PuSH - Publication Server of Helmholtz Zentrum München: Phase-shifting optothermal microscopy enables live-cell mid-infrared hyperspectral imaging of large cell populations at high confluency.

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Yuan, T. ; Riobo, L. ; Gasparin, F. ; Ntziachristos, V. ; Pleitez, M.A.

Phase-shifting optothermal microscopy enables live-cell mid-infrared hyperspectral imaging of large cell populations at high confluency.

Sci. Adv. 10:eadj7944 (2024)

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Rapid live-cell hyperspectral imaging at large fields of view (FOVs) and high cell confluency remains challenging for conventional vibrational spectroscopy-based microscopy technologies. At the same time, imaging at high cell confluency and large FOVs is important for proper cell function and statistical significance of measurements, respectively. Here, we introduce phase-shifting mid-infrared optothermal microscopy (PSOM), which interprets molecular-vibrational information as the optical path difference induced by mid-infrared absorption and can take snapshot vibrational images over broad excitation areas at high live-cell confluency. By means of phase-shifting, PSOM suppresses noise to a quarter of current optothermal microscopy modalities to allow capturing live-cell vibrational images at FOVs up to 50 times larger than state of the art. PSOM also reduces illumination power flux density (PFD) down to four orders of magnitude lower than other conventional vibrational microscopy methods, such as coherent anti-Stokes Raman scattering (CARS), thus considerably decreasing the risk of cell photodamage.

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Publication type Article: Journal article

Document type Scientific Article

Corresponding Author

Keywords Stimulated Raman-scattering; Band

ISSN (print) / ISBN 2375-2548

e-ISSN 2375-2548

Journal Science Advances

Quellenangaben Volume: 10, Issue: 8, Article Number: eadj7944

Publisher American Association for the Advancement of Science (AAAS)

Publishing Place Washington, DC [u.a.]

Non-patent literature Publications

Reviewing status Peer reviewed

Institute(s) Institute of Biological and Medical Imaging (IBMI)

Grants European Research Council (ERC) through the European Union's Horizon 2020 research and innovation programme
European Union
Helmholtz Munich Innovation and Translation Call (Opto-G)
Deutsche Forschungsgemeinschaft (DFG)