Characterization of Reversibly Switchable Fluorescent Proteins in Optoacoustic Imaging

Vetschera P, Mishra K, Fuenzalida-Werner JP, Chmyrov A, Ntziachristos V, Stiel AC (2018)

Publication Type: Journal article

Publication year: 2018

Journal

Analytical Chemistry American Chemical Society

Book Volume: 90

Pages Range: 10527-10535

Journal Issue: 17

DOI: 10.1021/acs.analchem.8b02599

Abstract

Reversibly switchable fluorescent proteins (rsFPs) have had a revolutionizing effect on life science imaging due to their contribution to sub-diffraction-resolution optical microscopy (nanoscopy). Initial studies showed that their use as labels could also be highly beneficial for emerging photo- or optoacoustic imaging. It could be shown that their use in optoacoustics (i) strongly improves the imaging contrast-to-noise ratio due to modulation and locked-in detection, (ii) facilitates fluence calibration, affording precise measurements of physiological parameters, and finally (iii) could boost spatial resolution following similar concepts as used for nanoscopy. However, rsFPs show different photophysical behavior in optoacoustics than in optical microscopy because optoacoustics requires pulsed illumination and depends on signal generation via nonradiative energy decay channels. This implies that rsFPs optimized for fluorescence imaging may not be ideal for optoacoustics. Here, we analyze the photophysical behavior of a broad range of rsFPs with optoacoustics and analyze how the experimental factors central to optoacoustic imaging influence the different types of rsFPs. Finally, we discuss how knowledge of the switching behavior can be exploited for various optoacoustic imaging approaches using sophisticated temporal unmixing schemes.

Involved external institutions

Helmholtz Zentrum München - Deutsches Forschungszentrum für Gesundheit und Umwelt (HMGU) / Helmholtz Munich DE Germany (DE)

How to cite

APA:

Vetschera, P., Mishra, K., Fuenzalida-Werner, J.P., Chmyrov, A., Ntziachristos, V., & Stiel, A.C. (2018). Characterization of Reversibly Switchable Fluorescent Proteins in Optoacoustic Imaging. Analytical Chemistry, 90(17), 10527-10535. https://doi.org/10.1021/acs.analchem.8b02599

MLA:

Vetschera, Paul, et al. "Characterization of Reversibly Switchable Fluorescent Proteins in Optoacoustic Imaging." Analytical Chemistry 90.17 (2018): 10527-10535.

BibTeX: Download