Core Facilities

Researchers at the facility will have access to the Biomolecular Core for molecular genetics research. The Biomolecular Core Laboratory is a centralized, state-of-the-art facility that offers expertise in molecular genetics and genomics and is directed by Dr. Katia Sol-Church. Services offered by the Biomolecular Core include automated DNA sequencing, DNA fragment analysis, pyrosequencing, and custom oligonucleotides from IDT can be purchased through the facility at a discounted price.

Additionally, it is home to the ABI 7900HT Fast Real-Time PCR system containing a 384 well-plate block to allow for unprecedented throughput. The Biomek 2000 can perform automatic liquid handling in a high-throughput manner to complement the capabilities of this system.

For more information, please visit http://www.nemoursresearch.org/cores/bcl/services.html.

The NCCCR members have access to the Cell Science and Proteomic Core directed by Dr. Robert Mason. The Cell Science Core is equipped with Class II culture hoods (2), incubators (6), fluorescent and phase contrast microscopes, and associated cell preparative facilities. Confocal microscopic facilities are available in an adjacent laboratory. A Beckman Coulter Epics Altra FACS is available in the nearby Research and Administration building for fluorescent sorting of cells in a sterile environment. The front-end proteomics facility is equipped with large-format 2D gel electrophoresis (12 plate capacity) and several small-format gel systems. A trained staff member is available for spot-picking, protein-digestion and MALDI plate spotting. A Typhoon phosphorimager and associated software are available for analysis of DIGE labeled 2D gels and other fluorescent samples. Expertise is also available for HPLC analysis.

The NCCCR members have access to the Coulter EPICS XL-MCL Flow Cytometer within the Research Immunology laboratory, under the direction of Dr. Paul Fawcett, provides versatile flow-cytometric analysis for the research setting. This system has the capability to analyze up to 4 colors of immunofluorescence from the uniphase 488nm Argon ion laser, which eliminates concerns regarding multi-beam stability, signal delay and alignment. The high performance photomultipliers provide spectral sensitivity from 200nm-800nm for fluorescence detection. Multi-color applications are available for cell surface marker analysis and DNA analysis, among others. The Digital Signal Processing (DSP) provides reliable linearity and drift-free amplification and compensation. In addition, the EXPO32 ADC software allows for customized acquisition and analysis options to enable researchers to get the most information out of their data.

Histology Core facility: A full fledged histology lab with facilities for tissue fixation, processing for frozen and paraffin embedded tissues is available to NCCCR members. For more details please visit this website.

Biomedical Analysis Laboratory (BAL), Nemours Children’s Clinic (NCC) Jacksonville, FL

BAL occupies approximately 2200 sq. ft. in the clinic building and is equipped with various state-of-the-art scientific instruments.

1. Mass spectrometry: Research using stable isotope dilution and mass spectrometry-based techniques is one of the major areas of focus of the lab for analytical and clinical research to follow various molecules in the human body and study their metabolism. Stable isotopes are not radioactive and they are present in nature in small quantities. Their use in humans is perfectly safe and studies using stable isotopes have been performed even in children and infants, including preterm neonates. Overall stable isotope dilution and mass spectrometric studies provide better insights into the underlying mechanisms of alterations in amino acid, protein, carbohydrate and energy metabolism in a disease condition and/or in response to an intervention. We use gas chromatography-mass spectrometry (GCMS) and isotope ratio mass spectrometry (IRMS) instrumentation. Currently the laboratory has four mass spectrometers: two GCMS (HP (Agilent) 5973 capillary GCMS) with electron impact (EI), positive chemical ionization (PCI) and negative chemical ionization (NCI) detectors and two IRMS – one VG Optima Isochrome Model SIRA 3 magnetic sector GC-combustion-IRMS (GC-C-IRMS) and one PDZ-Europa (Cheshire, England) GC-combustion/pyrolysis-IRMS (GC-C/P-IRMS). These instruments are mainly used for the measurement of concentration of different molecules as well as turnover of amino acids and proteins using stable isotopes of oxygen, hydrogen, nitrogen and carbon. These techniques are highly useful in studying various disease conditions such as obesity, CVD, diabetes and cancer.
2. Other major instruments: Complementary to the mass spectrometric techniques the BAL is equipped to do a variety of measurements using a range of instrumentation such as HPLC, electrophoresis, affinity chromatography and nephalometry in addition to various Radio-Immuno Assays (RIA) and Enzyme-Linked Immuno-Sorbent Assay (ELISA) for hormones, peptides and proteins. List of other major instruments:

1. Nephalometer (BN II Nephelometer, Dade-Behring): This instrument is mainly for the high-sensitivity measurement of concentrations of an array of plasma proteins such as CRP, fibrinogen, RBP4, transthyretin, Apo A1, ApoB-100 and complement proteins
2. Beckman Model 6300 amino acid analyzer: for the complete analysis of amino acids in physiological fluids and protein hydrolysates; it provides detailed information regarding the relative amino acid composition and free amino acids
3. Bio-Rad Prep Cell system: For the purification of specific muscle proteins such as (myosin heavy chain)
4. Immununo-affinity chromatography system (Bio-Rad): For the purification of various plasma proteins

Small Animal Molecular Imaging Facility at Thomas Jefferson University

The Jefferson Small Animal Imaging facility consists of a Siemens, high sensitivity, high resolution micoroPET scanner “Inveon”, a Siemens microCT ImTek II scanner and Philips micro PET/multi-pinhole SPECT scanner . The Imaging facility now 5 years in service, is available to all investigators on the campus as paid service and is managed by Dr. Kumar, an experienced medical imaging physicist and is directed by Dr. Thakur, Prof. of Radiology. All imaging protocols are discussed with each investigator, for clarity of imaging procedures and for the animal welfare.