• Corpus ID: 10806984

Two-Photon Fluorescence Microscopy : Basic Principles , Advantages and Risks

  title={Two-Photon Fluorescence Microscopy : Basic Principles , Advantages and Risks},
  author={Sean J. Mulligan and Brian A. MacVicar},
The application of two-photon excitation to fluorescence microscopy has become a powerful tool for studying biological function in live tissue and offers many advantages over conventional imaging techniques. Neuroscientists in particular have used this technology to image physiological functioning in microscopic and subcellular neural compartments. Neurons can be imaged deep within highly light scattering tissue with unparalleled spatial resolution and dramatically reduced photodynamic tissue… 

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Biophys J 77

  • 2226-36
  • 1999

in Confocal and Two-Photon Microscopy; Foundations

  • Applications, and Advances A. Diaspro, Ed.
  • 2002

Annu Rev Biomed Eng 2

  • 399-429
  • 2000

Neuron 18

  • 351-7
  • 2007

Proc Natl Acad Sci U S A 93

  • 10763-8
  • 1996


  • Minta, A Tsien R.Y., J. Am. Chem. Soc. 110, 3212-3220
  • 1988

Biophys J 82

  • 2811-25
  • 2002

Biophys J 75

  • 2015-24
  • 1998