The use of Commercially Available Adhesive Tapes to Preserve Cartilage and Bone Tissue Integrity During Cryosectioning

Abstract

The use of fluorescent tags to monitor protein expression and to lineage-trace cells has become a standard complement to standard histological techniques in the fields of embryology, pathology and regenerative medicine. Unfortunately, traditional paraffin embedding protocols can substantially diminish or abolish the native emission signal of the fluorophore of interest. To preserve the fluorescent signal, an alternative is to use cryosectioning; however, this can often result in undesirable artefacts such as tearing or shattering – particularly for mineralized tissues such as bone and cartilage. Here we present a method of using a commercially available tape to stabilize murine femur tissue, thus allowing for cryosectioning of cartilage and bone tissues carrying fluorescent tags without the need for demineralization.

Keywords: :

• bone
• cartilage
• chondrocytes
• cryo-sectioning
• demineralization
• fluorescent imaging
• growth plate
• histology
• mouse
• osteoblasts
• transgenic

Author contributions

FVM and MAS conceived of the project. MAS and DDP carried out the experiments with help from JWH. FVM, MAS and DDP wrote the manuscript.

Acknowledgments

We gratefully thank Ashlie Muñoz for technical assistance.

Financial competing interests disclosure

This work was supported by NIH NIAMS AR064462 and AR069700 and by the USC STAR Program (to DP). This paper is subject to the NIH Public Access Policy. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

The use of animals in this research was approved by the University of Southern California’s Animal Use Committee.

Additional information

Funding

This work was supported by NIH NIAMS AR064462 and AR069700 and by the USC STAR Program (to DP). This paper is subject to the NIH Public Access Policy. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed