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How to get started at the AOMF

O-MUG

Comprehensive Course on Fluorescence Microscopy


Zeiss LSM710 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 a Chameleon laser for two-photon excitation that can be tuned in the 705-980nm range. The system is also a fully-loaded confocal microscope. There are 4 "normal" (single-photon) lasers that together provide 6 excitation lines from the blue to red: 405, 458, 488, 514, 561, and 633nm. The system is built on an upright platform, with special objective lenses and adjustable stage and nosepiece heights, making it ideal for intravital imaging. There are also a pair of non-descanned Gasp detectors that can be used to achieve increased sensitivity.

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

Model
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Zeiss LSM 710 NLO

Objective Lenses
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5x/0.25 NA
Plan-Apo 10x.0.45 NA
20x/1.0 NA (coverslip corrected)
C-Apo 63x/1.4 NA(oil)

Illumination Sources
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Diode 405nm
Argon 458, 488, 514nm
DPSS 561nm
HeNe 633nm
Chameleon - (720-930nm)

Advantages and Techniques
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Intravital 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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