Intravital microscopy of biosensor activities and intrinsic metabolic states

Seth Winfree, Takashi Hato, Richard Day

Research output: Contribution to journalArticle

3 Citations (Scopus)

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)
JournalMethods
DOIs
StateAccepted/In press - Jan 26 2017

Fingerprint

Biosensing Techniques
Biosensors
Imaging techniques
Animals
Optical Imaging
Tissue
Cell signaling
Calibration
Labels
Fluorescence
Scattering
Intravital Microscopy

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)

Cite this

Intravital microscopy of biosensor activities and intrinsic metabolic states. / Winfree, Seth; Hato, Takashi; Day, Richard.

In: Methods, 26.01.2017.

Research output: Contribution to journalArticle

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