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PLoS One. 2015 Jan 12;10(1):e0116662. doi: 10.1371/journal.pone.0116662. eCollection 2015.

Optical clearing in dense connective tissues to visualize cellular connectivity in situ.

Author information

1
Weldon School of Biomedical Engineering, Purdue University, West Lafayette, Indiana, United States of America.
2
W. Nuhsbaum, Inc., McHenry, Illinois, United States of America.
3
Weldon School of Biomedical Engineering, Purdue University, West Lafayette, Indiana, United States of America; Department of Basic Medical Sciences, Purdue University, West Lafayette, Indiana, United States of America.

Abstract

Visualizing the three-dimensional morphology and spatial patterning of cells embedded deep within dense connective tissues of the musculoskeletal system has been possible only by utilizing destructive techniques. Here we utilize fructose-based clearing solutions to image cell connectivity and deep tissue-scale patterning in situ by standard confocal microscopy. Optical clearing takes advantage of refractive index matching of tissue and the embedding medium to visualize light transmission through a broad range of bovine and whole mount murine tissues, including cartilage, bone, and ligament, of the head and hindlimb. Using non-destructive methods, we show for the first time intercellular chondrocyte connections throughout the bulk of cartilage, and we reveal in situ patterns of osteocyte processes and the lacunar-canalicular system deep within mineralized cortical bone. Optical clearing of connective tissues is expected to find broad application for the study of cell responses in normal physiology and disease pathology.

PMID:
25581165
PMCID:
PMC4291226
DOI:
10.1371/journal.pone.0116662
[Indexed for MEDLINE]
Free PMC Article

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