Forward-collected simultaneous fluorescence lifetime imaging and coherent anti-Stokes Raman scattering microscopy

J Biomed Opt. 2011 Feb;16(2):021103. doi: 10.1117/1.3490641.

Abstract

We demonstrate the simultaneous collection and separation of femtosecond-laser-based forward-collected coherent anti-Stokes Raman scattering (F-CARS) and two-photon-excitation-induced fluorescence lifetime images (FLIM) using time-correlated single photon counting (TCSPC). We achieve this in a nondescanned geometry using a single multimode fiber without significant loss of light, field of view, and most importantly, TCSPC timing fidelity. In addition to showing the ability to separate CARS images from FLIM images using time gating, we also demonstrate composite multimodal epicollected FLIM imaging with fiber-collected F-CARS imaging in live cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Equipment Design
  • Equipment Failure Analysis
  • Image Enhancement / instrumentation*
  • Microscopy, Fluorescence / instrumentation*
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Spectrum Analysis, Raman / instrumentation*
  • Tomography, Optical Coherence / instrumentation*