Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Yuan, Tao, Pleitez, Miguel A., Gasparin, Francesca, Ntziachristos, Vasilis
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2021
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
_version_ 1784603698345803776
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