A custom-made two-photon microscope and deconvolution system

Ania Majewska; Glen Yiu; Rafael Yuste

doi:10.1007/s004240000435

A custom-made two-photon microscope and deconvolution system

Ania Majewska, Glen Yiu, Rafael Yuste

Research output: Contribution to journal › Article › peer-review

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 language	English (US)
Pages (from-to)	398-408
Number of pages	11
Journal	Pflugers Archiv European Journal of Physiology
Volume	441
Issue number	2-3
DOIs	https://doi.org/10.1007/s004240000435
State	Published - Dec 1 2000
Externally published	Yes

Keywords

Fluoview
Laser
Pockels
Scanning
Spines
XCOSM

ASJC Scopus subject areas

Physiology
Clinical Biochemistry
Physiology (medical)

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10.1007/s004240000435

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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/).",

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