FACILITIES: UW HISTOLOGY AND IMAGING CORE (HIC) AT SOUTH LAKE UNION
Laboratory: The Histology and Imaging Core (HIC) encompasses approximately 2,800 sq ft on the 3rd floor of building B at the South Lake Union (SLU) campus of the University of Washington (UW) School of Medicine. The HIC is a UW resource that is run under the auspices of the Department of Comparative Medicine (DCM). The HIC has 2 offices, bench space and desks for 10+ research scientists and histotechnologists, imaging suites with state-of-the-art microscopes, access to animal procedure rooms, image analysis workstations, and a small conference room for comparative pathology consultations.
Animal: Animals are housed in two modern specific pathogen-free, barrier facilities at the SLU campus. An excellent staff including clinical veterinarians and veterinary pathologists, manage this facility through the Department of Comparative Medicine.
Computer: The HIC has a variety of late-model personal computers, both Macs and PCs, image analysis workstations, scanners, and printers. Data and files are backed up over a locally networked drive daily. In addition, the HIC has a network (ImagePath) that includes a 32 terabyte file server for storage of image files and data generated in this resource.
Office: Core Director, Charles W. Frevert, DVM, ScD and Co-Director, Piper Treuting, DVM, MS, DACVP, are comparative pathologists with faculty appointments in the DCM. Both Drs. Treuting and Frevert have offices in the HIC. The Department of Comparative Medicine provides administrative support for grant management, personnel management, website, and oversight of the cost center.
Other: Drs. Frevert and Treuting have independent research projects in the Comparative Pathology Program, which is centered at the University of Washington at South Lake Union. They also oversee the University of Washington Veterinary Diagnostic Laboratory and the veterinary pathology space at the UW for the Department of Comparative Medicine. Brian Johnson (email@example.com), a senior research scientist, with extensive experience in histology, immunohistochemistry and imaging is the Program Manager for the HIC.
|MAJOR EQUIPMENT AND RESOURCES AVAILABLE THROUGH THE HIC|
HIC Overview: The HIC is a UW resource for the analyses of biological samples, cells and tissues, which is open to all investigators, including; UW and their affiliates, commercial and not-for-profit biotechnology companies, and other academic institutions. As a pathology laboratory for experimental and preclinical studies, the HIC offers state-of-the-art services related to histochemical analysis, immunohistochemistry, microscopy, image acquisition and quantitative microscopy as well as routine services such as tissue processing and embedding (Table 1). Whereas several of the services are routine, personnel in the HIC work with investigators to provide strict and necessary quality control at all steps of the process. Other critical services offered by the HIC include expert technical assistance, consultation, comparative pathology and training.
Comparative Pathology: A unique strength of the HIC is access to veterinarians who have expertise in comparative pathology. Charles Frevert, the Director of the HIC and a veterinarian trained in comparative pathology, works closely with three board-certified veterinary pathologists at the UW: Piper Treuting, Jessica Snyder and Denny Liggitt. Dr. Treuting is recognized nationally and internationally as a mouse pathologist. Dr. Snyder is a specialist in comparative pathology and veterinary neurology with a focus on neuropathology. Dr. Liggitt, Chair of the Department of Comparative Medicine, has broad experience in biomedical research working in both the academic and private sectors. When necessary, consultation with Drs. Treuting, Snyder, Liggitt and Frevert, will be available to investigators working with animal models of human disease. Having access to board-certified veterinary pathologists through the HIC also provides the critical expertise required for assisting with preclinical studies, especially preclinical toxicology analysis for investigational new drug candidates.
Histology: The HIC has standard lab equipment (freezers, refrigerators, microfuge, scales, shakers, ovens, etc.), and histology equipment. To increase productivity in the core, much of the histology equipment is automated. This includes Brady slide and cassette labelers, a Tissue-Tek VIP automated tissue processor, Leica embedding station, various microtomes, a Leica motorized microtome for paraffin and plastic, a Leica cryostat, a Leica automated stainer and a Leica automated glass-coverslipper. The HIC provides H&E stains as the standard for all cases processed in the core. Upon request the HIC can perform a number of affinity based histochemical stains including Trichrome, Sirrius Red, Giemsa, PAS, PTAH, Movat’s pentachrome, tissue gram-stains and silver stains.
Automated Immunohistochemistry: The Leica Bond is an automated system for immunohistochemistry (IHC). This system includes deparaffinization, cell conditioning, and staining and allows investigators a great deal of flexibility employing varied incubation times and temperature settings to optimize their protocols. This instrument is used for the development of new IHC protocols as well as running standardized protocols that have been developed by HIC staff. Information on the antibodies that have been optimized for immunohistochemistry on human and mouse tissues can be obtained at the HIC website or by contacting Brian Johnson, the Program Manager (firstname.lastname@example.org).
Hamamatsu NanoZoomer Digital Pathology (NDP) System: This unit is an automated high-resolution slide scanner that scans and digitizes tissue sections on glass slides in batches of up to 210. The digital images can be visualized over the internet and used for morphometric analysis. Automated image analysis can be performed on digitized slides using Visopharm® image analysis software
Image Analysis: For the majority of the quantitative microscopy performed in the HIC, protocols are developed using Visopharm software and whole digital slides generated with the Hamamatsu NDP imaging system. In addition, the HIC has 2 Dell image analysis workstations running a number of imaging programs including Visopharm Image Analysis Software, Image ProPlus Software, Nikon Image Analysis Software (NIS Elements), NIH ImageJ Software and Adobe Photoshop. These platforms will allow investigators to perform image analysis away from the microscopes (e.g., Hamamatsu NanoZoomer, Nikon 90i or DeltaVision) to capture digital images.
Epifluorescence and Bright Field Microscopy: This motorized Nikon Eclipse 90i has high-end CFI plan apo-objectives, differential interference contrast (DIC) capabilities for the 20, 40, 60, and 100x objectives for bright field and fluorescence microscopy. In addition, there are two digital cameras – a color (Nikon 12 mega pixel) and monochromatic (Coolsnap ES2) digital camera – plus NIS Elements acquisition and analysis software.
Live-cell imaging: For live cell imaging, investigators have access to a DeltaVision Elite inverted wide-field deconvolution microscope. This microscope has dual laser-illumination and proprietary motion-control technology as well as deconvolution software. The DeltaVision microscope is particularly useful for gathering low-intensity signals, which sets it apart from the confocal microscope. This system also provides investigators the ability to perform live cell imaging in four dimensions, x, y, z, and time. In addition, when performing imaging for thin sections (5 microns to 20 microns) of tissues or cellular preps prepared for fluorescent immunohistochemistry or immunocytochemistry the DeltaVision Core offers several advantages over the confocal microscope including the ability to capture low-intensity fluorescent signals as well as significantly better resolution in the Z axis.
Multiplex Bioassays: A Luminex Bio-Plex 200 suspension array system with high-throughput fluidics and an automated microplate wash station is available in the HIC. The reader combines two lasers, fluidics, and real-time digital signal processing to distinguish up to 100 different sets of color-coded magnetic or polystyrene beads, each bearing a different antibody using a standard 96-well microplate format. Microplate processing and data acquisition and analysis services are available from HIC staff.
ImagePath Network: The ImagePath Network, which is located and administered through the HIC, is comprised of a domain and file server. The file server has 32 terabytes of file storage for digital images and data generated in the HIC. In addition, the VADDs database, which is used for monitoring workflow and billing within the HIC, is located on the ImagePath Network.
|Table 1: HIC Services and Technologies – Histology and Imaging Section|
|Spatial Localization of Proteins
||Spatial Localization of mRNA
|Live Cell Imaging
||Image Analysis and Morphometry
|Microscopy and Digital Imaging
Extended Training Workshops and Short Courses Offered Through the HIC: Piper Treuting, Charles Frevert and Brian Johnson are course faculty and help to direct three internationally recognized courses that are offered as extended training in mouse pathology, immunohistochemistry, imaging and quantitative microscopy.
Annual Workshop on the Pathology of Mouse Models for Human Disease: This annual workshop provides a week of intensive training sessions in pathology and histopathology as well as didactic sessions in which particular disease areas and models will be discussed. Participants will have an opportunity to interact with a group of prominent mouse pathologists and geneticists from leading research institutions. Additional information can be found at the workshop website.
The Immunohistochemistry and Microscopy Course (IHCM): The IHCM course, held annually in March, is four full days and evenings (11 hours daily) of lecture and laboratory sessions with experts in the field of immunohistochemistry (IHC) and microscopy. The IHCM course goal is to provide participants in-depth theory of and extensive hands-on experience with a broad range of immunohistochemistry (IHC) and microscopic imaging techniques. The course emphasizes hands-on laboratory time and small breakout discussions with faculty and staff. Additional information can be found at the special topics course website.
Workshop on Quantitative Microscopy: This workshop, which is given every two years at the University of Washington-South Lake Union, is an intensive study of the techniques and principles used in stereology and image analysis. This workshop provides a unique opportunity to interact with experts in the field of stereology and image analysis. In addition, students will be introduced to processes that will dramatically improve efficiency and accuracy of stereology and image analysis studies. The workshop is five days of lectures, interactive sessions and hands-on experience. Additional information can be found at the workshop website.
Personnel: All personnel in the HIC are full time employees in the Department of Comparative Medicine.
Charles Frevert, DVM, ScD
Director of Histology and Imaging Core
Director of Comparative Pathology Program
Piper Treuting, DVM, MS, DACVP
Director of Comparative Pathology Program
Co-Director of Histology and Imaging Core
Brian Johnson, ASCP-PA
Program Manager/Senior Research Scientist: Brian has over 10 years experience in immunohistochemistry, imaging and quantitative microscopy. He has been certified as a pathologists assistant (ASCP-PA) and has considerable experience with all aspects of molecular pathology.
UWVDL Program Operations Manager: Kerrie is a fiscal specialist who oversees the University of Washington Veterinary Diagnostic Laboratory (UWVDL). She works closely with Brian Johnson in overseeing the HIC budget and is responsible for managing VADDs, the medical management software used for accessioning of HIC and VDL work orders and providing clients with reports and invoicing services.
Tim Birkland, PhD
Research Scientist/Imaging Specialist: Dr. Birkland has over 20 years’ experience with in vitro and in vivo studies. He provides assistance with preparation and imaging of cells and tissues in the HIC.
Research Scientist/Cell biology: Tara Wigmosta is responsible for conducting ELISA, quantitative RT-PCR and multiplex assays for cytokine/chemokines and other proteins required by the research base. She will also assist with chemotaxis assays, cell isolation, immunohistochemistry and live-cell imaging.
Research Scientist/Imaging Specialist: Megan is the Visopharm specialist in the HIC and assists the research base with quantitative analysis of tissues stained with special stains or immunohistochemistry. She also assists with imaging on the Deltavision Elite and Hamamatsu Digital pathology systems.