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Related Concept Videos

Super-resolution Fluorescence Microscopy01:37

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Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been...
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Related Experiment Video

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Conducting Multiple Imaging Modes with One Fluorescence Microscope
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Towards single molecule biosensors using super-resolution fluorescence microscopy.

Xun Lu1, Philip R Nicovich2, Katharina Gaus2

  • 1School of Chemistry, Australian Centre for NanoMedicine and the ARC Centre of Excellence in Convergent Bio-Nano Science and Technology, University of New South Wales, Sydney 2052, Australia.

Biosensors & Bioelectronics
|November 11, 2016
PubMed
Summary
This summary is machine-generated.

This study shows single molecule localisation microscopy (SMLM) can monitor antibody-antigen binding events on surfaces. This technique offers a new way to characterize biosensing interfaces for quantitative analysis.

Keywords:
ImmunosensorsIndium tin oxideSelf-assembled monolayersSingle molecule localisation microscopySuperresolution fluorescence microscopy

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Area of Science:

  • Biosensing and Interface Science
  • Single-molecule Biophysics
  • Surface Chemistry

Background:

  • Conventional immunosensors rely on bulk measurements, lacking molecular-level characterization of interfaces.
  • Designing immunosensing interfaces requires precise control over antibody immobilization and nonspecific adsorption.
  • Current methods lack tools to assess the molecular-level performance of these sophisticated interfaces.

Purpose of the Study:

  • To investigate the feasibility of using single molecule localisation microscopy (SMLM) for monitoring antibody-antigen binding events.
  • To establish indium tin oxide (ITO) surfaces as a viable platform for SMLM in biosensing applications.
  • To demonstrate the potential of SMLM for characterizing biosensing interfaces at the single-molecule level.

Main Methods:

  • Utilized indium tin oxide (ITO) surfaces for SMLM.
  • Modified ITO surfaces with self-assembled monolayers using organophosphonic acid derivatives.
  • Controlled and monitored antigen and antibody immobilization at the single-molecule level.
  • Observed antibody binding to antigen-modified surfaces using SMLM.

Main Results:

  • Confirmed ITO surfaces are suitable for SMLM.
  • Demonstrated control over antigen and antibody density on the surface.
  • Showed that antibody binding is dependent on both surface antigen concentration and solution antibody concentration.
  • Validated SMLM's capability to monitor binding events at the single-molecule level.

Conclusions:

  • Single molecule localisation microscopy (SMLM) can effectively monitor antibody-antigen binding events.
  • SMLM provides a powerful tool for characterizing biosensing interfaces at the molecular level.
  • This approach paves the way for massively parallel, single-molecule detection schemes for quantitative biosensing.