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Wide-Field Mid-Infrared Hyperspectral Imaging by Snapshot Phase Contrast Measurement of Optothermal Excitation
[Image: see text] Vibrational microscopy methods based on Raman scattering or infrared absorption provide a label-free approach for chemical-contrast imaging, but employ point-by-point scanning and impose a compromise between the imaging speed and field-of-view (FOV). Optothermal microscopy has been...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical Society
2021
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8613735/ https://www.ncbi.nlm.nih.gov/pubmed/34766751 http://dx.doi.org/10.1021/acs.analchem.1c02805 |
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author | Yuan, Tao Pleitez, Miguel A. Gasparin, Francesca Ntziachristos, Vasilis |
author_facet | Yuan, Tao Pleitez, Miguel A. Gasparin, Francesca Ntziachristos, Vasilis |
author_sort | Yuan, Tao |
collection | PubMed |
description | [Image: see text] Vibrational microscopy methods based on Raman scattering or infrared absorption provide a label-free approach for chemical-contrast imaging, but employ point-by-point scanning and impose a compromise between the imaging speed and field-of-view (FOV). Optothermal microscopy has been proposed as a promising imaging modality to avoid this compromise, although at restrictively small FOVs capable of imaging only few cells. Here, we present wide-field optothermal mid-infrared microscopy (WOMiM) for wide-field chemical-contrast imaging based on snapshot pump–probe detection of optothermal signal, using a custom-made condenser-free phase contrast microscopy to capture the phase change of samples after mid-infrared irradiation. We achieved chemical contrast for FOVs up to 180 μm in diameter, yielding 10-fold larger imaging areas than the state-of-the-art, at imaging speeds of 1 ms/frame. The maximum possible imaging speed of WOMiM was determined by the relaxation time of optothermal heat, measured to be 32.8 μs in water, corresponding to a frame rate of ∼30 kHz. This proof-of-concept demonstrates that vibrational imaging can be achieved at an unprecedented imaging speed and large FOV with the potential to significantly facilitate label-free imaging of cellular dynamics. |
format | Online Article Text |
id | pubmed-8613735 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | American Chemical Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-86137352021-11-26 Wide-Field Mid-Infrared Hyperspectral Imaging by Snapshot Phase Contrast Measurement of Optothermal Excitation Yuan, Tao Pleitez, Miguel A. Gasparin, Francesca Ntziachristos, Vasilis Anal Chem [Image: see text] Vibrational microscopy methods based on Raman scattering or infrared absorption provide a label-free approach for chemical-contrast imaging, but employ point-by-point scanning and impose a compromise between the imaging speed and field-of-view (FOV). Optothermal microscopy has been proposed as a promising imaging modality to avoid this compromise, although at restrictively small FOVs capable of imaging only few cells. Here, we present wide-field optothermal mid-infrared microscopy (WOMiM) for wide-field chemical-contrast imaging based on snapshot pump–probe detection of optothermal signal, using a custom-made condenser-free phase contrast microscopy to capture the phase change of samples after mid-infrared irradiation. We achieved chemical contrast for FOVs up to 180 μm in diameter, yielding 10-fold larger imaging areas than the state-of-the-art, at imaging speeds of 1 ms/frame. The maximum possible imaging speed of WOMiM was determined by the relaxation time of optothermal heat, measured to be 32.8 μs in water, corresponding to a frame rate of ∼30 kHz. This proof-of-concept demonstrates that vibrational imaging can be achieved at an unprecedented imaging speed and large FOV with the potential to significantly facilitate label-free imaging of cellular dynamics. American Chemical Society 2021-11-12 2021-11-23 /pmc/articles/PMC8613735/ /pubmed/34766751 http://dx.doi.org/10.1021/acs.analchem.1c02805 Text en © 2021 The Authors. Published by American Chemical Society https://creativecommons.org/licenses/by-nc-nd/4.0/Permits non-commercial access and re-use, provided that author attribution and integrity are maintained; but does not permit creation of adaptations or other derivative works (https://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Yuan, Tao Pleitez, Miguel A. Gasparin, Francesca Ntziachristos, Vasilis Wide-Field Mid-Infrared Hyperspectral Imaging by Snapshot Phase Contrast Measurement of Optothermal Excitation |
title | Wide-Field Mid-Infrared
Hyperspectral Imaging by Snapshot
Phase Contrast Measurement of Optothermal Excitation |
title_full | Wide-Field Mid-Infrared
Hyperspectral Imaging by Snapshot
Phase Contrast Measurement of Optothermal Excitation |
title_fullStr | Wide-Field Mid-Infrared
Hyperspectral Imaging by Snapshot
Phase Contrast Measurement of Optothermal Excitation |
title_full_unstemmed | Wide-Field Mid-Infrared
Hyperspectral Imaging by Snapshot
Phase Contrast Measurement of Optothermal Excitation |
title_short | Wide-Field Mid-Infrared
Hyperspectral Imaging by Snapshot
Phase Contrast Measurement of Optothermal Excitation |
title_sort | wide-field mid-infrared
hyperspectral imaging by snapshot
phase contrast measurement of optothermal excitation |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8613735/ https://www.ncbi.nlm.nih.gov/pubmed/34766751 http://dx.doi.org/10.1021/acs.analchem.1c02805 |
work_keys_str_mv | AT yuantao widefieldmidinfraredhyperspectralimagingbysnapshotphasecontrastmeasurementofoptothermalexcitation AT pleitezmiguela widefieldmidinfraredhyperspectralimagingbysnapshotphasecontrastmeasurementofoptothermalexcitation AT gasparinfrancesca widefieldmidinfraredhyperspectralimagingbysnapshotphasecontrastmeasurementofoptothermalexcitation AT ntziachristosvasilis widefieldmidinfraredhyperspectralimagingbysnapshotphasecontrastmeasurementofoptothermalexcitation |