Pflügers Archiv

, Volume 441, Issue 2, pp 398–408

A custom-made two-photon microscope and deconvolution system

  • Ania Majewska
  • Glen Yiu
  • Rafael Yuste
Instruments and Techniques

DOI: 10.1007/s004240000435

Cite this article as:
Majewska, A., Yiu, G. & Yuste, R. Pflügers Arch - Eur J Physiol (2000) 441: 398. doi:10.1007/s004240000435

Abstract.

We describe in detail a custom-built two-photon microscope based on a modified confocal scanhead (Olympus Fluoview) and mode-locked Ti:sapphire laser (Coherent Mira 900). This system has internal detectors as well as external whole-field detection and an electro-optical modulator for blanking the beam on flyback and effecting fast changes in excitation intensity. This microscope can be used in deep, scattering samples for quantitative measurements with a wide range of fluorophores (GFP, fura, calcium green, calcium orange, fluo-3, DiI, DiO, fluorescein, rhodamine), for fluorescent photobleaching recovery and for uncaging. Images obtained with this system can be deconvolved with the Estimation Maximization algorithm using the program XCOSM (freeware available at: http://www.ibc.wustl.edu/bcl/xcosm/).

Fluoview Laser Pockels Scanning Spines XCOSM 

Copyright information

© Springer-Verlag 2000

Authors and Affiliations

  • Ania Majewska
    • 1
  • Glen Yiu
    • 1
  • Rafael Yuste
    • 1
  1. 1.Department of Biological Sciences, Columbia University, 1212 Amsterdam Avenue, Box 2435, New York, NY 10027, USA