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Atomic Force Microscopy01:08

Atomic Force Microscopy

Atomic force microscopy (AFM) is a type of scanning probe microscopy that can analyze topographic details of various specimens like ceramics, glass, polymers, and biological samples. AFM offers over 1000 times more resolution than the optical imaging system. Images generated from AFM are three-dimensional surface profiles, offering an advantage over the flat, two-dimensional images from other imaging techniques.
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Related Experiment Video

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Functionalization of Atomic Force Microscope Cantilevers with Single-T Cells or Single-Particle for Immunological Single-Cell Force Spectroscopy
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QD as a bifunctional cell-surface marker for both fluorescence and atomic force microscopy.

Yunqi Wang1, Yong Chen, Jiye Cai

  • 1Department of Microbiology & Immunology, University of Illinois at Chicago, Chicago, IL 60612, USA. yqwang@uic.edu

Ultramicroscopy
|January 24, 2009
PubMed
Summary

Fluorescent quantum dots (QDs) offer enhanced cell imaging by labeling cell-surface receptors. This study shows QDs can simultaneously provide fluorescence and atomic force microscopy images of human red blood cells.

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Fluorescence Biomembrane Force Probe: Concurrent Quantitation of Receptor-ligand Kinetics and Binding-induced Intracellular Signaling on a Single Cell

Published on: August 4, 2015

Area of Science:

  • Biophysics
  • Nanotechnology
  • Cell Biology

Background:

  • Fluorescent quantum dots (QDs) are advanced fluorescent labels utilized in cell imaging.
  • Streptavidin-conjugated QDs (10-15 nm) enhance contrast in atomic force microscopy (AFM) for biomolecule labeling.
  • QD properties enable correlative fluorescence microscopy (FM) and AFM imaging.

Purpose of the Study:

  • To assess QD-streptavidin conjugates as probes for labeling wheat germ agglutinin (WGA) receptors on human red blood cells (RBCs).
  • To evaluate the feasibility of simultaneous fluorescence microscopy (FM) and AFM imaging using QD probes.
  • To investigate the distribution and expression of membrane proteins and receptors on cell surfaces.

Main Methods:

  • Utilized QD-streptavidin conjugates to label WGA receptors on human RBC membranes.
  • Acquired simultaneous fluorescence and AFM images of labeled RBCs.
  • Analyzed QD distribution and density on individual erythrocytes.

Main Results:

  • QD labeling on human RBCs exhibited non-uniform distribution.
  • Significant variations in the number of labeled QDs were observed across different erythrocytes.
  • QD distribution patterns may correlate with erythrocyte age.

Conclusions:

  • QD-streptavidin conjugates serve as effective bifunctional probes for simultaneous FM and AFM.
  • QDs facilitate quantitative investigation of membrane protein and receptor distribution and expression.
  • This technique offers a novel approach for analyzing cell surface heterogeneity.