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J Biomed Opt. 2016 Apr 30;21(4):46005. doi: 10.1117/1.JBO.21.4.046005.

Label-free identification of macrophage phenotype by fluorescence lifetime imaging microscopy.

Author information

1
University of California Irvine, Department of Biomedical Engineering, 2412 Engineering Hall, Irvine, California 92697, United States.
2
University of California Irvine, Department of Pharmacological Sciences, 2412 Engineering Hall, Irvine, California 92697, United States.
3
University of California Irvine, Department of Chemistry, 2412 Engineering Hall, Irvine, California 92697, United States.

Abstract

Macrophages adopt a variety of phenotypes that are a reflection of the many functions they perform as part of the immune system. In particular, metabolism is a phenotypic trait that differs between classically activated, proinflammatory macrophages, and alternatively activated, prohealing macrophages. Inflammatory macrophages have a metabolism based on glycolysis while alternatively activated macrophages generally rely on oxidative phosphorylation to generate chemical energy. We employ this shift in metabolism as an endogenous marker to identify the phenotype of individual macrophages via live-cell fluorescence lifetime imaging microscopy (FLIM). We demonstrate that polarized macrophages can be readily discriminated with the aid of a phasor approach to FLIM, which provides a fast and model-free method for analyzing fluorescence lifetime images.

PMID:
27086689
PMCID:
PMC4833856
DOI:
10.1117/1.JBO.21.4.046005
[Indexed for MEDLINE]
Free PMC Article

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