Photon Upmanship: Why Multiphoton Imaging Is More than a Gimmick

@article{Denk1997PhotonUW,
  title={Photon Upmanship: Why Multiphoton Imaging Is More than a Gimmick},
  author={W. Denk and K. Svoboda},
  journal={Neuron},
  year={1997},
  volume={18},
  pages={351-357}
}
The unique niche that light microscopy occupies in biology is based on the ability to perform observations on living tissue at relatively high spatial resolution. This resolution is limited by the wavelength of light and does not rival that of electron microscopy, which is, however, a fundamentally nonvital form of observation. Other vital microscopies, such as MRI, can neither resolve subcellular structures nor provide the exquisite molecular selectivity that allows the detection of even… Expand
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References

SHOWING 1-10 OF 60 REFERENCES
Intravital imaging of green fluorescent protein using two-photon laser-scanning microscopy.
TLDR
It is shown that TPLSM provides greater sensitivity, better resolution and less photo-bleaching, as compared to confocal laser-scanning microscopy, which allows a safer and higher-resolution means of imaging living cells labeled with a variety of fluorophores, including green fluorescent protein. Expand
NONLINEAR ABSORPTION EXTENDS CONFOCAL FLUORESCENCE MICROSCOPY INTO THE ULTRA-VIOLET REGIME AND CONFINES THE ILLUMINATION VOLUME
It is shown that two-photon absorption confines the illumination volume and present quantitative evidence that an additional confocal arrangement of the detector further improves the resolution by… Expand
Time-resolved fluorescence imaging and background rejection by two-photon excitation in laser-scanning microscopy
A new dimension in quantitative fluorescence microscopy may be accessed by imaging of fluorescence decay times. To obtain spatially resolved information from microscopic sample locations, one must… Expand
Two-photon excitation in functional biological imaging.
  • W. Denk
  • Physics, Medicine
  • Journal of biomedical optics
  • 1996
TLDR
The contribution that the combination of two-photon absorption with laser scanning20 has made to functional biological imaging in recent years is reviewed. Expand
Multiphoton fluorescence excitation: new spectral windows for biological nonlinear microscopy.
TLDR
The measured nonlinear excitation spectra and their photophysical characteristics that empower nonlinear laser microscopy for biological imaging are described. Expand
Anatomical and functional imaging of neurons using 2-photon laser scanning microscopy
TLDR
A range of imaging modes based on 2-photon laser scanning microscopy (TPLSM) are shown as applicable to problems in neuroscience, showing how the distribution of neurotransmitter receptors on the surface of specific cells can be mapped. Expand
Use of confocal imaging in the study of biological structures.
TLDR
This work has developed a high-speed beam scanning confocal imaging system that can be used in conjunction with a conventional microscope, and has examined a variety of biological material using this system, finding that confocal Imaging gives a marked improvement in quality over conventional techniques. Expand
Measuring Serotonin Distribution in Live Cells with Three-Photon Excitation
TLDR
Three-dimensionally resolved images are presented along with measurements of the serotonin concentration and content and content of individual secretory granules from tryptophan and monoamines serotonin and dopamine. Expand
Two-photon laser scanning fluorescence microscopy.
TLDR
The fluorescence emission increased quadratically with the excitation intensity so that fluorescence and photo-bleaching were confined to the vicinity of the focal plane as expected for cooperative two-photon excitation. Expand
Application of a femtosecond self-sustaining mode-locked Ti:sapphire laser to the field of laser scanning confocal microscopy
Developments in ultrafast Ti:sapphire laser technology can be applied in the investigation of nonlinear optical processes. We describe the application of a self-sustaining femtosecond Ti:sapphire… Expand
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