Two-photon laser scanning fluorescence microscopy.

  title={Two-photon laser scanning fluorescence microscopy.},
  author={Winfried Denk and James H. Strickler and Watt W. Webb},
  volume={248 4951},
Molecular excitation by the simultaneous absorption of two photons provides intrinsic three-dimensional resolution in laser scanning fluorescence microscopy. The excitation of fluorophores having single-photon absorption in the ultraviolet with a stream of strongly focused subpicosecond pulses of red laser light has made possible fluorescence images of living cells and other microscopic objects. The fluorescence emission increased quadratically with the excitation intensity so that fluorescence… Expand
Development of a confocal laser scanning fluorescence microscope using two-photon excitation in combination with time-gated detection
Fluorescent molecules having single-photon absorption in the blue and the UV can be excited with infra-red light via a process known as two-photon excitation. The combination of this technique withExpand
Three-photon excitation in fluorescence microscopy.
Three-photon excitation fluorescence axial images are shown of polystyrene beads stained with the fluorophore 2,5-bis(4-biphenyl)oxazole (BBO) using a mode-locked titanium sapphire laser, proving the feasibility of scanning fluorescence microscopy by three-ph photon excitation. Expand
Three-photon light-sheet fluorescence microscopy.
The first demonstration of three-photon excitation light-sheet fluorescence microscopy is presented, using a conventional femtosecond pulsed laser at 1000 nm wavelength for the imaging of 450 μm diameter cellular spheroids. Expand
Two-Photon Microscopy in Highly Scattering Tissue
Molecular excitation by two-photon absorption and the subsequent fluorescence have proved to be a useful tool for imaging biological systems using laser-scanning microscopy (Denk et al., 1990). InExpand
Combined Raman and continuous-wave-excited two-photon fluorescence cell imaging.
With this microscope fast image acquisition with fluorescence imaging can be used to select areas of interest for subsequent chemical analysis with spontaneous Raman imaging. Expand
Two-photon excitation fluorescence microscopy.
Two-photon fluorescence microscopy is one of the most important recent inventions in biological imaging and is a novel method to trigger localized photochemical reactions. Expand
Multi-Photon Fluorescence Microscopy
Multi-photon microscopy uses a different principle in exciting fluorochromes from conventional epi-fluorescence microscopy and confocal laser scanning fluorescence microscopy. In multi-photonExpand
Two-photon excitation in scanning laser microscopy (Abstract Only)
Two-photon excitation in scanning laser fluorescence microscopy provides axial resolution and rejection of out of focus background similar to that provided by confocal microscopy. Two- photonExpand
Combined scanning optical coherence and two-photon-excited fluorescence microscopy.
Simultaneous imaging of cell nuclei with OCM and TPE is demonstrated in live drosophila embryos and both modes provide rapid en-face imaging with submicrometer resolution. Expand
Visible-wavelength two-photon excitation microscopy for fluorescent protein imaging
The calculation of point spread functions shows that the visible-wavelength two-photon excitation provides the fundamental improvement of spatial resolution compared to conventional confocal microscopy. Expand


An evaluation of confocal versus conventional imaging of biological structures by fluorescence light microscopy
It is found that confocal imaging gives greatly enhanced images of biological structures viewed with epifluorescence, and the improvements are such that it is possible to optically section thick specimens with little degradation in the image quality of interior sections. Expand
Two-photon absorption of 7-hydroxycoumarine
Abstract The two-photon absorption spectra of 7-hydroxycoumarine (umbelliferone) in ethanol have been studied in the region 510–650 nm by monitoring the fluorescence excited by a pulsed tunable dyeExpand
Simultaneous formation of solitons and dispersive waves in a femtosecond ring dye laser.
We report the experimental observation of soliton and dispersive pulses generated simultaneously in a passively mode-locked dye laser. The results are interpreted by analogy to the predictions of theExpand
Synthesis, photochemistry, and biological activity of a caged photolabile acetylcholine receptor ligand.
The approach described may be useful in the preparation and characterization of other photolabile precursors of neurotransmitters that contain amino groups, and both properties were found to depend on the nature of the substituents on the photolABile protecting group. Expand
Design considerations for a femtosecond pulse laser balancing self phase modulation, group velocity dispersion, saturable absorption, and saturable gain
This paper describes the design and operating characteristics of a femtosecond pulse ring dye laser that employs the combined effects of self phase modulation, group velocity dispersion, saturableExpand
Three-Dimensional Optical Storage Memory
A novel three-dimensional optical memory device is presented that allows fast random access of the information and extremely high bit densities and Absorption and emission data show that two-photon writing and reading of information is feasible. Expand
The Department of Biochemistry is internationally recognized for its research and education and offers a world-class interdisciplinary research environment in a beautiful mountain setting. As part ofExpand
Two - photon laser microscopy
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