A custom-made two-photon microscope and deconvolution system

Ania Majewska, Glen Yiu, Rafael Yuste

Research output: Contribution to journalArticle

143 Scopus citations

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 photo-bleaching 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/).

Original languageEnglish (US)
Pages (from-to)398-408
Number of pages11
JournalPflugers Archiv European Journal of Physiology
Volume441
Issue number2-3
DOIs
StatePublished - Dec 1 2000
Externally publishedYes

Keywords

  • Fluoview
  • Laser
  • Pockels
  • Scanning
  • Spines
  • XCOSM

ASJC Scopus subject areas

  • Physiology
  • Clinical Biochemistry
  • Physiology (medical)

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