Image Acquisition & Analysis Module

Director: Ian A. Sigal, PhD

Purpose

The Electronics/Molecular Imaging module is dedicated to providing hardware and software support for acquisition, analysis and storage of digital microscopic images.

Laser Scanning Confocal Microscopy

Laser scanning confocal microscopy addresses several of the problems encountered in traditional microscopy by controlling the depth of filed, eliminating image degradation through out-of-focus information and by allowing the collection of serial optical sections from thick specimens.

Our confocal system, an Olympus Fluoview1000 is equipped with an Olympus IX81 inverted microscope, offers mercury lamp excitation for initial viewing and four lasers with six possible wavelengths of laser excitation for confocal acquisition. It has 10x, 20x and 60x air objectives with phase contrast available as well as 20x, 40x and 60x and 100x oil objectives.

Live Cell Microscopy

Live cell microscopy allows researchers to observe and quantify phenomena in living cells via technological advancements in hardware, software and laboratory techniques. This approach to microscopy requires creation of an environment hospitable to maintenance of cell life for the duration of the experiment.

Our live cell system revolves around the Nikon Eclipse TE200-E inverted microscope with 4x, 10x and 20x air objectives and 40x and 60x (TIRF) objectives. Illumination is provided by a DG-4 xenon light source and a wide variety of excitation and emission filters are available. Peripherals include an Eppendorf microinjection system, a Roper CoolSnapHQ camera, uniblitz brightfield shutter, a heated objective, stage and chamber cover, x, y and z automated stage control and a suspension table. Acquisition and analysis are performed in MetaMorph.

Services

  1. Provide training for the operation of a digital imaging microscope for stationary specimens; a confocal microscopy system; and a microscope system for automated real-time imaging and micromanipulation.
  2. Provide assistance with the techniques of acquiring and analyzing digital light, fluorescent, and confocal microscopic images, and time lapse, z-series and multiple stage position images using the MetaMorph automated imaging system.
  3. Serve as a resource for information on multiple analytical applications of different imaging software currently available at the core imaging facility, and advising on upgrades for future needs and applications.
  4. Diagnose and trouble-shooting problems with the imaging microscopes, micromanipulator and gel analysis systems.
  5. Integrate data from various imaging systems by designing or implementing software allowing importation of data into MetaMorph for more sophisticated analysis than that available in the software packages that accompany the instrument.

Citation

Please cite the Core Grant for Vision Research - EY08098 in your publications if you have used the services of this module.

Contact Information

Kira Lathrop
412-647-3492
kll21@pitt.edu
EEINS-1026, 203 Lothrop Street
Pittsburgh, PA 15213