[Histonet] Bone processing help

[De La Vega Amador, Rodolfo Enrique]

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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