Multiplexed Whole-animal Imaging with Reversibly Switchable Optoacoustic Proteins

Overview

Journal Sci Adv

Specialties Biology
Science

Date 2020 Jun 26

PMID 32582850

Citations 20

Authors

Kanuj Mishra

Mariia Stankevych

Juan Pablo Fuenzalida-Werner

Simon Grassmann

Vipul Gujrati

Yuanhui Huang

Uwe Klemm

Veit R Buchholz

Vasilis Ntziachristos

Andre C Stiel

Affiliations

Soon will be listed here.

Abstract

We introduce two photochromic proteins for cell-specific in vivo optoacoustic (OA) imaging with signal unmixing in the temporal domain. We show highly sensitive, multiplexed visualization of T lymphocytes, bacteria, and tumors in the mouse body and brain. We developed machine learning-based software for commercial imaging systems for temporal unmixed OA imaging, enabling its routine use in life sciences.

Citing Articles

Nanophotonic-enhanced photoacoustic imaging for brain tumor detection.

Rizwan A, Sridharan B, Park J, Kim D, Vial J, Kyhm K J Nanobiotechnology. 2025; 23(1):170.

PMID: 40045308 PMC: 11881315. DOI: 10.1186/s12951-025-03204-5.

Expanding the fluorescent toolkit: Blue fluorescent protein-expressing Plasmodium berghei for enhanced multiplex microscopy.

Atchou K, Caldelari R, Roques M, Schmuckli-Maurer J, Beyeler R, Heussler V PLoS One. 2025; 20(3):e0308055.

PMID: 40029851 PMC: 11875362. DOI: 10.1371/journal.pone.0308055.

Controlling the sound of light: photoswitching optoacoustic imaging.

Stiel A, Ntziachristos V Nat Methods. 2024; 21(11):1996-2007.

PMID: 39322752 DOI: 10.1038/s41592-024-02396-2.

Reversibly photoswitchable protein assemblies with collagen affinity for in vivo photoacoustic imaging of tumors.

Chen S, Li K, Chen X, Lei S, Lin J, Huang P Sci Adv. 2024; 10(35):eadn8274.

PMID: 39213344 PMC: 11364091. DOI: 10.1126/sciadv.adn8274.

Photoacoustic Tomography with Temporal Encoding Reconstruction (PATTERN) for cross-modal individual analysis of the whole brain.

Chen Y, Yang H, Luo Y, Niu Y, Yu M, Deng S Nat Commun. 2024; 15(1):4228.

PMID: 38762498 PMC: 11102525. DOI: 10.1038/s41467-024-48393-z.

References

Liu Y, Bhattarai P, Dai Z, Chen X . Photothermal therapy and photoacoustic imaging via nanotheranostics in fighting cancer. Chem Soc Rev. 2018; 48(7):2053-2108. PMC: 6437026. DOI: 10.1039/c8cs00618k. View

Mishra K, Fuenzalida-Werner J, Ntziachristos V, Stiel A . Photocontrollable Proteins for Optoacoustic Imaging. Anal Chem. 2019; 91(9):5470-5477. DOI: 10.1021/acs.analchem.9b01048. View

Li L, Shemetov A, Baloban M, Hu P, Zhu L, Shcherbakova D . Small near-infrared photochromic protein for photoacoustic multi-contrast imaging and detection of protein interactions in vivo. Nat Commun. 2018; 9(1):2734. PMC: 6048155. DOI: 10.1038/s41467-018-05231-3. View

Steinberg I, Huland D, Vermesh O, Frostig H, Tummers W, Gambhir S . Photoacoustic clinical imaging. Photoacoustics. 2019; 14:77-98. PMC: 6595011. DOI: 10.1016/j.pacs.2019.05.001. View

Chee R, Li Y, Zhang W, Campbell R, Zemp R . In vivo photoacoustic difference-spectra imaging of bacteria using photoswitchable chromoproteins. J Biomed Opt. 2018; 23(10):1-11. DOI: 10.1117/1.JBO.23.10.106006. View

Yang X, Kuk J, Moffat K . Crystal structure of Pseudomonas aeruginosa bacteriophytochrome: photoconversion and signal transduction. Proc Natl Acad Sci U S A. 2008; 105(38):14715-20. PMC: 2567202. DOI: 10.1073/pnas.0806718105. View

Bellini D, Papiz M . Structure of a bacteriophytochrome and light-stimulated protomer swapping with a gene repressor. Structure. 2012; 20(8):1436-46. DOI: 10.1016/j.str.2012.06.002. View

Yao J, Kaberniuk A, Li L, Shcherbakova D, Zhang R, Wang L . Multiscale photoacoustic tomography using reversibly switchable bacterial phytochrome as a near-infrared photochromic probe. Nat Methods. 2015; 13(1):67-73. PMC: 4697872. DOI: 10.1038/nmeth.3656. View

Stiel A, Dean-Ben X, Jiang Y, Ntziachristos V, Razansky D, Westmeyer G . High-contrast imaging of reversibly switchable fluorescent proteins via temporally unmixed multispectral optoacoustic tomography. Opt Lett. 2015; 40(3):367-70. DOI: 10.1364/OL.40.000367. View

10.

Wang L, Hu S . Photoacoustic tomography: in vivo imaging from organelles to organs. Science. 2012; 335(6075):1458-62. PMC: 3322413. DOI: 10.1126/science.1216210. View

11.

Ntziachristos V, Razansky D . Molecular imaging by means of multispectral optoacoustic tomography (MSOT). Chem Rev. 2010; 110(5):2783-94. DOI: 10.1021/cr9002566. View

12.

Yao J, Wang L, Yang J, Maslov K, Wong T, Li L . High-speed label-free functional photoacoustic microscopy of mouse brain in action. Nat Methods. 2015; 12(5):407-10. PMC: 4428901. DOI: 10.1038/nmeth.3336. View

13.

Gujrati V, Mishra A, Ntziachristos V . Molecular imaging probes for multi-spectral optoacoustic tomography. Chem Commun (Camb). 2017; 53(34):4653-4672. DOI: 10.1039/c6cc09421j. View

14.

Knieling F, Neufert C, Hartmann A, Claussen J, Urich A, Egger C . Multispectral Optoacoustic Tomography for Assessment of Crohn's Disease Activity. N Engl J Med. 2017; 376(13):1292-1294. DOI: 10.1056/NEJMc1612455. View

15.

Reber J, Willershauser M, Karlas A, Paul-Yuan K, Diot G, Franz D . Non-invasive Measurement of Brown Fat Metabolism Based on Optoacoustic Imaging of Hemoglobin Gradients. Cell Metab. 2018; 27(3):689-701.e4. DOI: 10.1016/j.cmet.2018.02.002. View

16.

Wagner J, Zhang J, von Stetten D, Gunther M, Murgida D, Mroginski M . Mutational analysis of Deinococcus radiodurans bacteriophytochrome reveals key amino acids necessary for the photochromicity and proton exchange cycle of phytochromes. J Biol Chem. 2008; 283(18):12212-26. PMC: 2431007. DOI: 10.1074/jbc.M709355200. View

17.

Wang L, Yao J . A practical guide to photoacoustic tomography in the life sciences. Nat Methods. 2016; 13(8):627-38. PMC: 4980387. DOI: 10.1038/nmeth.3925. View

18.

Vetschera P, Mishra K, Fuenzalida-Werner J, Chmyrov A, Ntziachristos V, Stiel A . Characterization of Reversibly Switchable Fluorescent Proteins in Optoacoustic Imaging. Anal Chem. 2018; 90(17):10527-10535. DOI: 10.1021/acs.analchem.8b02599. View

19.

Brunker J, Yao J, Laufer J, Bohndiek S . Photoacoustic imaging using genetically encoded reporters: a review. J Biomed Opt. 2017; 22(7). DOI: 10.1117/1.JBO.22.7.070901. View

20.

Hai P, Imai T, Xu S, Zhang R, Aft R, Zou J . High-throughput, label-free, single-cell photoacoustic microscopy of intratumoral metabolic heterogeneity. Nat Biomed Eng. 2019; 3(5):381-391. PMC: 6544054. DOI: 10.1038/s41551-019-0376-5. View