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Updated: May 15, 2025
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Author Spotlight: Standardizing Spheroid Formation Methods for Metabolic and Oxygenation Analysis Using Fluorescence Lifetime Imaging Microscopy
Published on: August 9, 2024
725
Advanced fluorescence lifetime-enhanced multiplexed nanoscopy of cells.
Samrat Basak1,2, Roman Tsukanov1
1Third Institute of Physics-Biophysics, Georg August University, 37077 Göttingen, Germany.
Methods in Microscopy
|April 7, 2025
Summary
This review highlights advancements in wide-field fluorescence lifetime imaging microscopy (FLIM) using the LINCam system. It enables single-molecule sensitivity for diverse bioimaging applications, including super-resolution microscopy.
Area of Science:
- Biophotonics and advanced microscopy techniques.
- Development of novel imaging instrumentation.
Background:
- Conventional intensity-based imaging lacks detailed molecular information.
- Fluorescence lifetime imaging microscopy (FLIM) provides richer data for biological studies.
- Wide-field FLIM offers high sensitivity and speed.
Purpose of the Study:
- To review significant advancements in wide-field FLIM.
- To showcase the capabilities of the LINCam system for single-molecule sensitivity.
- To detail applications in bioimaging and super-resolution microscopy.
Main Methods:
- Utilizing time-correlated single-photon counting (TCSPC) with a position-sensitive LINCam detector.
- Implementing wide-field FLIM for enhanced sensitivity and speed.
- Exploring applications in far-red single-molecule FLIM and super-resolution imaging.
Main Results:
- Demonstrated single-molecule sensitivity in wide-field FLIM.
- Enabled fast and multiplexed super-resolution imaging of cellular structures.
- Achieved high axial localization precision for 3D super-resolution imaging.
- Highlighted the potential of position-sensitive detectors for high-throughput imaging.
Conclusions:
- FLIM provides crucial information beyond intensity, expanding bioimaging possibilities.
- The LINCam system represents a significant step forward for sensitive, high-resolution microscopy.
- Future advancements in detector technology promise further breakthroughs in super-resolution imaging.
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