Washington Cellular and Molecular Imaging Core

Overview

The Cellular and Molecular Imaging Core facilitates and enhances the research of Diabetes Research Center affiliates by providing access to specialized expertise and equipment required for high quality histochemical and morphological analyses in a cost-effective manner.

The Cellular and Molecular Imaging Core provides Diabetes Research Center affiliate investigators with:

• Rigorous performance and quantitative assessment of immunohistochemistry and morphological changes in tissues;
• Access to state-of-the-art imaging instrumentation;
• Expert diabetes-related technical and scientific support, including design and interpretation of research protocols;
• Access to comparative pathologists, to assist with experimental and/or preclinical studies; and
• Hands-on and didactic training Hands-on and didactic training for routine and highly specialized morphological, histochemical, and imaging techniques.

Services

The Cellular and Molecular Imaging Core has two main locations: the Cellular and Molecular Imaging Core at VA Puget Sound Health Care System and in the Histology and Imaging Core at UW SLU. These two laboratories provide state-of-the art facilities to DRC Affiliates and enable the following services to be offered:

Conventional Bright Field and Fluorescence Microscopy

Nikon NiE (VA site) upright motorized microscope configured for bright field (including DIC) and 4-color fluorescence (LED; AURA II), large field of view monochrome and color cameras, and high NA dry and oil immersion objective lenses.

Nikon Ti2 (VA site) inverted microscope with 5-color LED (AURA III), large field of view monochrome and color cameras, and high NA dry and oil immersion objective lenses.

Nikon 90i (SLU and VA sites) microscope configured for bright field, DIC and multichannel fluorescence.

Digital Pathology

Hamamatsu NanoZoomer Digital Pathology Virtual Microscope (SLU site) scans whole slides and digitizes tissue sections. Images can be viewed over a network or the internet and imported into Visiopharm software for image analysis.

Confocal Microscopy

Nikon A1R HD dual scanning confocal microscope (VA site), is equipped with a high speed resonance scanner (15 fps for 1024×1024 full frame scanning or 420fps in band scan mode) and high resolution Galvano scanner. The system comprises 4 laser lines (405nm, 488nm, 561nm, 640nm) and numerous high numerical aperture dry, oil and water immersion objective lenses.

Super-Resolution Microscopy

Nikon N-SIM E (VA site)wide-field system offers lateral resolution of 115 nm and an axial resolution of 269 nm. This system includes 3 laser lines (405nm, 488nm, 561nm) and is compatible with standard labels and dyes.

Nikon N-STORM (VA site) provides 20nm lateral resolution and 50nm axial resolution for imaging using synthetic photo-switchable dyes with a specific buffering protocol. Specific features include high speed acquisition (up to 500Hz), 4 laser lines (405nm, 488nm, 561nm, 640nm) and utilizes the best available 100X TIRF objective lens.

Laser Capture Microdissection

Leica LMD7 includes high powered dissection laser, linear-encoded scanning stage (100 nm XY repeatability) and collection from up to 3 slides simultaneously. System is equipped for bright field and DAPI, GFP, CFP, TRITC and Cy5 fluorescence. A range of objectives (1.25 – 63X) and collection devices (including 96-well plates) provides substantial flexibility.

Live Cell Imaging

DeltaVision Elite (SLU site) and Nikon Ti2/High Content (VA site) systems are equipped for live-cell imaging as well as conventional imaging for multichannel fluorescence.

Image Analysis

Nikon NIS Elements integrates image acquisition, visualization and analysis in one platform. Visualization and analysis of conventional, confocal (z-stack) and time lapse datasets including deconvolution are available. Several automated image analysis routines are available and can be customized for new projects.

Tissue Processing and Embedding

• Training and consultation in proper tissue fixation, processing and embedding
• Tissue processing (Tissue-Tek VIP and Leica ASP300S)
• Paraffin embedding (Leica EG1150H and Histocore Arcadia H/C)
• Paraffin sectioning (Leica RM 2035, RM 2065 and RM2255)
• Cryostat sectioning (Leica CM1950 and CM3050S)

Histochemistry and Immunohistochemistry

• Leica BOND RX and BONDMax for automated immunohistochemistry (chromagen, fluorescence and multiplexing). Includes on-slide antigen retrieval for superior reproducbility
• Leica Autostainer XL for H&E and other histochemical stains

Technical Assistance and Consultation

Core directors and staff provide consulting services to affiliate investigators that use the core facilities on protocols needed to meet their scientific objectives, as well on procedures for analyzing and interpreting the results of these methods.

Comparative Pathology

Access to veterinarians with expertise in comparative pathology, to assist Diabetes Research Center affiliates in translating their findings in animal models to human disease. Contact Charles Frevert for more information.