Multifunctional core–shell particles composed of magnetic particles covered with a gold nanoshell can be induced to align into conducting lines upon application of a magnetic field (see Figure). The formation of Au clusters and “streaky” gold nanoparticles on the surface of the PS beads is demonstrated and the preparation, characterization, and applications of magnetic and polystyrene beads featuring a gold shell are addressed.
We developed a novel surface plasmon resonance (SPR) method, based on Fourier transform infrared (FTIR) spectroscopy, as a label-free technique for studying dynamic processes occurring within living cells in real time. With this method, the long (micrometer) infrared wavelength produced by the FTIR generates an evanescent wave that penetrates deep into the sample. In this way, it enables increased depth of sensing changes, covering significant portions of the cell-height volumes. HeLa cells cultivated on a gold-coated prism were subjected to acute cholesterol enrichment or depletion using cyclodextrins. Cholesterol insertion into the cell plasma membrane resulted in an exponential shift of the SPR signal toward longer wavelengths over time, whereas cholesterol depletion caused a shift in the opposite direction. Upon application of the inactive analog alpha-cyclodextrin (alpha-CD), the effects were minimal. A similar trend in the SPR signal shifts was observed on a model membrane system. Our data suggest that FTIR-SPR can be implemented as a sensitive technique for monitoring in real time dynamic changes taking place in living cells.
We discuss the Surface-Plasmon-Resonance (SPR) technique based on Fourier -Transform -In-fraRed (FTIR) spectrometry. We explore the potential of the infrared surface plasmon resonance technique for biological studies in aqueous solutions and compare it to the conventional surface plasmon technique operating in the visible range. We demonstrate that the sensitivity of the SPR technique in the infrared range is not lower and in fact is even higher. We show several examples of applying FTIR-SPR for biological studies: (i) monitoring D-glucose concentration in solution, and (ii) measuring D-glucose uptake by erythrocytes in suspension. We emphasize the advantages of infrared SPR for studying living cell cultures and show how this technique can be used for characterization of (i) cholesterol penetration into plasma membrane, and (ii) transferrin-induced clathrin-mediated endocytosis. * electronic address: golos@vms.huji.ac.il
The development of novel technologies capable of monitoring the dynamics of cell-cell and cell-substrate interactions in real time and a label-free manner is vital for gaining deeper insights into these most fundamental cellular processes. However, the label-free technologies available today provide only limited information on these processes. Here, we report a new (to our knowledge) infrared surface plasmon resonance (SPR)-based methodology that can resolve distinct phases of cell-cell and cell-substrate adhesion of polarized Madin Darby canine kidney epithelial cells. Due to the extended penetration depth of the infrared SP wave, the dynamics of cell adhesion can be detected with high accuracy and high temporal resolution. Analysis of the temporal variation of the SPR reflectivity spectrum revealed the existence of multiple phases in epithelial cell adhesion: initial contact of the cells with the substrate (cell deposition), cell spreading, formation of intercellular contacts, and subsequent generation of cell clusters. The final formation of a continuous cell monolayer could also be sensed. The SPR measurements were validated by optical microscopy imaging. However, in contrast to the SPR method, the optical analyses were laborious and less quantitative, and hence provided only limited information on the dynamics and phases of cell adhesion.
We report on the application of surface plasmon resonance (SPR), based on Fourier transform infrared spectroscopy in the mid-infrared wavelength range, for real-time and label-free sensing of transferrin-induced endocytic processes in human melanoma cells. The evanescent field of the mid-infrared surface plasmon penetrates deep into the cell, allowing highly sensitive SPR measurements of dynamic processes occurring at significant cellular depths. We monitored in real-time, infrared reflectivity spectra in the SPR regime from living cells exposed to human transferrin (Tfn). We show that although fluorescence microscopy measures primarily Tfn accumulation in recycling endosomes located deep in the cell's cytoplasm, the SPR technique measures mainly Tfn-mediated formation of early endocytic organelles located in close proximity to the plasma membrane. Our SPR and fluorescence data are very well described by a kinetic model of Tfn endocytosis, suggested previously in similar cell systems. Hence, our SPR data provide further support to the rather controversial ability of Tfn to stimulate its own endocytosis. Our analysis also yields what we believe is novel information on the role of membrane cholesterol in modulating the kinetics of endocytic vesicle biogenesis and consumption.
We report on a Surface-Plasmon-Resonance (SPR) technique based on Fourier -Transform -Infra -Red (FTIR) spectrometer. In contrast to the conventional surface plasmon technique, operating at a fixed wavelength and a variable angle of incidence, our setup allows the wavelength and the angle of incidence to be varied simultaneously. We explored the potential of the SPR technique in the infrared for biological studies involving aqueous solutions. Using computer simulations, we found the optimal combination of parameters (incident angle, wavelength) for performing this task. Our experiments with physiologically important glucose concentrations in water and in human plasma verified our computer simulations. Importantly, we demonstrated that the sensitivity of the SPR technique in the infrared range is not lower and in fact is even higher than that for visible light.We emphasize the advantages of infra red SPR for studying glucose and other biological molecules in living cells.
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