Services and Facilities
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 Diabetes Research Center 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
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.
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.
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.
Visiopharm Imaging Software with AI/Deep learning provides powerful automated tissue segmentation and image analysis. Numerous apps are available or can be developed by core staff.
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 technical assistance, training and expert consultation. Emphasis on rigor, reproducibility, validation and the proper use of controls is a key component of all core services.
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.