1. Bone CLARITY: Clearing, imaging, and computational analysis of osteoprogenitors within intact bone marrow.
- Author
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Greenbaum A, Chan KY, Dobreva T, Brown D, Balani DH, Boyce R, Kronenberg HM, McBride HJ, and Gradinaru V
- Subjects
- Animals, Cell Differentiation physiology, Cells, Cultured, Mice, Mice, Transgenic, Osteoblasts cytology, Osteogenesis physiology, SOX9 Transcription Factor metabolism, Stem Cells metabolism, Bone Marrow Cells cytology, Stem Cells cytology
- Abstract
Bone tissue harbors unique and essential physiological processes, such as hematopoiesis, bone growth, and bone remodeling. To enable visualization of these processes at the cellular level in an intact environment, we developed "Bone CLARITY," a bone tissue clearing method. We used Bone CLARITY and a custom-built light-sheet fluorescence microscope to detect the endogenous fluorescence of Sox9-tdTomato
+ osteoprogenitor cells in the tibia, femur, and vertebral column of adult transgenic mice. To obtain a complete distribution map of these osteoprogenitor cells, we developed a computational pipeline that semiautomatically detects individual Sox9-tdTomato+ cells in their native three-dimensional environment. Our computational method counted all labeled osteoprogenitor cells without relying on sampling techniques and displayed increased precision when compared with traditional stereology techniques for estimating the total number of these rare cells. We demonstrate the value of the clearing-imaging pipeline by quantifying changes in the population of Sox9-tdTomato-labeled osteoprogenitor cells after sclerostin antibody treatment. Bone tissue clearing is able to provide fast and comprehensive visualization of biological processes in intact bone tissue., (Copyright © 2017, American Association for the Advancement of Science.)- Published
- 2017
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