Confocal Microscopy Facility (CMF)
The Confocal Microscopy Facility at RRC UIC provides services for the study of biological processes at the tissue and cellular levels. The services include light, deconvolution, confocal and TIRF microscopy, an AIS2 microinjection system, temperature controlled stage for live cell observation, and workstations for digital image manipulation and analysis.
Overview
The Zeiss LSM 510 and LSM 510 META confocal microscopes are designed for analysis of living or fixed biological specimens in a multi-user environment. Using the "Z-Stack" software module, both microscopes can scan through a sample and obtain one or more high resolution optical sections. Users then have the option of utilizing a Z-Stack to render a 3D reconstruction of the sample. The microscopes can provide images of live or fixed cells and tissue sections of varying thickness. The time-lapse imaging function can be used for kinetic studies of live cells with GFP, calcium indicator, FRAP applications, etc. Since both microscopes use the same software packages, there is no retraining necessary.
The LSM 510 is equipped with four separated confocal channels for detecting the blue, green, red and dark red dyes simultaneously without any bleed through. An advanced function of the Zeiss LSM 510 META microscope is the multi-tracking scanning capability that minimizes signal crosstalk while working with multiple fluorescences. The META detection module of the LSM 510 META provides fast acquisition of image stacks with spectral information for every pixel. With its emission fingerprinting technique, it permits the clean separation of several, even spectrally overlapping, fluorescence signals of a specimen such as the separation of GFP and YFP signals. The laser and the configuration settings are suitable for FRET. Both microscopes also have a transmitted DIC channel.
An AIS2 automatic microinjection system is operated entirely via the computer workstation. Cells are injected by pointing and clicking which results in high injection rates of about 1,500 cells per hour. The injected cells can be marked for relocating and avoiding double injection. A unique aspect of our AIS2 system is its combination with the LSM 510 META confocal microscope, which allows the user to trace interesting targets with the time-lapse program. These two systems can be rapidly switched by clicking one button. The combination of confocal microscopy and microinjection techniques bridges the gap between in vivo physiology and in vitro biochemistry and molecular biology. Investigators can study complex cellular processes, structure and function in vivo using single cells.
Contact the director of the facility, Dr.Mei Ling Chen to make an appointment to discuss experimental procedures or sample preparation. New users: Complete an application form signed by P.I. and schedule training by contacting Mei Ling. An on-line reservation system is under development