Confocal Microscopy

Instruments

The facility has two laser scanning confocal microscopes, a Zeiss LSM 510, a Zeiss LSM 510 Meta. In addition, the facility also has a combined confocal/two-photon Zeiss LSM 510NLO microscope plus two image workstations as well as other microscopy related equipment.

Zeiss LSM 510

This microscope allows the acquisition of pictures and movies from samples labeled with fluorescent probes, quantitative analysis of images and 3-4 dimensional reconstructions. This microscope has three excitation wavelengths: 488nm, 568nm and 633nm, and three detector channels.

Zeiss LSM 510 Meta

The Zeiss LSM 510 Meta allows discrimination between fluorophores with closely spaced or overlapping emission spectra due to its "meta detector". The meta feature records a complete spectrum at each pixel of an image allowing sophisticated spectral un-mixing to be performed. This microscope is equipped with 458, 477, 488, 514, 543, and 633nm excitation wavelengths and three detector channels, one of which is the meta detector. This allows the imaging of a wide variety of fluorescent probes among them green fluorescent proteins such as GFP, CFP, YFP, and Ds-Red.

Zeiss LSM 510NLO/Multiphoton Microscope

This microscope uses an argon laser and 2 HeNe lasers for excitation (at 458, 477, 488, 514, 543, and 633 nm) and also has three detection channels. This microscope is adapted to examine both fixed and live specimens. In addition, an Eppendorf pressure microinjection system is available for microinjection studies, and a fast shuttered, fiber optic-coupled UV lamp is available for flash photolysis of caged compounds.

This system is also equipped for multiphoton fluorescence imaging. Multiphoton imaging is particularly useful for imaging of fluorescence deep into tissues (up to 200 micrometers) with bleaching limited to focal planes, and for flash photolysis studies that require imaging in precise subcellular regions. The setup includes a Spectra-Physics MaiTai titanium:sapphire laser for two photon excitation. The output wavelength of the MaiTai is tunable from ~700-1020 nm. The system has also been fitted with custom-built non-descanned (external) detectors to maximize detection efficiency for imaging highly scattering specimens.

Computer workstations

Two computer workstations (Dell and Macintosh computers) are available in the facility for off-line analysis and 3D reconstruction of images. They are equipped with the LSM510 program, Volocity program for 3-4 D reconstruction and Photoshop software.

All computers are networked to facilitate data transfer for users.

Online scheduling