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Ultrafast Widefield Mid-Infrared Photothermal Heterodyne Imaging
[Image: see text] Mid-infrared photothermal (MIP) microscopy is a valuable tool for sensitive and fast chemical imaging with high spatial resolution beyond the mid-infrared diffraction limit. The highest sensitivity is usually achieved with heterodyne MIP employing photodetector point-scans and lock...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical Society
2022
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9583073/ https://www.ncbi.nlm.nih.gov/pubmed/36197677 http://dx.doi.org/10.1021/acs.analchem.2c02548 |
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author | Paiva, Eduardo M. Schmidt, Florian M. |
author_facet | Paiva, Eduardo M. Schmidt, Florian M. |
author_sort | Paiva, Eduardo M. |
collection | PubMed |
description | [Image: see text] Mid-infrared photothermal (MIP) microscopy is a valuable tool for sensitive and fast chemical imaging with high spatial resolution beyond the mid-infrared diffraction limit. The highest sensitivity is usually achieved with heterodyne MIP employing photodetector point-scans and lock-in detection, while the fastest systems use camera-based widefield MIP with pulsed probe light. One challenge is to simultaneously achieve high sensitivity, spatial resolution, and speed in a large field of view. Here, we present widefield mid-infrared photothermal heterodyne (WIPH) imaging, where a digital frequency-domain lock-in (DFdLi) filter is used for simultaneous multiharmonic demodulation of MIP signals recorded by individual camera pixels at frame rates up to 200 kHz. The DFdLi filter enables the use of continuous-wave probe light, which, in turn, eliminates the need for synchronization schemes and allows measuring MIP decay curves. The WIPH approach is characterized by imaging potassium ferricyanide microparticles and applied to detect lipid droplets (alkyne-palmitic acid) in 3T3-L1 fibroblast cells, both in the cell-silent spectral region around 2100 cm(–1) using an external-cavity quantum cascade laser. The system achieved up to 4000 WIPH images per second at a signal-to-noise ratio of 5.52 and 1 μm spatial resolution in a 128 × 128 μm field of view. The technique opens up for real-time chemical imaging of fast processes in biology, medicine, and material science. |
format | Online Article Text |
id | pubmed-9583073 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | American Chemical Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-95830732022-10-21 Ultrafast Widefield Mid-Infrared Photothermal Heterodyne Imaging Paiva, Eduardo M. Schmidt, Florian M. Anal Chem [Image: see text] Mid-infrared photothermal (MIP) microscopy is a valuable tool for sensitive and fast chemical imaging with high spatial resolution beyond the mid-infrared diffraction limit. The highest sensitivity is usually achieved with heterodyne MIP employing photodetector point-scans and lock-in detection, while the fastest systems use camera-based widefield MIP with pulsed probe light. One challenge is to simultaneously achieve high sensitivity, spatial resolution, and speed in a large field of view. Here, we present widefield mid-infrared photothermal heterodyne (WIPH) imaging, where a digital frequency-domain lock-in (DFdLi) filter is used for simultaneous multiharmonic demodulation of MIP signals recorded by individual camera pixels at frame rates up to 200 kHz. The DFdLi filter enables the use of continuous-wave probe light, which, in turn, eliminates the need for synchronization schemes and allows measuring MIP decay curves. The WIPH approach is characterized by imaging potassium ferricyanide microparticles and applied to detect lipid droplets (alkyne-palmitic acid) in 3T3-L1 fibroblast cells, both in the cell-silent spectral region around 2100 cm(–1) using an external-cavity quantum cascade laser. The system achieved up to 4000 WIPH images per second at a signal-to-noise ratio of 5.52 and 1 μm spatial resolution in a 128 × 128 μm field of view. The technique opens up for real-time chemical imaging of fast processes in biology, medicine, and material science. American Chemical Society 2022-10-05 2022-10-18 /pmc/articles/PMC9583073/ /pubmed/36197677 http://dx.doi.org/10.1021/acs.analchem.2c02548 Text en © 2022 The Authors. Published by American Chemical Society https://creativecommons.org/licenses/by/4.0/Permits the broadest form of re-use including for commercial purposes, provided that author attribution and integrity are maintained (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Paiva, Eduardo M. Schmidt, Florian M. Ultrafast Widefield Mid-Infrared Photothermal Heterodyne Imaging |
title | Ultrafast Widefield
Mid-Infrared Photothermal Heterodyne
Imaging |
title_full | Ultrafast Widefield
Mid-Infrared Photothermal Heterodyne
Imaging |
title_fullStr | Ultrafast Widefield
Mid-Infrared Photothermal Heterodyne
Imaging |
title_full_unstemmed | Ultrafast Widefield
Mid-Infrared Photothermal Heterodyne
Imaging |
title_short | Ultrafast Widefield
Mid-Infrared Photothermal Heterodyne
Imaging |
title_sort | ultrafast widefield
mid-infrared photothermal heterodyne
imaging |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9583073/ https://www.ncbi.nlm.nih.gov/pubmed/36197677 http://dx.doi.org/10.1021/acs.analchem.2c02548 |
work_keys_str_mv | AT paivaeduardom ultrafastwidefieldmidinfraredphotothermalheterodyneimaging AT schmidtflorianm ultrafastwidefieldmidinfraredphotothermalheterodyneimaging |