Anatomical and functional imaging of neurons using 2-photon laser scanning microscopy

W. Denk, K. R. Delaney, A. Gelperin, D. Kleinfeld, B. W. Strowbridge, D. W. Tank, R. Yuste

Research output: Contribution to journalArticlepeer-review

290 Scopus citations


Light scattering by brain tissue and phototoxicity are major obstacles to the use of high-resolution optical imaging and photo-activation ('uncaging') of bioactive compounds from inactive ('caged') precursors in intact and semi-intact nervous systems. Optical methods based on 2-photon excitation promise to reduce these obstacles (Denk, 1994; Denk et al., 1990, 1994). Here we show a range of imaging modes based on 2-photon laser scanning microscopy (TPLSM) as applicable to problems in neuroscience. Fluorescence images were taken of neurons labeled with ion-sensitive and voltage-sensitive dyes in invertebrate ganglia, mammalian brain slices, and from the intact mammalian brain. Scanning photochemical images with whole-cell current detection (Denk, 1994) show how the distribution of neurotransmitter receptors on the surface of specific cells can be mapped. All images show strong optical sectioning and usable images can be obtained at depths greater than 100 μm below the surface of the preparation.

Original languageEnglish (US)
Pages (from-to)151-162
Number of pages12
JournalJournal of Neuroscience Methods
Issue number2
StatePublished - Oct 1994

All Science Journal Classification (ASJC) codes

  • General Neuroscience


  • Light scattering
  • Microscopy, 2-photon laser scanning
  • Phototoxicity
  • Receptor, neurotransmitter


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