SPECIALIZED DEPARTMENTAL RESEARCH FACILITIES
Confocal Microscope
The department has recently acquired a state-of-the-art Laser Scanning Confocal Microscope Imaging System obtained through a Shared Instrumentation Grant from the National Institutes of Health. The instrument is located in Room 814 of the Molecular Sciences Building and is a Zeiss LSM 510 microscope configured on an inverted Zeiss Axiovert 100M base. This configuration allows investigators to examine specimens in tissue culture dishes for live cell, real-time fluorescence studies, as well as samples prepared on conventional glass slides. Investigators who require that their samples be maintained at physiological temperatures can be accommodated since the microscope also has a temperature regulated stage which allows the user to acquire images from live cells maintained at temperatures between 10oC and 45oC.
The optics and laser systems were chosen to accommodate a wide range of user preferences. The system is equipped with three (3) lasers with which wavelength selection and attenuation are handled by Acousto Optical Tunable Filters (AOTFs). The argon ion laser (458 nm, 488 nm, and 514 nm), 1 milliwatt helium-neon (HeNe) laser (543 nm) laser, and 5 milliwatt 633 nm HeNe laser allows users to chose from a variety of fluorochromes that require excitation from 458 nm to 633 nm, including the most commonly used dyes and fluors such as lucifer yellow, wild-type and enhanced GFPs (green fluorescent proteins), fluorscein, rhodamine, Cy2, Cy3, Cy5, Cy7, Texas red, and phycoerythyrin. The instrument has three PMT’s, each with it’s own pinhole, for simultaneous three-color fluorescence image acquisition. Samples can also be examined, and images acquired, using DIC (direct interference contrast) optics in the nonconfocal, transmitted light mode. The objectives were chosen also to provide a wide range of magnifications to accommodate a variety of applications. These include a Fluar 5X/0.25, a Plan-Neo 10X/0.3, an Achroplan 40X/0.6 -long working distance, a Plan-Neo 40X/1.3, and a Plan-Apo 100X/1.4-Oil.
The entire system is controlled by user-friendly, menu-driven software which is run on a WINDOWS NT 4.0-operating system driven by a 400 MHz Pentium II processor. The two 21-inch monitors provide maximum flexibility for viewing images or for simultaneous image acquisition and data analysis. The software package includes programs for both 2D and 3D image processing, as well as more sophisticated operations such as acquiring Z-stacks from multiple cells over time, uncaging experiments, and photobleaching studies. Images can be collected at resolutions up to 2K x 2K pixels and with up to 12 bit digitization. An additional workstation is also available for post-acquisition data processing or image reconstruction and analysis.
A full-time technician is available to help first-time users obtain images quickly with minimal down-time for training, as well as to provide technical assistance for more experienced users.
Reservations:
Reservations can be made up to a week in advance by accessing the Confocal Reservations Web-Site, or by signing up on the reservation sheet on the door to Room 814 in the Molecular Sciences Building, 858 Madison Ave.
Phospho-imager
The department is equipped with a Molecular Dynamics Storm 860 System for gel and blot analysis that supports radioactive PhosphorImager system technology, as well as two non-radioactive fluorescent labeling techniques. Storm uses storage phosphor screens instead of film to capture quantitative data from radioactive gels and blots. The linear dynamic range of this system is 1000 times greater than film allowing direct comparison and quantitation of signals that varying greatly in intensity. Screens available in the department allow the use of many commonly used isotopes such as 32P, 33P and 35S. This system also supports non-radioactive techniques. You can detect nucleic acids or proteins on gels using direct fluorescence, as well as detect nonradioactive DNA, RNA and proteins on blots using chemifluoescence techniques.
The raw data are analyzed by ImageQuant software. This software uses push-button tools to select software objects and functions for defining and analyzing gel and blot data. In addition we have a variety of other software programs including Fragment Analysis that performs automatic DNA fragment sizing, calculates protein molecular weights and analyzes isoelectric focusing gels.
This instrument is located in Room 821 of the Molecular Sciences Building. Storm System allows the analysis of data from many diverse applications including: Northern blotting, Southern blotting, Western blotting, band shift assays, CAT assays, kinase assays, library screening, 1-D and 2-D protein gels, mini-array hybridization and many others.
Epifluorescence Microscope
The department
has a Zeiss Axiophot microscope equipped with epifluorescent optics. This
microscope is located in Room 601 of the Molecular Sciences Building. This
microscope allows investigators to examine live cells in tissue culture dishes
for real-time fluorescence studies, as well as fixed samples prepared on
microscope slides. The microscope
has a number of objectives including a 63x PlanApochromat, a 40x Plan-Neofluar,
a 25x Plan-Neofluar, and a 10x immersible lens. Images obtained with the
microscope can be captured with an Optronics digital camera that is connected to
a Windows 95 driven computer work station.
Computers
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