Widefield FLIM imaging with an ultra-sensitive single Photon Counting Camera

Session

Poster Session

Authors

André Weber (2), Yury Prokazov (4), Stefan Richter (2), Ezgi Altun (3), Rodrigo Herrera-Molina (1, 5), Torsten Stöter (2), Werner Zuschratter (2)

Affiliations

1. CIBQA - Universidad Bernardo O‘Higgins
2. Leibniz Institute for Neurobiology
3. Otto von Guericke University
4. Photonscore GmbH
5. GWU - The George Washington University

Keywords

Fluorescence Lifetime Imaging Microscopy (FLIM)

Single Photon Counting camera

Metabolic Imaging

Labe free Imaging

Multimodal Imaging

Abstract text

Current fluorescence microscopy often requires relatively high doses of light, which can harm living specimens due to photon damage. In this paper, we introduce an alternative approach using a wide-field imaging technique with a highly sensitive detector. Our system utilizes a position-sensitive detector with multichannel plates for electron amplification (LINCam, Photonscore GmbH), and operates based on single-photon counting. With new photocathodes boasting high quantum efficiency and exceptional signal-to-noise ratios, our system allows imaging under very low light conditions (< 30 mW/cm²), producing diffraction-limited FLIM images by capturing individual light quanta's position and time information in a continuous data stream. This method offers a positional accuracy akin to a 1000 × 1000 pixel camera with a time accuracy of 40 ps, facilitating long observation times for sensitive samples. Its applications span calcium imaging, single molecule detection, FLIM-FRET, and quantum optics. Additionally, it enables label-free imaging of metabolic processes by detecting autofluorescence from metabolites like NADH and FAD, promising new avenues for studying cellular and tissue vitality under pharmacological stress and distinguishing between healthy and diseased tissue in medical diagnostics. Moreover, the system can simultaneously gather multimodal data from multiple detectors and can be expanded with a spectrometer into a hyperspectral time-resolved Raman FLIM imaging system.

References

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