Fluorescence microscopy with diffraction resolution barrier broken by stimulated emission
AUTOR(ES)
Klar, Thomas A.
FONTE
The National Academy of Sciences
RESUMO
The diffraction barrier responsible for a finite focal spot size and limited resolution in far-field fluorescence microscopy has been fundamentally broken. This is accomplished by quenching excited organic molecules at the rim of the focal spot through stimulated emission. Along the optic axis, the spot size was reduced by up to 6 times beyond the diffraction barrier. The simultaneous 2-fold improvement in the radial direction rendered a nearly spherical fluorescence spot with a diameter of 90–110 nm. The spot volume of down to 0.67 attoliters is 18 times smaller than that of confocal microscopy, thus making our results also relevant to three-dimensional photochemistry and single molecule spectroscopy. Images of live cells reveal greater details.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=26924Documentos Relacionados
- Nonlinear structured-illumination microscopy: Wide-field fluorescence imaging with theoretically unlimited resolution
- Shattering the diffraction limit of light: A revolution in fluorescence microscopy?
- Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation
- High-speed, random-access fluorescence microscopy: I. High-resolution optical recording with voltage-sensitive dyes and ion indicators.
- Direct observation of iron-induced conformational changes of mitochondrial DNA by high-resolution field-emission in-lens scanning electron microscopy.