to all Histotechnology processes. We mainly work in our lab 
with mineralized bone with implants, so we fix them, dehydrate them, embed them 
in MMA, cut them, glued them to slides, ground them manually and then stain 
them. I need help in some parts of the process that need improvement:


  1.  We use Loctite 4471 on the samples, put them under vacuum, apply them to 
the slide and more vacuum. There's good results but there are some bubbles that 
still appear. Suggestions?
  2.  I can't seem to get the yellow/orange on bone with the Van Gieson stain. 
I've been doinga preheat at 55 °C, etching, rinse in DI water, Sanderson´s 
Rapid Bone Stain, running tap water, Van Gieson (commercial from DHM) for 30 
seconds to 5 minutes, paper dry and quickly dehydrate with 100% EtOH. What am I 
doing wrong?
  3.  The stain's been running when I add the glue to cover slip the slides. 
Especially the green color. Why does this happen?
  4.  Cover slipping is done with cyano acrylate, under suction as before, but 
bubbles still appear. I've tried placing the cover slip on an angle and wait 
for the glue to evenly spread, but the bone has just too many pores and air 
bubbles appear. Please help.

I greatly appreciate any help on the subject.

Best,

Rodolfo De la Vega, MD.
Research Fellow
Laboratory for Musculoskeletal Research and Innovation
Massachusetts General Hospital - Jackson Building 1120



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