Kathy,

You did not say what type of bone preparation?  Paraffin or
methymethacrylate?  

There is a classic method using undecalcified sections ground and mounted
in heat melted Canada balsam or air injection method, a lot of work! 

You need to get this publication, Taylor RI, et al Variation of the Holmes
method for histologic staining of bone canaliculi, J of Histotechnology
16(4):355-357.  

But we have always been able to demonstrate canaliculi in PMMA embedded
bone, ground sections, acid etched and stained with toluidine blue at pH 8
0.1M phosphate buffer (Eurell and Sterchi) but have also seen them in same
type of preparation with other stains, Stevenels blue, MacNeals tetrachrome.  

Lacunae are much easier to see since they are much larger, and you should
see these with any preparation as the osteocyte resides there.  You can
create empty looking lacunae by over exposure to acid decalcifiers (a no no
for diagnositic work, looks like necrotic bone with empty lacunae).  The
effects of acid "overexposure" renders the nuclei basically colorless.  But
you would have to try and neutralize the acid with a base to make the
hematoxylin work, although the osteocyte might stain a bit.    

Some cross sections of rabbit femur, decalcified with endpoint, then
stained with hematoxylin longer, and a light eosin y can show these
features.  Also, to really demonstrate them, take a photomic of H&E, then
scan onto computer in black and white or gray scale!  Surprising what shows
up.  

Don't overstain with eosin, or just use a hematoxylin stain, do not
differentiate or do any acid wash, just blue the section and coverslip. 

Personally, have never had a problem seeing these structures with careful
endpoint determinations and proper staining OR acid etched ground sections
embedded in PMMA. 

 

Hello All!

I have a  researcher here who would like a thionin and picric acid stain 
for lacunae and canaliculi. I would really like to avoid the use of 
saturated picric acid. Are there any other stains that anyone knows of that 
stains the same or similar? Could you share your procedure with me? Thanks 
a million!!


Kathy Cormier
Histology Supervisor
MIT- Division of Comparative Med 

Gayle Callis
MT,HT,HTL(ASCP)
Histopathology Supervisor
Veterinary Molecular Biology - Marsh Lab
Montana State University - Bozeman
19th and Lincoln St
Bozeman MT 59717-3610

406 994-6367
406 994-4303 (FAX)