Cellular Imaging Core

Overview

The Cellular Imaging Core serves the CSIBD community by providing Center investigators with the technical support, expertise, and access to high-end instrumentation necessary for morphological, immuno-cytochemical, and cell biological studies related to IBD and digestive disease research.

Objectives

1. Provide access to sophisticated cellular imaging approaches that would otherwise be prohibitive to individual researchers
2. Provide an environment for training key personnel from Center laboratories
3. Evolve to meet the needs of the CSIBD community by introducing new equipment and techniques
4. Maintain the highest level of integrity in generating, analyzing, and reporting data by focusing on quality control, rigor and reproducibility parameters in research, and data management
5. Promote the development of junior investigators and foster collaborations by providing a connection point between investigators of varying research approaches

Services

• Routine tissue fixation and processing for light microscopy. The Core also performs special stains (e.g., trichrome, PAS, Sirius Red).
• Perfusion-fixation of organs. The type of fixative depends on the experiments to be carried out and sensitivity of antigens to fixation; it is determined empirically based on our experience.
• Conventional electron microscopy. Images are captured digitally using our JEOL 1011 electron microscope and AMT CCD camera. They are stored on a central file server.
• Low temperature embedding in Lowicryl HM20. Embedding tissues in hydrophilic resin preserves antigenicity better than conventional embedding and allows the post-embedding detection of antigens on thin sections of tissues, using immunogold labeling techniques.
• Immunocytochemistry. The Core performs and instructs users on a variety of immunocytochemical procedures, including the following:
• Light microscope immunocytochemistry on cells and tissue sections with 3D reconstruction
• Light microscopy of intact cells and tissues
• Electron microscopy on intact cells and tissues (pre-embedding labeling)
• Immunogold electron microscopy on thin sections
• Ultracryomicrotomy for immunogold labeling of thin, frozen sections
• Nikon A1R laser scanning confocal microscopy. The Nikon A1R confocal microscope is dedicated to live-cell imaging and associated techniques, including FRET and FRAP, and spectral imaging.
• Zeiss LSM800 Airyscan confocal imaging. This system allows image capture at close to double the resolution and much greater sensitivity compared to conventional confocal systems.
• Nikon AXR confocal platform. This microscope uses advanced hardware and AI assistance to perform all functions from image capture to final analysis more rapidly and with high sensitivity due to its high-speed resonance imaging capability.
• Airyscan imaging and Huygens deconvolution. Application of Huygens deconvolution software to Airyscan images results in greatly increased clarity and resolution.
• Total internal reflection fluorescence (TIRF) microscopy. TIRF is a powerful technique for visualizing fluorescently labeled molecules that are in or near the cell membrane.
• Calcium/pH ratio imaging. We adapted our TIRF microscope to perform Ca++ and pH ratio imaging.
• Super-resolution STORM microscopy. This system is a form of single molecule light microscopy that enables the visualization of interactions, such as between proteins or between proteins and nucleic acids, at the nanoscopic level. STORM capabilities include the use of three-color and three-dimensional imaging.
• Imaging flow cytometry. Amnis technology fully distinguishes and ­­visualizes appropriate information from undesired background debris by a combination of image analysis and brightfield examination.
• Central antibody bank. We have a bank of over 2,000 antibody reagents, many of which were developed in-house.
• Personnel training. We provide support for CSIBD investigators through (1) formal and informal end-to-end consultations, (2) short-term technical training, and (3) intensive training of selected personnel in specific activities that will be used extensively by participating laboratories.