ArticlePloS one2017
Intravital imaging of osteocytes in mouse calvaria using third harmonic generation microscopy.
Article in PloS one, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed, 50 citations in OpenAlex.
- Label-free multimodal nonlinear microscopy enabled by an optical parametric generator.APL photonics · 2026Article
- Modulation of tocotrienol's bone effects by osteocytes: a perspective.Frontiers in pharmacology · 2026Article
- Intravital Microscopy of Osteocytes: A New Era in Bone Cell Biology.Current osteoporosis reports · 2025Review
- Optimized intravital three-photon imaging of intact mouse tibia links plasma cell motility to functional states.iScience · 2024Article
- Label-free multimodal imaging with simultaneous two-photon and three-photon microscopy and kernel-based nonlinear scaling denoising.Biomedical optics express · 2024Article
- Effect of out of plane orientation on polarization second harmonic generation of single collagen fibrils.Biomedical optics express · 2023Article
- Contributions of Resin Cast Etching to Visualising the Osteocyte Lacuno-Canalicular Network Architecture in Bone Biology and Tissue Engineering.Calcified tissue international · 2023Review
- Control of osteocyte dendrite formation by Sp7 and its target gene osteocrin.Nature communications · 2021Article
- Energy Metabolism of Osteocytes.Current osteoporosis reports · 2021Review
- Investigating the optical clearing effects of 50% glycerol in ex vivo human skin by harmonic generation microscopy.Scientific reports · 2021Article
- Third Harmonic Generation microscopy distinguishes malignant cell grade in human breast tissue biopsies.Scientific reports · 2020Article
- Towards a Connectomic Description of the Osteocyte Lacunocanalicular Network in Bone.Current osteoporosis reports · 2019Review
- Investigating Osteocytic Perilacunar/Canalicular Remodeling.Current osteoporosis reports · 2019Review
- Second and third harmonic generation microscopy visualizes key structural components in fresh unprocessed healthy human breast tissue.Journal of biophotonics · 2019Article
- 3-dimensional visualization of implant-tissue interface with the polyethylene glycol associated solvent system tissue clearing method.Cell proliferation · 2019Article
- A Method to Measure Myeloarchitecture of the Murine Cerebral CortexFrontiers in neuroanatomy · 2019Article
- Third harmonic generation imaging and analysis of the effect of low gravity on the lacuno-canalicular network of mouse bone.PloS one · 2019Article
- Harmonic Generation Microscopy 2.0: New Tricks Empowering Intravital Imaging for Neuroscience.Frontiers in molecular biosciences · 2019Review
- A Review of the Impact of Implant Biomaterials on Osteocytes.Journal of dental research · 2018Review
- In Vivo 3D Histomorphometry Quantifies Bone Apposition and Skeletal Progenitor Cell Differentiation.Scientific reports · 2018Article
Corrections and comments
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Authors and funding
13 authors at 4 institutions in 5 countries.
Funding
Abstract
Osteocytes are the most abundant cell in the bone, and have multiple functions including mechanosensing and regulation of bone remodeling activities. Since osteocytes are embedded in the bone matrix, their inaccessibility makes in vivo studies problematic. Therefore, a non-invasive technique with high spatial resolution is desired. The purpose of this study is to investigate the use of third harmonic generation (THG) microscopy as a noninvasive technique for high-resolution imaging of the lacunar-canalicular network (LCN) in live mice. By performing THG imaging in combination with two- and three-photon fluorescence microscopy, we show that THG signal is produced from the bone-interstitial fluid boundary of the lacuna, while the interstitial fluid-osteocyte cell boundary shows a weaker THG signal. Canaliculi are also readily visualized by THG imaging, with canaliculi oriented at small angles relative to the optical axis exhibiting stronger signal intensity compared to those oriented perpendicular to the optical axis (parallel to the image plane). By measuring forward- versus epi-detected THG signals in thinned versus thick bone samples ex vivo, we found that the epi-collected THG from the LCN of intact bone contains a superposition of backward-directed and backscattered forward-THG. As an example of a biological application, THG was used as a label-free imaging technique to study structural variations in the LCN of live mice deficient in both histone deacetylase 4 and 5 (HDAC4, HDAC5). Three-dimensional analyses were performed and revealed statistically significant differences between the HDAC4/5 double knockout and wild type mice in the number of osteocytes per volume and the number of canaliculi per lacunar surface area. These changes in osteocyte density and dendritic projections occurred without differences in lacunar size. This study demonstrates that THG microscopy imaging of the LCN in live mice enables quantitative analysis of osteocytes in animal models without the use of dyes or physical sectioning.
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Registered trials
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