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Leica SP8 Two-Photon
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Zeiss LSM880 Confocal
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How to get started at the AOMF

O-MUG

Comprehensive Course on Fluorescence Microscopy


Leica SP8 Two-Photon/Confocal


In two-photon microscopy, a pulsed infra-red laser scans across the sample to produce a confocal-like (i.e., optically sectioned) image of a fluorescent specimen. Two-photon microscopy can have several advantages over confocal microscopy for thick specimens, including deeper penetration and reduced photobleaching outside of the focal plane.

On our system, we have the newest Coherent Discovery laser for two-photon excitation that can be tuned in the 680-1300 nm range. The system is also a fully-functional confocal microscope. There are 2 "normal" (single-photon) lasers that together provide the following excitation lines: 488 and 555 nm. The system is built on an upright platform, with single objective lens and adjustable/motorized stage, making it ideal for intravital imaging. There are also two pairs of non-descanned detectors that can be used to achieve increased sensitivity. These include 2 PMT and 2 GaAsP detectors. This is also the "resonant" SP8, meaning that in addition to the standard galvanometric scan mode (~ 1 fps), a high speed resonant scanning mode (30+ fps) is also possible.

Training Courses Available
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Confocal/Multi-photon Microscope Training
Advanced Confocal/Multi-photon Techniques

Model
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Leica TCS SP8 MP

Objective Lens
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HC IRAPO L 25x/1.0 W motCORR
Motorized collar for reproducible spherical aberration correction.

Illumination Sources
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Diode 488nm
Diode 555nm
Coherent Discovery femtosecond laser for two-photon (680-1300nm)

Advantages and Techniques
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Intravital imaging
High speed confocal imaging
High resolution confocal and two-photon fluorescence imaging
Optical sectioning of thick fluorescent specimens
Deep penetration into thick specimens as compared to Confocal
Less photobleaching outside of the focal plane as compared to Confocal
Spectral unmixing


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