Label-Free High-Resolution Photothermal Optical Infrared Spectroscopy for Spatiotemporal Chemical Analysis in Fresh, Hydrated Living Tissues and Embryos

Nika Gvazava, Sabine C. Konings, Efrain Cepeda-Prado, Valeriia Skoryk, Chimezie H. Umeano, Jiao Dong, Iran A. N. Silva, Daniella Rylander Ottosson, Nicholas D. Leigh, Darcy Elizabeth Wagner, Oxana Klementieva

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

Label-free chemical imaging of living and functioning systems is the holy grail of biochemical research. However, existing techniques often require extensive sample preparation to remove interfering molecules such as water, rendering many molecular imaging techniques unsuitable for in situ structural studies. Here, we examined freshly extracted tissue biopsies and living small vertebrates at submicrometer resolution using optical photothermal infrared (O-PTIR) microspectroscopy and demonstrated the following major advances: (1) O-PTIR can be used for submicrometer structural analysis of unprocessed, fully hydrated tissue biopsies extracted from diverse organs, including living brain and lung tissues. (2) O-PTIR imaging can be performed on living organisms, such as salamander embryos, without compromising their further development. (3) Using O-PTIR, we tracked the structural changes of amyloids in functioning brain tissues over time, observing the appearance of newly formed amyloids for the first time. (4) Amyloid structures appeared altered following standard fixation and dehydration procedures. Thus, we demonstrate that O-PTIR enables time-resolved submicrometer in situ investigation of chemical and structural changes in diverse biomolecules in their native conditions, representing a technological breakthrough for in situ molecular imaging of biological samples.
Original languageEnglish
JournalJournal of the American Chemical Society
DOIs
Publication statusPublished - 2023
Externally publishedYes

Funding

This research was funded by grants to O.K.: Swedish Research Council Starting Grant #2021–03149; Åke Wibergs Stiftelse, Grant #M21–0146; and Swedish Brain Foundation, Grant # FO2022–0329, NanoLund support. N.D.L. is supported by the Knut and Alice Wallenberg Foundation and the Swedish Research Council, Grant #2020–01486. The Knut and Alice Wallenberg Foundation, the Medical Faculty at Lund University, and Region Skåne are acknowledged for financial support (D.E.W.). This project has received funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation program (grant agreement no. 805361) (D.E.W.). Lund University and the Strategic Research Area “Emerging Research Topics” are also acknowledged for support (D.E.W. and O.K.). The Swedish Foundation for Strategic Research (grant agreement no. UKR22–0081) is acknowledged for financial support (V.S.).

FundersFunder number
Medical Faculty at Lund University
Åke Wiberg Stiftelse21–0146
Horizon 2020 Framework Programme805361
European Research Council
Stiftelsen för Strategisk ForskningUKR22–0081
HjärnfondenFO2022–0329
Knut och Alice Wallenbergs Stiftelse
Vetenskapsrådet2020–01486

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