COM4S_M.TTF

The Microscopy Core provides researchers with the ability to perform both live and fixed cell multidimensional microscopic analysis and microdissection with state-of-the-art instrumentation. The Core offers expert technical assistance in microscopy and image analysis, training in advanced microscopy, and the development of novel protocols for imaging and live cell video analysis. The Core operates the following shared instruments:

Leica AS MDW.

A dedicated workstation for multi-dimensional live cell imaging incorporating a piezo z-positioner, Xenon light source, and monochromator. The software includes a workflow oriented acquisition interface, as well as deconvolution and 3-D rendering software. The MDW system has a high precision, motorized stage to permit multi-position time-lapse imaging. The 63x live cell objective is optically optimized to function at 37ºC. Other cobjectives of the Plan-Fluotar and Plan-Apo classes are incorporated in the systems offering the ability to perform Nomarski DIC and phase contrast in the range of 10x to 100x. The system incorporates a speed board to synchronize all functions to reduce acquisition idle times to the absolute limit; this permits even fast cell dynamics to be recorded in 6-D. The instrument is equipped with environmental chamber for long-term cell maintenance at 37ºC and desired CO2 concentration and has automated DIC and fluorescence capabilities with an extended range monochrome camera and Leica FW4000 software for state-of-the-art 3-D imaging.

Zeiss LSM 510 META Confoal Microscope.

Confocal microscopy is distinct in that the system limits data collected to the light emitting or reflecting from a single plane of the sample. This technique is useful in taking Z-Series images, where multiple scans are taken in increments along the Z-axis of the sample. Includes Zeiss Axiovert 200 inverted automated microscope with fluorescence, HeNe 1 & 2 and Argon/2 lasers, antivibration table, the LSM 510 META Detector, and LSM 510 software. Zeiss Axiovert 200 comes equipped with 10x, 25x, and 63x objectives. Images can be viewed in bright field, phase, DIC, or fluorescence from wavelengths of 543 nm, 633 nm, 458 nm, 477 nm, 488 nm, and 514 nm. Using the concept of Emission Fingerprinting, the LSM 510 META detector is able to separate even widely overlapping emission spectra, allowing for more precision in fluorescent imaging. Fluorochromes that resemble other emission spectra peaks can be used simultaneously through the technique of linear unmixing. Time lapse acquisition is another widely used feature on the confocal microscope, and can be combined with the Z-series function for greater accuracy in data analysis.

Zeiss PALM® MicroBeam System with non-contact laser microdissection and pressure catapulting

This system for non-contact micromanipulation and microdissection of tissue sections and live cells is using a high-photon density nitrogen UV laser, which can precisely cut out a selected area, and catapult isolated cells directly into the cap of a small tube for subsequent evaluation by PCR or any other methods. The PALM® Micro Tweezers use red or near infrared lasers to catch and move small objects microscopically with ease and accuracy. The microscope is fully automated with 10x BF/Phase, 40xLD BF, 63x BF (oil), and 100x BF (oil) objectives. Fluorescence is detectable for green (excitation 475nm-495nm BP, emission 515-565nm BP) and red (excitation 510-560nm BP, emission 590nm LP). Images are transferred to the monitor in “real time,” thus allowing for manipulation of all computer components through the software interface. The Palm Robo software can map out regions for collection, or the user can utilize the mouse and footpad accessories to perform the microdissection manually.

Instrument	Model	Applications	Contact Person
Light microscope	Leica AS MDW	Multi-channel 4-D live cell imaging and video microscopy, deconvolution	Christine Hanko
Confocal microscope	Zeiss LSM 510 META Confocal Microscope	Multi-channel 4-D imaging of cells and tissue sections, deconvolution
Microdissection	Zeiss PALM® MicroBeam System	High precision microdissection and microcapturing