Infrastructure SRP Diabetes
Metabolic Phenotyping Center for Diabetic Animal Models
A centralized metabolic and physiologic phenotyping core center for genetically modified diabetic mouse models specifically focused on diabetes and its complications has been established. Standardized protocols for animal handling procedures, metabolic screens (including glucose and insulin tolerance tests), blood chemistry analysis (Luminex), meal tolerance testing, body composition (EchoMRI analysis), whole body energy homeostasis (Comprehensive Lab Animal Monitoring System; CLAMS), whole body glucose homeostasis (euglycaemic-hyperinsulinaemic clamp technique in conscious mice), exercise intervention (running wheels), and behaviour. The aim is to provide not only the facility, but also advice on protocol design and data interpretation. Interest in using the platform has been high, and we are currently planning a major investment (in the range of 3 million SEK) to expand capacity for the metabolic cages.
Presentation of the facility:
Center for Clinical Metabolic Research in Diabetes
The center is a unit for outpatient investigations of human subjects using established as well as novel types of techniques. The center has recently moved to new facilities in the C2-building of the Karolinska University Hospital in Huddinge where it is integrated with the outpatient clinic of the Endocrinology and Diabetology unit. This ensures a close interaction with the clinical activities and access to diabetic subjects for clinical studies. At the moment the center has eight affiliated senior medical doctors performing the clinical studies and employs one research nurse and one technician (both full-time). Starting this year, we will employ an additional full-time research nurse. In addition, the center has the possibility to involve three additional research nurses employed at the research unit of our clinic, this has been a great advantage given the large clinical studies which have been initiated during the last year. Taken together, the available facilities and personnel enables the center to offer a wide range of clinical assessments including: clamps, microdialysis of WAT and muscle, isotope turnover studies, assessments of pharmacodynamics-kinetics, indirect calorimetry, body composition (bioimpedance and DXA), tissue biopsies (WAT and muscle) and evaluations of arterial stiffness (Arteriograph). In addition, we can also perform 24 hr-monitoring studies.
Presentation of the Center
Beta Cell Imaging
A facility at the Rolf Luft Center (Department of Molecular Medicine and Surgery) with state-of-the art microscope technology for visualizing beta cells in different experimental settings is supported by the programme. The facility could also be used for imaging other cell-types. Instrumentation for in vitro fluorescence-based microscopy includes one confocal microscope, one TIRF (total internal reflection fluorescence) and two microscopes for dynamic microfluorimetry. High-through put imaging can be performed using high content screening (BD Pathway 855 instrument) as well as fluorescence activated cell sorting (FACS with 5 lasers and 16 detection channels) for analyses and sorting of cells. Two upright microscopes are equipped for in vivo imaging using confocal and multiphoton laser-scanning microscopy. A unique platform has been developed at the facility to enable non-invasive studies of islets transplanted to the anterior chamber of the eye (ACE, mouse or rat) where the cornea serves as a natural ‘body-window’ for in vivo imaging.
SciLifeLab: Technology platforms available at this center for high through-put molecular bioscience in Stockholm will be specifically tailored to adress the diabetes focused research in this programme. Bioinformatic support is available on a collaborative basis for researchers in SRP Diabetes.
BEA: The Bioinformatics Expression Analysis Core Facility, affiliated to SciLifeLab, located at Novum Research Park at Huddinge Campus have experience in obesity and metabolism research and can offer paid service or help on a collaborative basis. It contains the following technology platforms: Affymetrix, both the classical and high throughput (96 well) format; Agilent; Applied Biosystems platform for real-time PCR, including 96, 384 and TLDA formats; Illumina, including the array platform and access to the high throughput sequencing platform. The infrastructure also includes robots for RNA purification and a repertoire of platforms for RNA and DNA quality control and quantification.
Applications include transcript profiling (including mRNA, miRNA and total transcript profiling); genetic analysis (including genome wide association studies (GWAS) and copy number analysis); epigenetic analysis (including DNA-methylation and histone modifications); global DNA-binding by transcription factors; bioinformatics support for all these applications. Recently, proteomics using mass spec was established in connection to the Dahlman-Wright group and is also available for the SRP Diabetes consortium.
Presentation of the BEA facility
Contact Person BEA facility