Correlative imaging to resolve molecular structures in individual cells: Substrate validation study for super-resolution infrared microspectroscopy

Light microscopy has been a favorite tool of biological studies for almost a century, recently producing detailed images with exquisite molecular specificity achieving spatial resolution at nanoscale. However, light microscopy is insufficient to provide chemical information as a standalone technique...

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Bibliographic Details
Published in:	Nanomedicine Vol. 43; p. 102563
Main Authors:	Paulus, Agnes, Yogarasa, Sahana, Kansiz, Mustafa, Martinsson, Isak, Gouras, Gunnar K., Deierborg, Tomas, Engdahl, Anders, Borondics, Ferenc, Klementieva, Oxana
Format:	Journal Article
Language:	English
Published:	United States Elsevier Inc 01-07-2022
Subjects:	Atom and Molecular Physics and Optics Atom- och molekylfysik och optik Brain Fysik Glass coverslips Microspectroscopy Natural Sciences Naturvetenskap Neuron Optical photothermal infrared Physical Sciences Single-cell analysis Brain O-PTIR Single-cell analysis AFM IR μIR hAβ(1–42) CTB Optical photothermal infrared Glass coverslips Neuron FTIR Microspectroscopy SEM
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Summary:	Light microscopy has been a favorite tool of biological studies for almost a century, recently producing detailed images with exquisite molecular specificity achieving spatial resolution at nanoscale. However, light microscopy is insufficient to provide chemical information as a standalone technique. An increasing amount of evidence demonstrates that optical photothermal infrared microspectroscopy (O-PTIR) is a valuable imaging tool that can extract chemical information to locate molecular structures at submicron resolution. To further investigate the applicability of sub-micron infrared microspectroscopy for biomedical applications, we analyzed the contribution of substrate chemistry to the infrared spectra acquired from individual neurons grown on various imaging substrates. To provide an example of correlative immunofluorescence/O-PTIR imaging, we used immunofluorescence to locate specific organelles for O-PTIR measurement, thus capturing molecular structures at the sub-cellular level directly in cells, which is not possible using traditional infrared microspectroscopy or immunofluorescence microscopy alone. Light microscopy has been a favorite tool of biological studies, but it is insufficient as a standalone technique to provide chemical information alongside high-resolution images. Here we analyzed various imaging substrates, addressing substrate chemistry contribution to the spectra acquired by optical photothermal infrared microspectroscopy (O-PTIR). O-PTIR is an innovative imaging tool that can extract chemical information and generate hyperspectral images that can be used to locate lipids, proteins, DNA, cell membranes, and other biomolecules at submicron resolution. We demonstrate that O-PTIR and immunofluorescence microscopy can be used to examine the same individual cells, providing correlated information at a subcellular level [Display omitted]
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	1549-9634 1549-9642 1549-9642
DOI:	10.1016/j.nano.2022.102563