• Register
Forgot Password

If you have forgotten your password you can enter your email here and get a temporary password sent to your email.


Leaving Community

Are you sure you want to leave this community? Leaving the community will revoke any permissions you have been granted in this community.


A compact multiphoton 3D imaging system for recording fast neuronal activity.

We constructed a simple and compact imaging system designed specifically for the recording of fast neuronal activity in a 3D volume. The system uses an Yb:KYW femtosecond laser we designed for use with acousto-optic deflection. An integrated two-axis acousto-optic deflector, driven by digitally synthesized signals, can target locations in three dimensions. Data acquisition and the control of scanning are performed by a LeCroy digital oscilloscope. The total cost of construction was one order of magnitude lower than that of a typical Ti:sapphire system. The entire imaging apparatus, including the laser, fits comfortably onto a small rig for electrophysiology. Despite the low cost and simplicity, the convergence of several new technologies allowed us to achieve the following capabilities: i) full-frame acquisition at video rates suitable for patch clamping; ii) random access in under ten microseconds with dwelling ability in the nominal focal plane; iii) three-dimensional random access with the ability to perform fast volume sweeps at kilohertz rates; and iv) fluorescence lifetime imaging. We demonstrate the ability to record action potentials with high temporal resolution using intracellularly loaded potentiometric dye di-2-ANEPEQ. Our design proffers easy integration with electrophysiology and promises a more widespread adoption of functional two-photon imaging as a tool for the study of neuronal activity. The software and firmware we developed is available for download at http://neurospy.org/ under an open source license.

Pubmed ID: 17684546


  • Vucinić D
  • Sejnowski TJ


PloS one

Publication Data

August 8, 2007

Associated Grants

  • Agency: Howard Hughes Medical Institute, Id:

Mesh Terms

  • Animals
  • Electrophysiology
  • Equipment Design
  • Fluorescent Dyes
  • Image Interpretation, Computer-Assisted
  • Imaging, Three-Dimensional
  • Lasers
  • Microscopy, Fluorescence, Multiphoton
  • Neurons
  • Patch-Clamp Techniques
  • Rats
  • Rats, Long-Evans
  • Time Factors
  • Ytterbium