Abstract
A new method for in vitro and possibly in vivo ultrahigh-resolution colocalization and distance measurement between biomolecules is described, based on semiconductor nanocrystal probes. This ruler bridges the gap between FRET and far-field (or near-field scanning optical microscope) imaging and has a dynamic range from few nanometers to tens of micrometers. The ruler is based on a stage-scanning confocal microscope that allows the simultaneous excitation and localization of the excitation point-spreadfunction (PSF) of various colors nanocrystals while maintaining perfect registry between the channels. Fit of the observed diffraction and photophysics-limited images of the PSFs with a two-dimensional Gaussian allows one to determine their position with nanometer accuracy. This new high-resolution tool opens new windows in various molecular, cell biology and biotechnology applications.
| Original language | English |
|---|---|
| Pages (from-to) | 8-15 |
| Number of pages | 8 |
| Journal | Proceedings of SPIE - The International Society for Optical Engineering |
| Volume | 4258 |
| DOIs | |
| State | Published - Jan 2001 |
| Externally published | Yes |
| Event | Nanoparticles and Nanostrutured Surfaces: Novel Reporters with Biological Applications - San Jose, CA, United States Duration: 24 Jan 2001 → 25 Jan 2001 |
Keywords
- Confocal
- Diffraction limit
- Fluorescence
- Microscopy
- Quantum dot
- Semiconductor nanocrystal
- Single molecule
- Superresolution