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

Paulus, Agnes; Yogarasa, Sahana; Kansiz, Mustafa; Martinsson, Isak; Gouras, Gunnar K.; Deierborg, Tomas; Engdahl, Anders; Borondics, Ferenc; Klementieva, Oxana

doi:10.1016/j.nano.2022.102563

Cited by 8 publications

(16 citation statements)

References 41 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In the third experiment (Figure ), we investigated the applicability of FL-OPTIR for the structural imaging of Aβ (1–42) in cultured primary neurons. Similar to tissue studies, the structural heterogeneity of amyloid proteins makes it challenging to understand the relationship between neurotoxicity and the structure of Aβ; therefore, to assess amyloid structures directly in neurons, OPTIR is the technique that can provide structural information at subcellular resolution. ,,, Figure shows that FL-OPTIR can be used to guide spectroscopic measurements based on fluorescent signals and unambiguously discriminate between Aβ and other proteins (Figures a and S7). Moreover, we show that FL-OPTIR can be used to assess Aβ structures associated with neurites (Figure b).…”

Section: Resultsmentioning

confidence: 99%

“…The spatial resolution of the OPTIR approach is also constant for all IR wavelengths, whereas traditional IR spectroscopy has a wavelength-dependent spatial resolution that worsens at long wavelengths/low wavenumbers. Although the elastic scattering is limited to the surface, the penetration depth of the OPTIR is defined by the reach of the IR beam (a few microns in the case of tissue). Note that the FL-OPTIR implementation developed herein is distinct from prior studies by other groups that are based on IR detection of thermosensitive fluorophores, IR and two-photon excited fluorescence, and OPTIR used for temperature-dependent changes in fluorescence quantum efficiency .…”

Section: Introductionmentioning

confidence: 99%

“…Specifically, infrared (IR) absorption induces a photothermal response consisting of thermal expansion and a change in the index of refraction of the sample, which leads to an intensity change of the probe light and allows the detection of the photothermal response. The wavelength of the used probe beam is 532 nm, which is smaller than the wavelength of absorbed IR light (3–20 μm), thus providing a 5–10 times improvement of the spatial resolution . Similar to conventional infrared spectroscopy, OPTIR provides information about the molecular bonds and, thus, can be used to characterize the chemical composition of the sample to study protein structures.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

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

Prater¹,

Bai

Konings

et al. 2023

J. Med. Chem.

Self Cite

View full text Add to dashboard Cite

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.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

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

Prater¹,

Bai

Konings

et al. 2023

J. Med. Chem.

Self Cite

View full text Add to dashboard Cite

show abstract

“…The fact that photothermal responses are measured with visible light wavelengths results in 5-10 times better resolution than traditional IR microspectroscopy techniques such as FTIR and QCL microscopies 8 . Importantly, when used in reflectance mode, it becomes a surface-scanning technique and therefore substrate chemistry (critical in transmission based IR methods) becomes negligible and perhaps more importantly, sample thickness becomes irrelevant 15 . While O-PTIR has been successfully used for the analysis of hydrated cells 18 and nematodes 8 , it is not yet known whether it can be applied for the analysis of living tissues and organisms without compromising their viability.…”

Section: Resultsmentioning

confidence: 99%

“…that is currently revolutionizing spatial analysis of materials [8][9][10][11][12][13][14][15][16][17] . O-PTIR uses two co-linear beams of light (IR and a visible probe beam) to measure photothermal responses of the sample by assessing intensity changes of the reflected (or transmitted) probe beam at 532 nm.…”

Section: Optical Photothermal Infrared Spectroscopy (O-ptir) Is An Em...mentioning

confidence: 99%

Label-free high-resolution infrared spectroscopy for spatiotemporal analysis of complex living systems

Gvazava

Konings

Cepeda-Prado

et al. 2023

Preprint

Self Cite

View full text Add to dashboard Cite

Label-free chemical and structural imaging of complex living tissue and biological systems is the holy grail of biomedical research and clinical diagnostics. The current analysis techniques are time-consuming and/or require extensive sample preparation, often due to the presence of interfering molecules such as water, making them unsuitable for the analysis of such systems. Here, we demonstrate a proof-of-principle study using label-free optical photothermal mid-infrared microspectroscopy (O-PTIR) for fast, direct spatiotemporal chemical analysis of complex living biological systems at submicron resolution. While other analytical methods can provide only static snapshots of molecular structures, our O-PTIR approach enables time-resolved and in situ investigation of chemical and structural changes of diverse biomolecules in their native conditions. This comprises a technological breakthrough in infrared spectroscopy to analyze biomolecules under native conditions over time: in fresh unprocessed biopsies, living brain tissue, and vertebrates without compromising their viability.One-Sentence SummaryProof-of-principle application of non-destructive O-PTIR for high-resolution spatiotemporal chemical and structural analysis of unprocessed biopsies, living brain tissue, and vertebrates.

show abstract

Bioactive Suture with Added Innate Defense Functionality for the Reduction of Bacterial Infection and Inflammation

Puthia,

Petrlova,

Petruk

et al. 2023

Adv Healthcare Materials

Self Cite

View full text Add to dashboard Cite

Surgical site infections (SSI) are a clinical and economic burden. Suture‐associated SSI may develop when bacteria colonize the suture surface and form biofilms that are resistant to antibiotics. Thrombin‐derived C‐terminal peptide (TCP)‐25 is a host defense peptide with a unique dual mode of action that can target both bacteria and the excessive inflammation induced by bacterial products. The peptide demonstrates therapeutic potential in preclinical in vivo wound infection models. In this study, the authors set out to explore whether TCP‐25 can provide a new bioactive innate immune feature to hydrophilic polyglactin sutures (Vicryl). Using a combination of biochemical, biophysical, antibacterial, biofilm, and anti‐inflammatory assays in vitro, in silico molecular modeling studies, along with experimental infection and inflammation models in mice, a proof‐of‐concept that TCP‐25 can provide Vicryl sutures with a previously undisclosed host defense capacity, that enables targeting of bacteria, biofilms, and the accompanying inflammatory response, is shown.

show abstract

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

Cited by 8 publications

References 41 publications

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

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

Label-free high-resolution infrared spectroscopy for spatiotemporal analysis of complex living systems

Bioactive Suture with Added Innate Defense Functionality for the Reduction of Bacterial Infection and Inflammation

Contact Info

Product

Resources

About