Deep-tissue SWIR imaging using rationally designed small red-shifted near-infrared fluorescent protein

Olena S Oliinyk; Chenshuo Ma; Sergei Pletnev; Mikhail Baloban; Carlos Taboada; Huaxin Sheng; Junjie Yao; Vladislav V Verkhusha

doi:10.1038/s41592-022-01683-0

Deep-tissue SWIR imaging using rationally designed small red-shifted near-infrared fluorescent protein

Nat Methods. 2023 Jan;20(1):70-74. doi: 10.1038/s41592-022-01683-0. Epub 2022 Dec 1.

Authors

Olena S Oliinyk¹, Chenshuo Ma², Sergei Pletnev³, Mikhail Baloban⁴, Carlos Taboada², Huaxin Sheng⁵, Junjie Yao⁶, Vladislav V Verkhusha^{7

8}

Affiliations

¹ Medicum, Faculty of Medicine, University of Helsinki, Helsinki, Finland.
² Department of Biomedical Engineering, School of Engineering, Duke University, Durham, NC, USA.
³ Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.
⁴ Department of Genetics and Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine, Bronx, NY, USA.
⁵ Department of Anesthesiology, School of Medicine, Duke University, Durham, NC, USA.
⁶ Department of Biomedical Engineering, School of Engineering, Duke University, Durham, NC, USA. junjie.yao@duke.edu.
⁷ Medicum, Faculty of Medicine, University of Helsinki, Helsinki, Finland. vladislav.verkhusha@einsteinmed.edu.
⁸ Department of Genetics and Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine, Bronx, NY, USA. vladislav.verkhusha@einsteinmed.edu.

Abstract

Applying rational design, we developed 17 kDa cyanobacteriochrome-based near-infrared (NIR-I) fluorescent protein, miRFP718nano. miRFP718nano efficiently binds endogenous biliverdin chromophore and brightly fluoresces in mammalian cells and tissues. miRFP718nano has maximal emission at 718 nm and an emission tail in the short-wave infrared (SWIR) region, allowing deep-penetrating off-peak fluorescence imaging in vivo. The miRFP718nano structure reveals the molecular basis of its red shift. We demonstrate superiority of miRFP718nano-enabled SWIR imaging over NIR-I imaging of microbes in the mouse digestive tract, mammalian cells injected into the mouse mammary gland and NF-kB activity in a mouse model of liver inflammation.

Publication types

Research Support, N.I.H., Intramural
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Fluorescent Dyes* / chemistry
Mammals
Mice
Optical Imaging*

Substances

Fluorescent Dyes

Abstract

Publication types

MeSH terms

Substances

Grants and funding