Related Experiment Video
Updated: Aug 20, 2026
Undecalcified Bone Preparation for Histology, Histomorphometry and Fluorochrome Analysis
Published on: January 8, 2010
Differentiation of bone and soft tissues in formalin-fixed, paraffin-embedded tissue by using methylene blue/acid
Katherine A Rider1, Lisa M Flick
1Division of Biology, Alfred University, Alfred, New York 14802, USA.
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.
Study Design:
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.
Conclusion:
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.
Related Concept Videos
01:03Fixation and Sectioning
The simplest type of preparation is the wet mount, in which the specimen is placed in a drop of liquid on the slide. A liquid specimen can be directly deposited on the slide using a dropper. Solid specimens, such as skin scraping, can be placed on the slide before adding a drop of liquid to prepare the wet mount. Sometimes the liquid is simply water, but stains are often added...
01:26Differential Staining Technique