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Bone. 2022 Jan;154:116257. doi: 10.1016/j.bone.2021.116257. Epub 2021 Nov 13.

NAD(P)H autofluorescence lifetime imaging enables single cell analyses of cellular metabolism of osteoblasts in vitro and in vivo via two-photon microscopy.

Bone

Kevin Schilling, Edward Brown, Xinping Zhang

Affiliations

  1. Center for Musculoskeletal Research, University of Rochester, School of Medicine and Dentistry, Rochester, NY 14642, USA; Department of Biomedical Engineering, University of Rochester, Rochester, NY 14642, USA.
  2. Department of Biomedical Engineering, University of Rochester, Rochester, NY 14642, USA.
  3. Center for Musculoskeletal Research, University of Rochester, School of Medicine and Dentistry, Rochester, NY 14642, USA; Department of Biomedical Engineering, University of Rochester, Rochester, NY 14642, USA. Electronic address: [email protected].

PMID: 34781049 PMCID: PMC8671374 DOI: 10.1016/j.bone.2021.116257

Abstract

Two-photon fluorescence lifetime microscopy (2P-FLIM) is a non-invasive optical technique that can obtain cellular metabolism information based on the intrinsic autofluorescence lifetimes of free and enzyme-bound NAD(P)H, which reflect the metabolic state of single cells within the native microenvironment of the living tissue. NAD(P)H 2P-FLIM was initially performed in bone marrow stromal cell (BMSC) cultures established from Col (I) 2.3GFP or OSX-mCherry mouse models, in which osteoblastic lineage cells were labelled with green or red fluorescence protein, respectively. Measurement of the mean NAD(P)H lifetime, τ

Copyright © 2021 Elsevier Inc. All rights reserved.

Keywords: 2-photon microscopy; 2P-FLIM; 2P-PLIM; Bone repair; Energy metabolism; Glycolysis; Intravital imaging; Osteoblasts; Oxidative phosphorylation

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