Simultaneous two-photon imaging and photo-stimulation with structured light illumination

Opt Express. 2010 Aug 30;18(18):18720-31. doi: 10.1364/OE.18.018720.


Holographic microscopy is increasingly recognized as a promising tool for the study of the central nervous system. Here we present a "holographic module", a simple optical path that can be combined with commercial scanheads for simultaneous imaging and uncaging with structured two-photon light. The present microscope is coupled to two independently tunable lasers and has two principal configurations: holographic imaging combined with galvo-steered uncaging and holographic uncaging combined with conventional scanning imaging. We applied this flexible system for simultaneous two-photon imaging and photostimulation of neuronal cells with complex light patterns, opening new perspectives for the study of brain function in situ and in vivo.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Brain / pathology*
  • Diagnostic Imaging / methods*
  • Equipment Design
  • Light
  • Mice
  • Mice, Inbred C57BL
  • Microscopy, Fluorescence / methods*
  • Neurons / pathology*
  • Optics and Photonics
  • Photons
  • Polylysine / chemistry
  • Silicon / chemistry
  • Software


  • Polylysine
  • Silicon