VISUAL SCIENCES RESEARCH CENTER

Eric Pearlman, Ph.D., Module Director

Departments of Ophthalmology and The Center for Global Health and Diseases

phone # 368-1856

eric.pearlman@case.edu

Catherine Doller, Histology Manager

Department of Ophthalmology

phone # 368-5239

catherine.doller@case.edu

Located in The VSRC Consolidated Laboratory Space in the Institute of Pathology

The Histology module provides access to technical training, technical support and equipment for researchers in the visual sciences. The Histology Module provides multidimensional support in histology that cannot be duplicated in any one laboratory. In its seven years of operation, the Histology Module has grown considerably and has become the most utilized Module in the VSRC. The Module has gone from processing 9024 slides (30498 sections) in 2001 to producing 17437 slides (53323 sections) in 2005. The module has become an indispensable component of the VSRC and is utilized widely by many laboratories on numerous projects. The histology module has become a full service research based area, often beginning with raw unprocessed tissue, which is dissected, fixed and finally sectioned to achieve the proper orientations and thicknesses needed for particular applications.

The area is fully equipped for paraffin tissue processing, embedding and sectioning, cryosectioning, and immunohistochemistry Although focusing mostly on ocular tissue, the Histology Module has the ability to process a wide variety of tissues from numerous organs and species. Routine stains such as hematoxylin and eosin and special stains including TUNEL are performed on a regular basis. Besides these stains, more complex enzyme histochemistries, and immunohistochemical techniques are also employed. The Histology Module has now become an integral part of research projects involving Laser Capture Microdisection (LCM). The Histology Module is already provides RNAse free sections for in situ hybridization, therefore it will be able to provide similar sections for LCM RNA based projects.

Equipment & Services Available

Equipment:

• -Sakura Tissue-Tek VIP Vacuum Infiltration Processor, Model E300
• -Sakura Tissue-Tek TEC Tissue Embedding Console System
• -Embedding Console, Model 4710
• -Cryo Console, Model 4709
• -Richard-Allen Microm Microtome Cryostat, Model HM500M
• -Richard-Allen Microm Rotary Microtome, Model HM355
• -Reichert-Jung Ultramicrotome, Model Ultracut E (Note: no diamond knife or glass knife cutter)
• -Shandon Automated Immunostainer, Model Cadenza
• -Triangle Biomedical Sciences, Inc. Tissue Flotation Bath, Model H-TFB
• -Precision Scientific Econotherm Oven, Model 1025-115
• -Barnstead/Thermolyne Nuova Stir Plate, Model SP18425
• -Fisher Scientific Touch Mixer, Model 231
• -Nikon Eclipse 200 Microscope

The Module Manager provides various levels of expertise depending on investigator need. The lowest level of support is to assist researchers already skilled in histological technique and involves basic training on the equipment. The highest level of support is to assist in projects from beginning to end by providing actual technical service.

Current expertise includes:

• -Gross specimen dissection and orientation.
• -Fixation techniques (frozen and paraffin).
• -Specimen processing (frozen and paraffin).
• -Specimen orientation and embedding (frozen and paraffin).
• -Specimen cutting (frozen and paraffin).
• -Routine hematoxylin and eosin staining.
• -Special stains including TUNEL
• -Mounting techniques (permanent and aqueous).
• -Production of RNAse free slides for in situ analysis
• -Production of slides for Laser Capture (LCM) based studies
• -Enzyme histochemisrty.
• -Immunohistochemistry:
• -Enzyme immunohistochemistry.
• -Direct immunofluorescence.
• -Indirect immunofluorescence.
• -Muscle biopsy techniques.
• -Tissue sectioning for laser microdissection and RNA analysis.
• -Procedure development and documentation.
• -Troubleshooting procedures.
• -Equipment maintenance.

UTILIZATION:

Use of the Module and the Module Manager's time is prioritized in the following order:

• -NEI funded projects.
• -Projects leading to NEI grant submission.
• -Vision related projects sponsored by other funding agencies.

All work in the facility requires completion of a work order form. Projects are prioritized on a first come, first served basis that is subject to adjustments in accordance with the priority sequence above. Priority is given to NEI-supported training and short-term projects with a high potential for publication and/or presentation. Lower priority is given to long-term, labor-intensive projects that are better accomplished by the training of personnel within the particular laboratory. Since this is a core facility, no one group can monopolize either the Manager's time or use of the equipment. If a project is large and requires a large amount of the Manager's time or heavy use of the equipment, a fair amount of time will be allocated to the investigator on a weekly basis so that others will also be able to schedule time. The Module Manager will have priority use on all equipment and individuals wishing to use the common equipment must schedule an appointment. The Module Director is consulted in the event of any conflicts involving the Manager's time and/or laboratory equipment.

All use of Core facility equipment requires that personnel be specifically trained and/or approved for use of the equipment by the Module Manager. All use of the Module requires an approved IACUC or IRB protocol specific to the project.

COSTS:

The facility operates on a fairly small yearly budget and provides reagents and supplies required for routine tissue processing, cutting, H&E staining and a limited number of special stains (alcohols, xylene, paraffin, microtome blades, routine stains, and routine mounting media). A limited amount of supplies such as treated slides, coverslips, OCT, etc. are provided by the lab however, if a project is large or ongoing for an extended period of time, individual investigators are required to provide these supplies. Slide boxes and/or slide trays are also required. Individual researchers will provide all immunostaining reagents other than basic buffers (primary antibodies, secondary antibodies, detection systems, special mounting media, etc.) and any specialized supplies for other procedures. This includes special slides for laser microdissection (films and foils), RNase-free reagents (DEPC water and RNase Zap or similar product) for RNA processing and microtome blades for projects with a large number of specimens (blades can only be used once). We also require individuals to provide compact disks and/or zip disks for digital photography data storage. Please refer to the VSRC Fee Schedule.