Time-resolved multifocal multiphoton microscope for high speed FRET imaging in vivo

Opt Lett. 2014 Oct 15;39(20):6013-6. doi: 10.1364/OL.39.006013.

Abstract

Imaging the spatiotemporal interaction of proteins in vivo is essential to understanding the complexities of biological systems. The highest accuracy monitoring of protein-protein interactions is achieved using Förster resonance energy transfer (FRET) measured by fluorescence lifetime imaging, with measurements taking minutes to acquire a single frame, limiting their use in dynamic live cell systems. We present a diffraction limited, massively parallel, time-resolved multifocal multiphoton microscope capable of producing fluorescence lifetime images with 55 ps time-resolution, giving improvements in acquisition speed of a factor of 64. We present demonstrations with FRET imaging in a model cell system and demonstrate in vivo FLIM using a GTPase biosensor in the zebrafish embryo.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Fluorescence Resonance Energy Transfer*
  • MCF-7 Cells
  • Microscopy, Fluorescence, Multiphoton / methods*
  • Time Factors
  • Zebrafish