High contrast imaging of reversibly switchable fluorescent proteins via temporally unmixed Multispectral Optoacoustic Tomography (tuMSOT)

TL;DR

This paper introduces a method using temporally unmixed multispectral optoacoustic tomography (tuMSOT) to enhance contrast and discriminate between similar fluorescent proteins in blood-rich environments, improving multiplexed molecular imaging.

Contribution

The study demonstrates the use of photocontrol and temporal unmixing in MSOT to distinguish similar RSFPs, advancing high-contrast, multiplexed optoacoustic imaging techniques.

Findings

Enhanced contrast-to-noise ratios for RSFPs in blood-rich environments.

Successful discrimination of similar RSFP variants using tuMSOT.

Improved multiplexing capabilities in molecular optoacoustic imaging.

Abstract

Photocontrol of reversibly switchable fluorescent proteins (RSFPs) was used to program optoacoustic signal time courses that were temporally unmixed to increase the proteins contrast-to-noise-ratios (CNRs) in optoacoustic imaging. In this way, two variants of the RSFP Dronpa with very similar optoacoustic spectra could be readily discriminated in the presence of highly absorbing blood. Addition of temporal unmixing to multispectral optoacoustic tomography (tuMSOT) in conjunction with synthetic or genetically controlled photochromic contrast agents and customized photoswitching schedules can increase the performance of multiplexed and high contrast molecular optoacoustic imaging.