High-throughput hyperspectral fluorescence imaging using a high-speed silicon photomultiplier array.

Area of Science:

• Biophotonics
• Microscopy
• Spectroscopy

Background:

• High-speed multiplex imaging of fluorescent probes faces limitations in spectral resolution, sensitivity, cost, and detector light throughput.
• Existing systems struggle to efficiently separate and detect spectrally overlapping fluorophores.

Purpose of the Study:

• To develop a cost-effective hyperspectral detection system for high-speed multiplex fluorescence imaging.
• To overcome the limitations of current systems by enhancing spectral resolution and light throughput.

Main Methods:

• Implementation of a hyperspectral detection system utilizing a silicon photomultiplier array.
• Integration with a conventional two-photon microscope.
• Application of a generalized spectral unmixing model for fluorophore separation.

Main Results:

• Demonstration of 16 spectral channel imaging at 50 MP/s (800M spectra/second).
• Successful extraction of spectrally overlapping fluorophores.
• Enabled multiplexing of multiple contrast agents and detection of subtle spectral shifts.

Conclusions:

• Silicon photomultiplier arrays offer a promising, low-cost solution for high-speed, high-throughput hyperspectral imaging.
• The developed system significantly advances multiplex fluorescence imaging capabilities.
• Potential for broad application in various biological and chemical sensing scenarios.