Differentiation of bone and soft tissues in formalin-fixed, paraffin-embedded tissue by using methylene blue/acid fuchsin stain.
OBJECTIVE
To find a staining method for formalin-fixed, paraffin-embedded tissue that would distinguish bone from surrounding soft tissues, including muscle, periosteal tissue and bone marrow.


METHODS
A variety of stains were tested and compared with hematoxylin-eosin. The potential value of any given stain was evaluated based on its ability to stain bone and soft tissues different colors or shades that could be readily identified in photomicrographs. Stains were evaluated using both endochondral (tibia) and intramembranous bone (calvaria) samples.


RESULTS
In contrast to standard hematoxylin-eosin stain, which stains both bone and soft tissues pink, the methylene blue/acid fuchsin stain demonstrates remarkable contrast between bone and other tissues. Methylene blue/acid fuchsin stained bone bright pink and the surrounding soft tissues blue-purple.


CONCLUSIONS
In addition to the superior staining properties of methylene blue/acid fuchsin, other benefits of this stain include its stability, ease of use and low cost. This stain has many potential applications in the study of erosive bone disease in humans and also in animal models for research.
Rider
Katherine A
KA
Division of Biology, Alfred University, Alfred, New York 14802, USA.
Flick
Lisa M
LM
D000818:Animals; D001557:Benzenesulfonates; D001854:Bone Marrow Cells; D001842:Bone and Bones / Q000166:cytology*; D005557:Formaldehyde; D008751:Methylene Blue; D051379:Mice; D008810:Mice, Inbred C57BL; D018482:Muscle, Skeletal / Q000166:cytology; D010537:Peritoneum / Q000166:cytology; D010784:Photomicrography / Q000592:standards; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods / Q000592:standards*; D016707:Tissue Fixation
D001557:Benzenesulfonates; D005557:Formaldehyde; C086337:acid fuchsin; D008751:Methylene Blue