Light-field tomographic fluorescence lifetime imaging microscopy

Area of Science:

• Biophotonics and Imaging Science
• Molecular and Cellular Biology
• Computational Imaging

Background:

• Fluorescence lifetime imaging microscopy (FLIM) visualizes biological samples at the molecular level by measuring fluorescence decay rates.
• Conventional FLIM faces challenges with long acquisition times and extensive scanning, especially for 3D imaging.
• High-resolution lifetime maps are crucial for understanding molecular interactions and cellular environments.

Purpose of the Study:

• To develop a computational imaging technique to overcome the limitations of conventional FLIM systems.
• To enable faster, more data-efficient acquisition of volumetric fluorescence lifetime images.
• To reduce the scanning requirements for high-resolution 3D FLIM.

Main Methods:

• Introduced light-field tomographic FLIM (LIFT-FLIM), a novel computational imaging approach.
• Utilized low-dimensional detectors with high temporal bandwidth for efficient data acquisition.
• Demonstrated LIFT-FLIM using a linear single-photon avalanche diode array.

Main Results:

• LIFT-FLIM significantly reduces scanning steps compared to traditional FLIM methods.
• Achieved highly data-efficient acquisition of volumetric fluorescence lifetime images.
• Showcased unparalleled single-photon detection sensitivity and expanded functionality to spectral FLIM.
• Applied LIFT-FLIM to high-content multiplexed imaging of lung organoids.

Conclusions:

• LIFT-FLIM offers a powerful solution for rapid, high-resolution 3D fluorescence lifetime imaging.
• The technique enhances molecular-level visualization in biological systems.
• LIFT-FLIM holds significant potential for advancing both basic and translational biomedical research.