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) |
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Pages (from-to) | 398-408 |
Number of pages | 11 |
Journal | Pflugers Archiv European Journal of Physiology |
Volume | 441 |
Issue number | 2-3 |
DOIs | |
State | Published - Dec 1 2000 |
Externally published | Yes |
Keywords
- Fluoview
- Laser
- Pockels
- Scanning
- Spines
- XCOSM
ASJC Scopus subject areas
- Physiology
- Clinical Biochemistry
- Physiology (medical)