Abstract
Confocal fluorescence microscopy coupled with a diffraction-limited laser beam and a high-efficiency detection system has been used to study the diffusive movement and emission process of individual fluorescent molecules in the liquid phase at room temperature. The high detection sensitivity achieved at fast data acquisition speeds (greater than 1 kilohertz) allows real-time observation of single-molecule fluorescence without statistical analysis. The results show fluorescence-cycle saturation at the single-molecule level and multiple recrossings of a single molecule into and out of the probe volume as well as the triplet state.
Keywords
FluorescenceConfocalLaser-induced fluorescenceConfocal microscopyMicroscopyMicroscopeFluorescence in the life sciencesMoleculeFluorescence microscopeFluorescence cross-correlation spectroscopyMaterials scienceFluorescence-lifetime imaging microscopyAnalytical Chemistry (journal)ChemistryOpticsPhysicsChromatography
Affiliated Institutions
Related Publications
Imaging of single molecule diffusion.
In recent years observations at the level of individual atoms and molecules became possible by microscopy and spectroscopy. Imaging of single fluorescence molecules has been ach...
Alterations of Single Molecule Fluorescence Lifetimes in Near-Field Optical Microscopy
Fluorescence lifetimes of single Rhodamine 6G molecules on silica surfaces were measured with pulsed laser excitation, time-correlated single photon counting, and near-field sca...
Single-Molecule Fluorescence Analysis in Solution
Over the past five years, several groups have developed the capability to detect and identify single fluorescent molecules in solution as the molecules flow through a focused la...
Imaging and Time-Resolved Spectroscopy of Single Molecules at an Interface
Far-field microscopy was used to noninvasively measure the room-temperature optical properties of single dye molecules located on a polymer-air interface. Shifts in the fluoresc...
Single Molecules Observed by Near-Field Scanning Optical Microscopy
Individual carbocyanine dye molecules in a sub-monolayer spread have been imaged with near-field scanning optical microscopy. Molecules can be repeatedly detected and spatially ...
Publication Info
- Year
- 1994
- Type
- article
- Volume
- 266
- Issue
- 5187
- Pages
- 1018-1021
- Citations
- 545
- Access
- Closed
External Links
Social Impact
Altmetric
PlumX Metrics
Social media, news, blog, policy document mentions
Citation Metrics
545
OpenAlex
Cite This
Shuming Nie,
Daniel T. Chiu,
Richard N. Zare
(1994).
Probing Individual Molecules with Confocal Fluorescence Microscopy.
Science
, 266
(5187)
, 1018-1021.
https://doi.org/10.1126/science.7973650
Identifiers
- DOI
- 10.1126/science.7973650