The mission of this core is to facilitate analysis of the regulatory factors in the GI tract both in vivo and in vitro, using a variety of microscopic techniques, and to correlate structure with function by using light, confocal and electron microscopy. A new offering is laser capture microdissection, for characterizing mRNA and protein expression and profiling intermediary metabolism in specific cell populations in the mouse intestine. This core performs a large array of services for its users, including routine paraffin and frozen section for gastrointestinal and other tissues, histologic stains, a wide variety of immunohistochemical techniques, confocal microscopic, electron microscopic, and laser capture microdissection services. This core has optimized immunohistochemical and immunocytochemical staining techniques for a large number of antibodies, using avidin-biotin, routine and double immunofluorescence and tyramide enhancement methods. Functional studies offered by the core include TUNEL assays for apoptotic cells and immunohistochemical detection of 5'bromodeoxyuridine in proliferating cells. Dr. Rubin provides data interpretation services for individual investigators, and training for all components of the core is available for our users.

SERVICES OFFERED:

Investigators are encouraged to contact Deborah Rubin for information about services listed, for potential new services or for other consultative information.

HISTOLOGY COMPONENT

The Histology component is located in the CSRB North Tower Addtion Room 924. This component handles and processes all types of tissues, including making paraffin or OCT (frozen) tissue blocks, sectioning, a wide variety of special stains, and single, double and triple immunohistochemical procedures. Users of the Histology component also have access to two imaging systems, a Zeiss Axioskop 2 MOT microscope with full light and immunofluroescent capabilities (4 color fluorescence), polarizing filter for crystal analysis, attached to an Axiocam MRM Digital camera that takes black and white images that can be analyzed for multilabel immunofluorescence, and a Zeiss Axioskop 40 microscope attached to an Axiocam MRC digital colar camera, both with Pentium III and IV computers and Axiovision image analysis software. We have developed detailed protocols for optimization of immunohistochemical procedures for antibodies provided by the investigators.

CONFOCAL LASER IMAGING COMPONENT

The confocal laser imaging component contains a Zeiss Upright microscope with BioRad MRC 1024 confocal adaptor capable of 3 color fluorescence, and a new Zeiss Confocal Microscope LSM510 with a Meta adaptor. This component is located in the Dept. of Cell Biology. Dr. Robert Wilkinson is the director of this component. A full time technical expert is available to train and assist investigators in the use of the confocal microscope and workstation.

ELECTRON MICROSCOPY COMPONENT

The electron microscopy component offers specimen preparation including fixatives, thin sectioning, negative staining, grid preparation, metal evaporation, electron microscopy and photography. It is located in the Dept. of Cell Biology, and is administered by Dr. Philip Stahl. Investigators may train with Marilyn Levy, senior electron microscopy technician, in the use of the electron microscope and photography, Equipment includes a Hitachi H-600 transmission electron microscope, a Hitachi 5-450 scanning electron microscope, an ultramicrotome and other standard equipment for tissue preparation and sectioning.

LASER CAPTURE MICRODISSECTION COMPONENT

The laser capture microdissection component will train investigators on using the Arcturus PixCell II system, located in the laboratory of Dr. Jeffrey Gordon. Mr. Bill Coleman is charged with training users.

Please Note:

Because the demand on services exceeds our core's capacity, no new work will be accepted from non-DDRCC members. Job assignments will be prioritized by the membership criteria (full, associate and affiliate).