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Fluorescently Guided Optical Photothermal Infrared Microspectroscopy for Protein-Specific Bioimaging at Subcellular Level

Prater, Craig ; Bai, Yeran ; Konings, Sabine C LU orcid ; Martinsson, Isak LU ; Swaminathan, Vinay S LU ; Nordenfelt, Pontus LU orcid ; Gouras, Gunnar LU orcid ; Borondics, Ferenc and Klementieva, Oxana LU orcid (2023) In Journal of Medicinal Chemistry 66(4). p.2542-2549
Abstract

Infrared spectroscopic imaging is widely used for the visualization of biomolecule structures, and techniques such as optical photothermal infrared (OPTIR) microspectroscopy can achieve <500 nm spatial resolution. However, these approaches lack specificity for particular cell types and cell components and thus cannot be used as a stand-alone technique to assess their properties. Here, we have developed a novel tool, fluorescently guided optical photothermal infrared microspectroscopy, that simultaneously exploits epifluorescence imaging and OPTIR to perform fluorescently guided IR spectroscopic analysis. This novel approach exceeds the diffraction limit of infrared microscopy and allows structural analysis of specific proteins... (More)

Infrared spectroscopic imaging is widely used for the visualization of biomolecule structures, and techniques such as optical photothermal infrared (OPTIR) microspectroscopy can achieve <500 nm spatial resolution. However, these approaches lack specificity for particular cell types and cell components and thus cannot be used as a stand-alone technique to assess their properties. Here, we have developed a novel tool, fluorescently guided optical photothermal infrared microspectroscopy, that simultaneously exploits epifluorescence imaging and OPTIR to perform fluorescently guided IR spectroscopic analysis. This novel approach exceeds the diffraction limit of infrared microscopy and allows structural analysis of specific proteins directly in tissue and single cells. Experiments described herein used epifluorescence to rapidly locate amyloid proteins in tissues or neuronal cultures, thus guiding OPTIR measurements to assess amyloid structures at the subcellular level. We believe that this new approach will be a valuable addition to infrared spectroscopy providing cellular specificity of measurements in complex systems for studies of structurally altered protein aggregates.

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author
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organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Medicinal Chemistry
volume
66
issue
4
pages
2542 - 2549
publisher
The American Chemical Society (ACS)
external identifiers
  • pmid:36599042
  • scopus:85146027211
ISSN
1520-4804
DOI
10.1021/acs.jmedchem.2c01359
language
English
LU publication?
yes
id
7766bb51-580b-4f30-bdff-98bac5315a5a
date added to LUP
2023-02-03 04:41:09
date last changed
2024-12-14 11:57:05
@article{7766bb51-580b-4f30-bdff-98bac5315a5a,
  abstract     = {{<p>Infrared spectroscopic imaging is widely used for the visualization of biomolecule structures, and techniques such as optical photothermal infrared (OPTIR) microspectroscopy can achieve &lt;500 nm spatial resolution. However, these approaches lack specificity for particular cell types and cell components and thus cannot be used as a stand-alone technique to assess their properties. Here, we have developed a novel tool, fluorescently guided optical photothermal infrared microspectroscopy, that simultaneously exploits epifluorescence imaging and OPTIR to perform fluorescently guided IR spectroscopic analysis. This novel approach exceeds the diffraction limit of infrared microscopy and allows structural analysis of specific proteins directly in tissue and single cells. Experiments described herein used epifluorescence to rapidly locate amyloid proteins in tissues or neuronal cultures, thus guiding OPTIR measurements to assess amyloid structures at the subcellular level. We believe that this new approach will be a valuable addition to infrared spectroscopy providing cellular specificity of measurements in complex systems for studies of structurally altered protein aggregates.</p>}},
  author       = {{Prater, Craig and Bai, Yeran and Konings, Sabine C and Martinsson, Isak and Swaminathan, Vinay S and Nordenfelt, Pontus and Gouras, Gunnar and Borondics, Ferenc and Klementieva, Oxana}},
  issn         = {{1520-4804}},
  language     = {{eng}},
  month        = {{01}},
  number       = {{4}},
  pages        = {{2542--2549}},
  publisher    = {{The American Chemical Society (ACS)}},
  series       = {{Journal of Medicinal Chemistry}},
  title        = {{Fluorescently Guided Optical Photothermal Infrared Microspectroscopy for Protein-Specific Bioimaging at Subcellular Level}},
  url          = {{http://dx.doi.org/10.1021/acs.jmedchem.2c01359}},
  doi          = {{10.1021/acs.jmedchem.2c01359}},
  volume       = {{66}},
  year         = {{2023}},
}