Hyperspectral fluorescence imaging using a high-speed silicon photomultiplier array

TL;DR

This paper introduces a high-speed, low-cost hyperspectral imaging system using silicon photomultiplier arrays, enabling rapid, multiplexed fluorescence imaging with spectral unmixing capabilities.

Contribution

It presents a novel hyperspectral detection system based on silicon photomultiplier arrays that achieves high-speed, high-throughput imaging with spectral unmixing in a conventional microscope.

Findings

Achieved 16 spectral channels at 50 MP/s (800 million spectra/sec)

Enabled multiplexing of multiple contrast agents over large areas

Detected subtle spectral shifts linked to molecular binding

Abstract

High-speed multiplex imaging of fluorescent probes is limited by a combination of spectral resolution, sensitivity, high cost and low light throughput of detectors, and filters. In this work, we present a hyperspectral detection system based on a silicon photomultiplier array that enables high-speed, high-light throughput hyperspectral imaging at low cost. We demonstrate 16 spectral channel imaging at 50 MP/s (800M spectra per second) with a conventional two photon microscope combined with a generalized spectral unmixing model that enables extraction of spectrally overlapping fluorophores. We show that the high spectral resolution combined with high throughput enables the multiplexing of multiple contrast agents over large areas and the detection of subtle spectral shifts associated with molecular binding. Silicon photomultiplier arrays may be a promising method to extend multiplex fluorescence imaging in a variety of scenarios.