Multimodal hyperspectral optical microscopy
Abstract
We describe a unique and convenient approach to multimodal hyperspectral optical microscopy, herein achieved by coupling a portable and transferable hyperspectral imager to various optical microscopes. The experimental and data analysis schemes involved in recording spectrally and spatially resolved fluorescence, dark field, and optical absorption micrographs are illustrated through prototypical measurements targeting selected model systems. Namely, hyperspectral fluorescence micrographs of isolated fluorescent beads are employed to ensure spectral calibration of our detector and to gauge the attainable spatial resolution of our measurements; the recorded images are diffraction-limited. Moreover, spatially over-sampled absorption spectroscopy of a single lipid (18:1 Liss Rhod PE) layer reveals that optical densities on the order of 10-3 may be resolved by spatially averaging the recorded optical signatures. We also briefly illustrate two applications of our setup in the general areas of plasmonics and cell biology. Most notably, we deploy hyperspectral optical absorption microscopy to identify and image algal pigments within a single live Tisochrysis lutea cell. Overall, this work paves the way for multimodal multidimensional spectral imaging measurements spanning the realms of several scientific disciples.
- Authors:
-
[1];
- Pacific Northwest National Lab. (PNNL), Richland, WA (United States). Environmental Molecular Sciences Lab. (EMSL)
- Pacific Northwest National Lab. (PNNL), Richland, WA (United States). Physical Sciences Div.
- Surface Optics Corp., San Diego, CA (United States)
- Publication Date:
- Research Org.:
- Pacific Northwest National Lab. (PNNL), Richland, WA (United States). Environmental Molecular Sciences Lab. (EMSL)
- Sponsoring Org.:
- USDOE Office of Science (SC), Biological and Environmental Research (BER); USDOE Office of Science (SC), Basic Energy Sciences (BES)
- OSTI Identifier:
- 1411908
- Alternate Identifier(s):
- OSTI ID: 1495725
- Report Number(s):
- PNNL-SA-120193
Journal ID: ISSN 0301-0104; 49643; KC0301050; KP1606000; KP1606000; TRN: US1800284
- Grant/Contract Number:
- AC05-76RL01830; AC05-76RL1830
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Chemical Physics
- Additional Journal Information:
- Journal Volume: 498-499; Journal ID: ISSN 0301-0104
- Publisher:
- Elsevier
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; 47 OTHER INSTRUMENTATION; Environmental Molecular Sciences Laboratory
Citation Formats
Novikova, Irina V., Smallwood, Chuck R., Gong, Yu, Hu, Dehong, Hendricks, Leif, Evans, James E., Bhattarai, Ashish, Hess, Wayne P., and El-Khoury, Patrick Z. Multimodal hyperspectral optical microscopy. United States: N. p., 2017.
Web. doi:10.1016/j.chemphys.2017.08.011.
Novikova, Irina V., Smallwood, Chuck R., Gong, Yu, Hu, Dehong, Hendricks, Leif, Evans, James E., Bhattarai, Ashish, Hess, Wayne P., & El-Khoury, Patrick Z. Multimodal hyperspectral optical microscopy. United States. https://doi.org/10.1016/j.chemphys.2017.08.011
Novikova, Irina V., Smallwood, Chuck R., Gong, Yu, Hu, Dehong, Hendricks, Leif, Evans, James E., Bhattarai, Ashish, Hess, Wayne P., and El-Khoury, Patrick Z. Sat .
"Multimodal hyperspectral optical microscopy". United States. https://doi.org/10.1016/j.chemphys.2017.08.011. https://www.osti.gov/servlets/purl/1411908.
@article{osti_1411908,
title = {Multimodal hyperspectral optical microscopy},
author = {Novikova, Irina V. and Smallwood, Chuck R. and Gong, Yu and Hu, Dehong and Hendricks, Leif and Evans, James E. and Bhattarai, Ashish and Hess, Wayne P. and El-Khoury, Patrick Z.},
abstractNote = {We describe a unique and convenient approach to multimodal hyperspectral optical microscopy, herein achieved by coupling a portable and transferable hyperspectral imager to various optical microscopes. The experimental and data analysis schemes involved in recording spectrally and spatially resolved fluorescence, dark field, and optical absorption micrographs are illustrated through prototypical measurements targeting selected model systems. Namely, hyperspectral fluorescence micrographs of isolated fluorescent beads are employed to ensure spectral calibration of our detector and to gauge the attainable spatial resolution of our measurements; the recorded images are diffraction-limited. Moreover, spatially over-sampled absorption spectroscopy of a single lipid (18:1 Liss Rhod PE) layer reveals that optical densities on the order of 10-3 may be resolved by spatially averaging the recorded optical signatures. We also briefly illustrate two applications of our setup in the general areas of plasmonics and cell biology. Most notably, we deploy hyperspectral optical absorption microscopy to identify and image algal pigments within a single live Tisochrysis lutea cell. Overall, this work paves the way for multimodal multidimensional spectral imaging measurements spanning the realms of several scientific disciples.},
doi = {10.1016/j.chemphys.2017.08.011},
journal = {Chemical Physics},
number = ,
volume = 498-499,
place = {United States},
year = {Sat Sep 02 00:00:00 EDT 2017},
month = {Sat Sep 02 00:00:00 EDT 2017}
}
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Works referenced in this record:
Multimodal hyperspectral optical microscopy
journal, November 2017
- Novikova, Irina V.; Smallwood, Chuck R.; Gong, Yu
- Chemical Physics, Vol. 498-499
Broadband single-molecule excitation spectroscopy
journal, January 2016
- Piatkowski, Lukasz; Gellings, Esther; van Hulst, Niek F.
- Nature Communications, Vol. 7, Issue 1