The Advanced Microscopy & Imaging core facility houses a TCS SP5 multi-photon laser scanning confocal microscope (Leica Microsystems) equipped with both conventional and high-speed resonance scanners. The Multi-photon instrument is attached to an inverted microscope and allows deeper penetration into tissue with less photo-bleaching/damage. Eight laser plus multi-photon excitations are available, producing the following laser excitation lines: 458, 476, 488, 496, 514, 561, 594, 633, and 710-990nm (from the tunable Ti-Sapphire MP laser). The system is capable of collecting up to 5 colors simultaneously for quantitative confocal image analysis, 3D reconstruction, FRAP and FRET, animation, stereo imaging, single layer projection, time lapse collection, and co-localization analysis.
Multi-photon Microscopy using pulsed Infra-Red Laser:

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| Two photons are better than one! A direct comparison of confocal (left) and multi-photon excitation (right) |
Advantages
- No attenuation of exciting light by out of focus fluorescence.
- Longer wavelengths, less scattering and photodamage, deeper penetration
- No scatter of fluorescence light away from focal plane (signal not degraded)
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