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

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 t...

Full description

Saved in:
Bibliographic Details
Published in:PloS one 2015-01, Vol.10 (1), p.e0116662-e0116662
Main Authors: Calve, Sarah, Ready, Andrew, Huppenbauer, Christopher, Main, Russell, Neu, Corey P
Format: Article
Language:English
Subjects:
Animals
Biocompatibility
Biomedical materials
Bone (cortical)
Bone and Bones - cytology
Cartilage
Cartilage - cytology
Cattle
Chondrocytes
Chondrocytes - cytology
Clearing
Confocal
Confocal microscopy
Connective tissues
Cortical bone
Embedded systems
Embedding
Fructose
Head - physiology
Hindlimb - cytology
Histological Techniques - methods
Ligaments - cytology
Light transmission
Mathematical morphology
Mice
Microscopy
Microscopy, Confocal - methods
Musculoskeletal system
Nondestructive testing
Osteocytes - cytology
Pattern formation
Refractive index
Refractivity
Solutions - chemistry
Tissues
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0116662