The purpose of the Morphology and Metabolic Analysis Core is to improve the efficiency of diabetes-related research through provision of services that optimize the investigational capacity of Diabetes Research Center (DRC) members. Services are provided for tissue pathology and cellular architecture and function in order to assist members in investigations of the etiology and consequences of diabetes.
Fumihiko Urano, M.D., Ph.D.
Samuel E. Schechter Professor of Medicine
Office Location: 8808A Wohl Hospital
Mailing Address: Campus Box 8127
660 S. Euclid Ave.
St. Louis, MO 63110
Phone: (314) 362-8683
Fax: (314) 362-2692
Robert E. Schmidt, M.D., Ph.D.
Professor of Pathology & Immunology
Department of Pathology and Immunology
Division of Neuropathology
Mailing Address: Campus Box 8118
660 South Euclid Ave.
St. Louis, MO 63110
Office Location: West Building, Room 3720
Office Phone: (314) 362-7429
Lab Phone: (314) 362-7416
Fax: (314) 362-4096
Histopathology and EM
Technical Director – Research EM Facility
Phone: (314) 362-7462
Metabolic and Functional Analysis
Cris Brown (Welling)
Research Lab Supervisor
Phone: (314) 362-8684
Processing and paraffin embedding of fixed tissues or cell pellets. Also available large specimen whole mounting.
5 µm thick slides are cut from paraffin embedded or frozen tissues.
Standard hematoxylin and eosin (H&E) staining for tissue architecture and cellularity as well as 26 special stains are offered for slides from paraffin embedded tissue sections and in some instances on frozen sections.
Investigators and trainees can be instructed to use a cryostat and then elect to cut their own sections.
The image is an H&E section of a normal human islet which consists of a rounded collection of endocrine cells with clear cytoplasm and round nuclei with clumped chromatin. These sit amidst the acini of the exocrine pancreas which has cells with mixed basophilic and acidophilic cytoplasm and scattered adipocytes which are large, round, and completely clear because the lipid material is flushed out with tissue processing.
Routine tissue EM
Processing and embedding (maximum of 6 blocks), cutting 1 micron thick plastic sections stained with toluidine blue, cutting of thin sections on standard 200 mesh grids, post-staining in uranyl acetate and lead citrate, and consultation.
Tissue culture EM
Service includes routine procedures described above but requires additional specimen preparation and cutting time.
Services include LR White embedding media or pre-embed immunostaining, processing and embedding (maximum of 6 blocks) or cutting 1 micron plastic sections on slides – stained for toluidine blue. Additional services include cutting thin sections on formvar grids, primary antibody staining (primary antibody supplied by investigator), secondary antibody gold labeling (secondary antibody charged to investigator), counterstaining with OsO4, uranyl acetate, lead citrate, and consultation.
Services include preparation of formvar coated grids and negative staining techniques appropriate for the virus or particle to be visualized.
This technique produces 3D images of cells on the basis of multiple 2D projection images of a 3D object, obtained from several orientations by tilting the specimen in its holder in the microscope over a wide range of viewing directions.
Training and Consultation, Image aquisition
Consultation includes experimental planning, technical advice on tissue fixation, and discussion of appropriate ultrastructural analysis.
Metabolic and Functional Analysis
Quantification of Metabolic Rates in Diabetes-Related Tissues and Cells
Seahorse XF Extracellular Flux Analyzers, in a 24 well format (XF24-3) and a 96 well format (XF96) are available for study of diabetes and metabolism with the capacity to quantify cellular respiration and glycolysis and the real time kinetics of bioenergetic pathways. Core staff provides training for instrument and software use and the Core director assists the users in the design of their bioenergetics experiment and analysis of their data. Different levels of service are provided. With Premium Service Level, the users only provide the objective of the experiment and the cells or tissues in the XF plates. The Core staff performs the experiment and the data analysis. With Regular and Basic Service Levels, the users perform all steps from experimental design to data analysis and consumables are supplied by the Core or by the user, respectively. XFXF Analyzer consumables, such as microplates for cells, XF reagents, and stress test kits are available in the Tissue Culture Support Center for purchase.
Proliferation, Differentiation, Cell Death and Regulatory Signaling in Diabetes-Related Tissues and Cells
Expertise, equipment, and training for assessment of the effects of nutrients, growth factors and other compounds on growth, proliferation, metabolism, apoptosis, autophagy and differentiation of pancreatic islets and other diabetes-related tissues. Validated antibodies for specialized analysis of pancreatic islets and other diabetes-related tissues and protocols for their application are available.
|Currently stocked antibodies for IHC and/or IF (human, mouse or rat)|
|PCNA||4E-BP sampler kit||phospho-CREB
Human and Rodent Islet Specific Protocols
The core provides established protocols, reagents and instruction for culture and assays of insulin secretion from human and rodent islets by static incubation or perifusion to distinguish first and second phase insulin secretion. In addition, the core maintains stocks of b-cell-derived established lines (Min6, RINm5F, INS-1). The Core assists users in acquiring purified human islets for basic research through the Integrated Islet Distribution Program (IIDP) sponsored by the National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK).
|Core Service||Cost to DRC members|
|Paraffin processing & embedding||$3.75 per block|
|Paraffin sections unstained||$2.25 per slide|
|Paraffin sections H&E||$5.25 per slide|
|Paraffin sections special stains||$6.75 per slide|
|Frozen sections unstained||$6.00 per slide|
|Frozen sections H&E||$7.00 per slide|
|Frozen sections trichrome or PAS||$8.00 per slide|
|Tissue Culture EM||$318.00|
|EM Microscope Time||$65.00/h|
|Seahorse XF Extracellular Analyzer|
|Immunocytochemical Staining Services|
|Training and initial supplies||No Charge|
|Microscopy and Digital Image Acquisition||No Charge|
|Training in islets for functional assays||No Charge|
|Consultation for experimental design, data analysis and grant proposal development||No Charge|