About the Core
Tissue morphology and knowledge of the localization of specific proteins within a cell are integral to understanding cellular metabolism and the causes and consequences of diabetes and new advances for the treatment of diabetes. The tools of morphological analysis are varied but most involve particular skills or equipment that are too costly to be supported in individual laboratories. The Advanced Microscopy Core provides a wide range of service covering the broad categories: performing specific morphological procedures, providing access to equipment and reagents, and giving advice, training and interpretation. The effort in all the facilities is to provide prompt, customized service.
There are five main components in this core: Electron Microscopy (EM), Confocal Microscopy, Histology and Laser Capture Microdissection (LCM) facilities and Morphological Consultation. The components run with different operation schemes: the plastic embedding and electron microscope work is done by the Core Manager; he also does the highly specialized immunolocalization of specific molecules at the ultrastructural level. The Core Histology Technicians take fixed/frozen tissue prepared by users and return slides (H&E and unstained for immunohistochemistry). Both the Confocal microscopes and Laser Capture Microdissection (LCM) are used directly by the individual investigators with the Core training new users, providing maintenance of the microscopes, and troubleshooting problems. Mr. Cahill trains the users for both the Confocal and LCM equipment.
Services of the Advanced Microscopy Core:
1. Tissue preparation / embedding / sectioning
a. Paraffin embedding and sectioning: Tissue is fixed in each investigator’s lab; the Core serves as a resource of information about fixatives and procedures. An order is initiated online.
b. Cryostat sectioning/use of cryostat Frozen blocks of tissue are prepared by the investigator and brought to the core freezer; specific instructions can be included on the on-line order form. After training, researcher can book the cryostat for individual use.
c. Plastic embedding and sectioning for Light microscope A useful service for very small samples, such as cell pellets, transplanted islets, pituitary grafts or young mouse embryos, for light microscopic analysis. Order is made under EMNet online.
2.Transmission electron microscope analysis Samples analyzed on EM are first fixed, osmicated, embedded and sectioned as above for plastic sections. Fixation procedures are discussed with the Core personnel; fixatives and their recipes are available from the core. Immunochemistry on ultrastructural level is also available. Order is made under EMNet online
3. Confocal Microscopy Use After training, researchers can use independently with sign up on Intranet.
a. Zeiss LSM 410 for routine imaging of immunofluorescent with 3 channel (red, green, and blue UV).
b. Zeiss LSM710NLO This microscope with 6 laser lines including two photon, 3 spectral detection channels allows multifluorescent spectral imaging, ion imaging, enhanced 3-D examinations, and manipulations for FRET or FRAP. Live cell imaging is possible.
4. Laser capture microdissection facility use. Arcturus Pixcell II Laser capture microscope and accessories. After being trained, researchers use it independently.
5. Consultation Expert advice provided by the Core personnel
Chris Cahill, Core Manager
Alevtina Pinkhasov, Core Technician
Anna Monisva, Core Technician