Intravital microscopy of biosensor activities and intrinsic metabolic states

Seth Winfree, Takashi Hato, Richard N. Day

Research output: Contribution to journalReview article

4 Scopus citations

Abstract

Intravital microscopy (IVM) is an imaging tool that is capable of detecting subcellular signaling or metabolic events as they occur in tissues in the living animal. Imaging in highly scattering biological tissues, however, is challenging because of the attenuation of signal in images acquired at increasing depths. Depth-dependent signal attenuation is the major impediment to IVM, limiting the depth from which significant data can be obtained. Therefore, making quantitative measurements by IVM requires methods that use internal calibration, or alternatively, a completely different way of evaluating the signals. Here, we describe how ratiometric imaging of genetically encoded biosensor probes can be used to make quantitative measurements of changes in the activity of cell signaling pathways. Then, we describe how fluorescence lifetime imaging can be used for label-free measurements of the metabolic states of cells within the living animal.

Original languageEnglish (US)
Pages (from-to)95-104
Number of pages10
JournalMethods
Volume128
DOIs
StatePublished - Sep 1 2017

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Keywords

  • Biosensor probes
  • Fluorescence lifetime imaging microscopy (FLIM)
  • Intravital microscopy (IVM)
  • Intrinsic fluorescence
  • Ratiometric imaging

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)

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