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Related Experiment Video

Updated: Apr 20, 2026

A Label-free Technique for the Spatio-temporal Imaging of Single Cell Secretions
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High resolution surface plasmon resonance imaging for single cells.

Alexander W Peterson1, Michael Halter, Alessandro Tona

  • 1Biosystems and Biomaterials Division, National Institute of Standards and Technology, 100 Bureau Drive, Mail Stop 8313, Gaithersburg, MD 20899, USA. alexander.peterson@nist.gov.

BMC Cell Biology
|December 3, 2014
PubMed
Summary
This summary is machine-generated.

Surface plasmon resonance imaging (SPRI) offers label-free visualization of subcellular structures. This technique achieves high spatial resolution, comparable to fluorescence microscopy, without fluorescent labels.

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Real Time Measurements of Membrane Protein:Receptor Interactions Using Surface Plasmon Resonance SPR
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Real Time Measurements of Membrane Protein:Receptor Interactions Using Surface Plasmon Resonance SPR
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Area of Science:

  • Biophysics
  • Cell Biology
  • Microscopy

Background:

  • Surface plasmon resonance imaging (SPRI) is a label-free method for detecting refractive index changes.
  • SPRI has been used to study cell-substratum interactions, but 3D spatial resolution needs characterization.
  • Asymmetric surface plasmon propagation and evanescent wave decay complicate quantitative interpretation of SPRI images.

Purpose of the Study:

  • To experimentally characterize the spatial resolution of an SPR imaging system in lateral and distal dimensions.
  • To demonstrate a novel approach for resolving sub-micrometer cellular structures using SPRI.
  • To provide a basis for quantitative interpretation of SPRI for subcellular imaging.

Main Methods:

  • Designed an SPR imaging system utilizing a high numerical aperture objective lens and a digital light projector for patterned incident light.
  • Visualized cellular components like focal adhesions and the nucleus.
  • Used polymeric nanoparticle beads to determine the point spread function and characterize SPR field detection limits as a function of distance.

Main Results:

  • Achieved near-diffraction limited spatial resolution, as indicated by the point spread function of nanoparticle beads.
  • Characterized the z-axis response using micrometer-scale beads, determining the SPR field's detection limit.
  • Demonstrated multi-wavelength measurements to tailor the evanescent wave's penetration depth into the cellular environment.

Conclusions:

  • Patterned incident light enables SPRI with high spatial resolution and characterized penetration depth.
  • A novel technique provides unprecedented subcellular detail via SPRI.
  • This advancement allows for quantitative interpretation of SPRI in subcellular imaging.