Light-field tomographic fluorescence lifetime imaging microscopy

Yayao Ma; Luzhe Huang; Chandani Sen; Samuel Burri; Claudio Bruschini; Xilin Yang; Robert B Cameron; Gregory A Fishbein; Brigitte N Gomperts; Aydogan Ozcan; Edoardo Charbon; Liang Gao

doi:10.21203/rs.3.rs-2883279/v1

Light-field tomographic fluorescence lifetime imaging microscopy

Res Sq [Preprint]. 2023 May 10:rs.3.rs-2883279. doi: 10.21203/rs.3.rs-2883279/v1.

Authors

Yayao Ma¹, Luzhe Huang^{1

2

3}, Chandani Sen⁴, Samuel Burri⁵, Claudio Bruschini⁵, Xilin Yang^{1

2

3}, Robert B Cameron⁶, Gregory A Fishbein⁷, Brigitte N Gomperts⁴, Aydogan Ozcan^{1

2

3

8}, Edoardo Charbon⁵, Liang Gao^{1

3}

Affiliations

¹ Department of Bioengineering, University of California, Los Angeles, CA, USA.
² Electrical and Computer Engineering Department, University of California, Los Angeles, CA, USA.
³ California Nano Systems Institute, University of California, Los Angeles, CA, USA.
⁴ UCLA Children's Discovery and Innovation Institute, Mattel Children's Hospital UCLA, Department of Pediatrics, David Geffen School of Medicine, UCLA, Los Angeles, CA, USA.
⁵ Advanced Quantum Architecture Laboratory, Ecole Polytechnique Federale de Lausanne, Neuchatel, Switzerland.
⁶ Department of Thoracic Surgery, University of California, Los Angeles, CA, USA.
⁷ Department of Pathology and Laboratory Medicine, David Geffen School of Medicine at UCLA, Los Angeles, CA, USA.
⁸ David Geffen School of Medicine, University of California Los Angeles, Los Angeles, CA, USA.

Abstract

Fluorescence lifetime imaging microscopy (FLIM) is a powerful imaging technique that enables the visualization of biological samples at the molecular level by measuring the fluorescence decay rate of fluorescent probes. This provides critical information about molecular interactions, environmental changes, and localization within biological systems. However, creating high-resolution lifetime maps using conventional FLIM systems can be challenging, as it often requires extensive scanning that can significantly lengthen acquisition times. This issue is further compounded in three-dimensional (3D) imaging because it demands additional scanning along the depth axis. To tackle this challenge, we developed a novel computational imaging technique called light field tomographic FLIM (LIFT-FLIM). Our approach allows for the acquisition of volumetric fluorescence lifetime images in a highly data-efficient manner, significantly reducing the number of scanning steps required compared to conventional point-scanning or line-scanning FLIM imagers. Moreover, LIFT-FLIM enables the measurement of high-dimensional data using low-dimensional detectors, which are typically low-cost and feature a higher temporal bandwidth. We demonstrated LIFT-FLIM using a linear single-photon avalanche diode array on various biological systems, showcasing unparalleled single-photon detection sensitivity. Additionally, we expanded the functionality of our method to spectral FLIM and demonstrated its application in high-content multiplexed imaging of lung organoids. LIFT-FLIM has the potential to open up new avenues in both basic and translational biomedical research.

Publication types

Preprint