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Autofluorescence Imaging to Evaluate Cellular Metabolism
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Label-Free Metabolic Imaging In Vivo by Two-Photon Fluorescence Lifetime Endomicroscopy.

Wenxuan Liang1, Defu Chen2, Honghua Guan3

  • 1School of Biomedical Engineering, Suzhou Institute for Advanced Research, University of Science and Technology of China, Suzhou, Jiangsu 215000, China; School of Physical Sciences, University of Science and Technology of China, Hefei, Anhui 230026, China.

ACS Photonics
|December 27, 2024
PubMed
Summary

We developed a novel two-photon fluorescence lifetime imaging endomicroscope (2p-FLeM) for cellular metabolic profiling. This compact device reliably tracks NADH lifetime dynamics in vivo, enabling label-free functional histology for clinical applications.

Keywords:
NADH lifetime imagingfluorescence lifetime imaging microscopy (FLIM)label-free imagingmetabolic imagingtwo-photon endomicroscopy

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Area of Science:

  • Biomedical Optics
  • Microscopy
  • Cellular Metabolism

Background:

  • NADH fluorescence lifetime is a key biomarker for cellular metabolic status.
  • Existing NADH fluorescence lifetime imaging microscopy (FLIM) requires compact, flexible endomicroscopic systems for in vivo applications.

Purpose of the Study:

  • To develop a compact, high-resolution two-photon fluorescence lifetime imaging endomicroscope (2p-FLeM).
  • To demonstrate the system's capability for reliable in vivo metabolic profiling of living tissues.

Main Methods:

  • Development of a 2 mm diameter 2p-FLeM system with subcellular resolution.
  • Utilized safe excitation power (~30 mW) and moderate pixel dwelling time (≤10 μs).
  • Performed in vivo experiments on cultured cancer cells, mouse tumor models, and mouse kidneys.

Main Results:

  • The 2p-FLeM system reliably extracted NADH lifetime parameters from living tissues.
  • Successfully tracked NADH lifetime dynamics in cancer cells, tumors undergoing apoptosis, and kidneys during ischemia-reperfusion.
  • Achieved complementary structural and metabolic information without labels.

Conclusions:

  • The 2p-FLeM system offers a compact and efficient solution for in vivo NADH lifetime imaging.
  • Enables label-free functional histological imaging of internal organs.
  • Shows promise for clinical diagnosis and therapeutic monitoring.