Surface plasmon excitation using a Fourier-transform infrared spectrometer: Live cell and bacteria sensing

Lirtsman, Vladislav; Golosovsky, M.; Davidov, D.

doi:10.1063/1.4997388

Cited by 7 publications

(3 citation statements)

References 39 publications

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“…Therefore, an important way would be to develop biosensors with greater penetration depth of the evanescent field, to possibly detect NP admission for the lower concentrations of NPs as well. The adaptation of deep-probing sensors − into high-throughput formats would find interesting applications in the present field.…”

Section: Discussionmentioning

confidence: 99%

Interaction of Positively Charged Gold Nanoparticles with Cancer Cells Monitored by an in Situ Label-Free Optical Biosensor and Transmission Electron Microscopy

Péter

Lagzi

Teraji

et al. 2018

ACS Appl. Mater. Interfaces

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Functionalized nanoparticles (NPs) can penetrate into living cells and vesicles, opening up an extensive range of novel directions. For example, NPs are intensively employed in targeted drug delivery and biomedical imaging. However, the real-time kinetics and dynamics of NP-living cell interactions remained uncovered. In this study, we in situ monitored the cellular uptake of gold NPs-functionalized with positively charged alkaline thiol-into surface-adhered cancer cells, by using a high-throughput label-free optical biosensor employing resonant waveguide gratings. The characteristic kinetic curves upon NP exposure of cell-coated biosensor surfaces were recorded and compared to the kinetics of NP adsorption onto bare sensor surfaces. We demonstrated that from the above kinetic information, one can conclude about the interactions between the living cells and the NPs. Real-time biosensor data suggested the cellular uptake of the functionalized NPs by an active process. It was found that positively charged particles penetrate into the cells more effectively than negatively charged control particles, and the optimal size for the cellular uptake of the positively charged particles is around 5 nm. These conclusions were obtained in a cost-effective, fast, and high-throughput manner. The fate of the NPs was further revealed by electron microscopy on NP-exposed and subsequently fixed cells, well confirming the results obtained by the biosensor. Moreover, an ultrastructural study demonstrated the involvement of the endosomal-lysosomal system in the uptake of functionalized NPs and suggested the type of the internalization pathway.

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Section: Discussionmentioning

confidence: 99%

Interaction of Positively Charged Gold Nanoparticles with Cancer Cells Monitored by an in Situ Label-Free Optical Biosensor and Transmission Electron Microscopy

Péter

Lagzi

Teraji

et al. 2018

ACS Appl. Mater. Interfaces

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“…Two approaches have been used to achieve a greater penetration depth of the plasmon wave: excitation of long-range surface plasmon in a specific SPR biochip [5] and plasmon excitation in the UV range [6,7]. The latter approach seems to be more effective-the penetration depth could reach several microns.…”

Section: Penetration Depth and Detection Depthmentioning

confidence: 99%

SPR-Based Kinetic Analysis of the Early Stages of Infection in Cells Infected with Human Coronavirus and Treated with Hydroxychloroquine

et al. 2021

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Cell-based assays are a valuable tool for examination of virus–host cell interactions and drug discovery processes, allowing for a more physiological setting compared to biochemical assays. Despite the fact that cell-based SPR assays are label-free and thus provide all the associated benefits, they have never been used to study viral growth kinetics and to predict drug antiviral response in cells. In this study, we prove the concept that the cell-based SPR assay can be applied in the kinetic analysis of the early stages of viral infection of cells and the antiviral drug activity in the infected cells. For this purpose, cells immobilized on the SPR slides were infected with human coronavirus HCov-229E and treated with hydroxychloroquine. The SPR response was measured at different time intervals within the early stages of infection. Methyl Thiazolyl Tetrazolium (MTT) assay was used to provide the reference data. We found that the results of the SPR and MTT assays were consistent, and SPR is a reliable tool in investigating virus–host cell interaction and the mechanism of action of viral inhibitors. SPR assay was more sensitive and accurate in the first hours of infection within the first replication cycle, whereas the MTT assay was not so effective. After the second replication cycle, noise was generated by the destruction of the cell layer and by the remnants of dead cells, and masks useful SPR signals.

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“…In the last few decades, label-free optical biosensors have proven their unique capabilities in a wide range of applications from biomolecular interaction analysis (BIA) [1][2][3][4] to whole-cell monitoring [5][6][7][8][9] . Screening the interactions of various analyte-ligand pairs has become essential in drug discovery [10][11][12][13] , assay and sensor development 14,15 as well as for understanding molecular mechanisms in biochemistry and biophysics [16][17][18][19] .…”

mentioning

confidence: 99%

Grating-coupled interferometry reveals binding kinetics and affinities of Ni ions to genetically engineered protein layers

Jankovics

Kovács

Saftics

et al. 2020

Sci Rep

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Reliable measurement of the binding kinetics of low molecular weight analytes to their targets is still a challenging task. Often, the introduction of labels is simply impossible in such measurements, and the application of label-free methods is the only reliable choice. By measuring the binding kinetics of Ni(II) ions to genetically modified flagellin layers, we demonstrate that: (1) Grating-Coupled Interferometry (GCI) is well suited to resolve the binding of ions, even at very low protein immobilization levels; (2) it supplies high quality kinetic data from which the number and strength of available binding sites can be determined, and (3) the rate constants of the binding events can also be obtained with high accuracy. Experiments were performed using a flagellin variant incorporating the C-terminal domain of the nickel-responsive transcription factor NikR. GCI results were compared to affinity data from titration calorimetry. We found that besides the low-affinity binding sites characterized by a micromolar dissociation constant (Kd), tetrameric FliC-NikRC molecules possess high-affinity binding sites with Kd values in the nanomolar range. GCI enabled us to obtain real-time kinetic data for the specific binding of an analyte with molar mass as low as 59 Da, even at signals lower than 1 pg/mm2.

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Surface plasmon excitation using a Fourier-transform infrared spectrometer: Live cell and bacteria sensing

Cited by 7 publications

References 39 publications

Interaction of Positively Charged Gold Nanoparticles with Cancer Cells Monitored by an in Situ Label-Free Optical Biosensor and Transmission Electron Microscopy

Interaction of Positively Charged Gold Nanoparticles with Cancer Cells Monitored by an in Situ Label-Free Optical Biosensor and Transmission Electron Microscopy

SPR-Based Kinetic Analysis of the Early Stages of Infection in Cells Infected with Human Coronavirus and Treated with Hydroxychloroquine

Grating-coupled interferometry reveals binding kinetics and affinities of Ni ions to genetically engineered protein layers

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