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A Label-free Technique for the Spatio-temporal Imaging of Single Cell Secretions
Published on: November 23, 2015
A new methodology for quantitative LSPR biosensing and imaging
Marc P Raphael1, Joseph A Christodoulides, Shawn P Mulvaney
1Naval Research Laboratory, Washington, DC 20375, USA.
Analytical Chemistry
|January 13, 2012
Summary
This study introduces a new method for localized surface plasmon resonance (LSPR) biosensing to quantify binding events and map them in real-time. The technique precisely measures surface-receptor interactions, offering high temporal and spatial resolution for biosensor applications.
Area of Science:
- Nanotechnology
- Biochemistry
- Surface Science
Background:
- Localized surface plasmon resonance (LSPR) is a powerful optical phenomenon for biosensing.
- Existing LSPR techniques often lack precise quantification of binding kinetics and spatiotemporal mapping capabilities.
Purpose of the Study:
- To develop a quantitative analysis methodology for LSPR biosensing.
- To create an LSPR imaging technique for spatiotemporal mapping of molecular binding events.
- To determine surface-receptor fractional occupancy and binding kinetics.
Main Methods:
- Fabrication of gold nanostructure arrays using electron beam nanolithography.
- Spectroscopic determination of fractional occupancy to measure association kinetics.
- Simultaneous CCD-based LSPR imaging for spatiotemporal mapping of binding events.
- Regeneration of arrays for comparative kinetic analysis across varying analyte concentrations (1 μM to 50 nM).
Main Results:
- Demonstrated a quantitative methodology for LSPR biosensing.
- Achieved spatiotemporal mapping of neutravidin binding to biotinylated surfaces.
- Measured association kinetics with high precision.
- LSPR imaging revealed binding via enhanced scattering with 200 ms temporal and 8 μm² spatial resolution.
Conclusions:
- The developed LSPR methodology enables precise quantification of surface-receptor fractional occupancy.
- The LSPR imaging technique provides effective spatiotemporal mapping of binding events.
- This approach offers a significant advancement for real-time biosensing and kinetic analysis.
