The Confocal Microscopy Facility (CMF) is located in RRC-West of the University of Illinois at Chicago.

Zeiss LSM 510 and LSM 510 META confocal microscopes were installed in 1998 and 2003 respectively. They 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. 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. Both microscopes also have a transmitted DIC channel.

There are differences, however, between the LSM 510 and the LSM 510 META:

1. The available lasers for the LSM 510 are: Ar/Kr 488/568nm laser for green and red dyes; Ar/UV 351/364nm laser for blue dye; and HeNe 633nm laser for dark red dye.

   The LSM 510 META is equipped with: Ar 458/488/514nm laser for cyanine, green and yellow dyes; HeNe 543nm laser for red dye; HeNe 633nm laser for dark red dye and Diode 405nm laser for blue dye.

2. 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.

3. 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.

An AIS2 automatic microinjection system was installed in November 2001. The investigator can operate the system entirely via the computer workstation and inject cells 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 between 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.