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Near-field scanning optical microscopy in liquid for high resolution single molecule detection on dendritic cells
M Koopman1, A Cambi, B I de Bakker
1Applied Optics group, Faculty of Science and Technology and MESA+ Institute for Nanotechnology, University of Twente, P.O. Box 217, 7500 AE Enschede, The Netherlands. m.koopman@utwente.nl
FEBS Letters
|August 26, 2004
Summary
Near-field scanning optical microscopy (NSOM) now visualizes cell surface domains in liquid. This breakthrough allows imaging of nanoscale domains on dendritic cells with high resolution.
Area of Science:
- Cell Biology
- Microscopy
- Biophysics
Background:
- Cell surface receptor clustering into micro-domains regulates cellular functions.
- These domains are typically too small for conventional optical microscopy.
- Near-field scanning optical microscopy (NSOM) offers high resolution (down to 70 nm) and single-molecule sensitivity.
Purpose of the Study:
- To extend NSOM capabilities for imaging in liquid environments.
- To visualize nanoscale domains on cell surfaces under physiological conditions.
- To investigate the distribution of DC-SIGN on dendritic cells.
Main Methods:
- Development of a 'diving bell' concept for NSOM operation in liquid.
- Imaging individual fluorescent molecules on cell membranes in solution.
- Utilizing NSOM with 90 nm spatial resolution.
Main Results:
- Successful operation of NSOM in liquid environments.
- First-time imaging of individual fluorescent molecules on cell membranes in solution.
- Direct visualization of nanometric domains of DC-SIGN on dendritic cells in both air and liquid.
Conclusions:
- The 'diving bell' NSOM technique enables high-resolution imaging in liquid.
- This method allows direct visualization of nanoscale cell surface domains.
- The distribution of DC-SIGN on dendritic cells can now be studied in their native environment.