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  • Overcoming the hurdle of single protein analysis by Plasmonic Scattering Microscopy

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Overcoming the hurdle of single protein analysis by Plasmonic Scattering Microscopy
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Overcoming the hurdle of single protein analysis by Plasmonic Scattering Microscopy

bioxone September 24, 2020September 23, 2020

Sristi Raj Rai, Amity University Kolkata

Free electrons of limited elements (Au, Ag, Cu) has the optical property to hover over surfaces when excited, forming an electron cloud. When an electromagnetic wave strikes such electrically conducting metal interfaces, the collective and rapid oscillation of electrons gives rise to the conversion of photons into plasmons. The propagating wave of plasmons can be detected either by inelastic scattering of charged particle or resonant scattering of light. SPR microscopy (SPRM) applies the phenomenon of surface plasmon resonance (SPR) to allow sensitive imaging of biological samples with high spatial resolution.

Protein, the building block of life forms, physically interacts with various molecules in the dynamic system, leading to specific binding via numerous bond formations. These ligand-protein interactions reach a chemical equilibrium influencing the kinetics of the system. Estimation and interpretation of protein binding kinetics of single proteins was a crucial issue in the field of protein analysis due to its even smaller size than virions by SPRM.

In the evolving field of optical biosensing, the label-free real-time detection of SPR scattering from the top of the sensor was employed for imaging single proteins by Pengfei Zhang et al. for the first time. Allowing us to measure the size and identify single proteins based on their specific binding to antibodies. They also provided a digital method to analyze the intrinsic properties of individual molecules, study and quantify molecular protein binding kinetics, and heterogeneity of their behavior. Therefore, plasmonic scattering microscopy (PSM) is an exceptional tool in our toolbox to inspect low volume samples from single cells in the field of biotechnology and medical sciences.

SOURCE – Zhang, P., Ma, G., Dong, W. et al. Plasmonic scattering imaging of single proteins and binding kinetics. Nat Methods (2020). https://doi.org/10.1038/s41592-020-0947-0

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Tagged Analytical biochemistry Plasmonic scattering microscopy (PSM) Protein binding kinetics Scattering Single protein analysis Single-molecule biophysics SPR imaging (SPRi) SPR microscopy (SPRM) Surface plasmon resonance (SPR)

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