Flow Cytometry Module

Director: Robert L. Hendricks, PhD

The Morphology Module provides technical support for researchers requiring sample preparation for light, confocal, and electron microscopy. These services include fixation of tissues and cultured cells, embedding, section preparation, and staining. The module will also provide facilities and training for individual researchers who prepare and stain their own samples and the module maintains a library of commonly used antibodies and histological reagents available to all core grant users.

Equipment

The lab contains a Leica 2800 Frigocut-E cryostat, Microm HM505E cryostat, a fume hood, light microscope, dissecting scope, refrigerator-freezer and histological technique references. The 10th floor laboratory maintains a library of secondary antibodies labeled with fluorescent tags compatible with filter sets available in the Imaging and Image analysis Module. The Histology Core (2nd floor) provides an Olympus 4060E microscope, a Lancer Vibratome Series 1000, Reichert Jung Autocut Microtome (paraffin & plastics), a Leica-Jung Polycut E fully automated microtome for large sections, a new Leica CM30505 cryostat, a Leica 2145 Microtome, Leica SM 2000R semi automatic sliding microtome, 2 AO Spencer 860 sliding microtomes and 4 AO Spencer 820 rotary microtomes. For electron microscopy there is a JB-4, and 2 LKB ultramicrotomes and an LKB glass knife breaker. For tissue processing a Tissue Tek VIP processor and a Tissue Tek embedding processor are available. A full range of standard histological stains are available. The Core Facility also has access to major equipment such as –80º C freezers, a microwave processor for microwave fixation and antigen recovery methods and a thermal cycler for in situ polymerase chain reaction methods, located on the first floor of the Eye & Ear Institute.

Services

The module technician maintains equipment and reagents in the module laboratory. She works directly with individual researchers advising them on the appropriate fluors for the available microscope filters and secondary antibodies. She carries out fixation, sectioning, and staining of submitted samples. Staining can include standard histological stains, immunofluorescent, or in situ hybridization. The technician will also teach histological procedures to researchers that want to carry out procedures themselves. The technician is responsible for keeping records of services performed and meets on a daily basis with the module director. The module technician coordinates projects with the technicians of the Hybridoma and Tissue culture modules as well as the Molecular Biology Module for use and testing of reagents (such as monoclonal antibodies and labeled probes) common among the modules.

Contact Information

Nancy Zurowski
412-647-1413
nbz1@pitt.edu
EEINS-1048, 203 Lothrop Street
Pittsburgh, PA 15213