SC BioCRAFT

Cores

Bioengineering and Bioimaging Core

Biomaterials Characterization: 
Dr. Robert Latour, Core Director, Supervisor
Dr. Gulya Korneva(gkornev@clemson.edu), Lab Support, 864-656-5134

Bioactive Matrix Synthesis and Bioreactor Design:
Dr. Ken Webb, Supervisor
Dr. Gulya Korneva, Lab Support

Biomechanics:
Dr. Jiro Nagatomi, Supervisor
Dr. Gulya Korneva, Lab Support

Bioimaging:
Dr. John Parrish, Supervisor
Dr. Ann-Marie Broome, MUSC, Lab Support

The overarching theme of SC BioCRAFT is regenerative medicine, a highly interdisciplinary field; expertise and support in bioengineering methods as well as cell and molecular biology are required to succeed. Junior investigators in our IDeA state need core support to further their research and compete nationally for grant funding. From our experience in Phase I and from several discussions with our internal advisory committee and targeted PIs, we have identified four bioengineering thrust areas that PIs will need core support in. To meet these needs, the previously named “Materials Synthesis, Characterization, and Testing Core” has been expanded and renamed the “Bioengineering and Bioimaging Core (B5)”. This is the only bioengineering core in the COBRE programs throughout the US (out of > 200 cores). This Core will provide support for both COBRE and outside investigators for their research needs under the areas of Biomaterials characterization, Bioactive matrix synthesis and Bioreactor design, Biomechanics, and in vivo Bioimaging. The core will maintain a broad range of instruments and provide technical support and training for facilities for each of these areas.

Clemson University has significantly invested in developing this core in Phase I, including the addition of new laboratory space and advance instruments at Clemson’s main campus and at Clemson University Biomedical Engineering Innovation Campus (CUBEInC) in nearby Greenville, SC, and addition of faculty with specific expertise in different thrust areas of this core that include targeted investigators in Phase I.

The overall goal of the Bioengineering and Bioimaging Core is to provide resources, facilities, and training to support research related to tissue engineering, regenerative medicine, and drug delivery-system design for designated COBRE PIs and other research groups within and outside the State of South Carolina to increase the state’s competiveness in biomedical research. This goal will be achieved through four specific aims:

  • Aim 1: To provide facilities and technical support for biomaterial and biointerface characterization
  • Aim 2: To provide technical support for bioactive matrix synthesis and bioreactor design
  • Aim 3: To provide facilities and technical support for biomechanical characterization and testing
  • Aim 4: To provide facilities and technical support for in vivo bioimaging

Instruments at the BB Lab:

  1. Spectropolarimeter (Jasco J-810)
  2. Temperature Controlled Automatic Refractometer AR700 (Reichert)
  3. Contact angle goniometer DSA-20E (Krüss)
  4. Variable-angle Spectroscopic Ellipsometer GES 5 (Sopra Inc)
  5. Flame Atomic Absorption - AAnalyst 200 Instrument (Perkin Elmer)
  6. Surface Plasmon Resonance Spectrometer Biacore X
  7. Materials Testing System (MTS) Synergie 100
  8. HPLC/GPC System (Waters)
  9. Expanded Plasma Surface Treatment Unit PDC-001 (Harrick Plasma)

 

Spectropolarimeter (Jasco J-810)
Measures the circular dichroism (CD) and optical rotary dispersion (ORD) of the sample.
Specifications:
Wavelength range from 163 to 900 nm
Accessories: Peltier type temperature controller Jasco PFD-425S; Automated pH titrator Jasco ATS-429S.
Applications:

  • Allows to verify the chirality of molecules or compounds;
  • Using available software, fractions of each different conformation in protein structure can be estimated;
  • The conformational changes of molecules with temperature can be studied;
  • Useful for verifying that the protein is in its native conformation;
  • Protein folding studies; and
  • Purity testing of optically active substances.

 

Spectropolarimeter
Temperature Controlled Automatic Refractometer AR700 (Reichert)
Specifications: Reading range of 1.33000 to 1.60000 Refractive Index and the full Brix scale. Precision: to 0.01 Brix or 0.00001 RI
Applications: Measures refractive index of transparent, translucent, and opaque liquids.
Refractometer
Contact angle goniometer DSA-20E (Krüss)
Applications:
  • Determination of static and dynamic contact angle;
  • Determination of surface and interfacial tension of liquids; and
  • Determination of automatically measured surface free energy of solids.
DSA20E
Variable-angle Spectroscopic Ellipsometer GES 5 (Sopra Inc)
Features: Thickness measured: from monoatomic to several microns thick. The beam size: 1 mm2 and 10 mm2; The variable angles: from 7º to 90º with variable +/- 0.01º; spectral range from 200 nm to 800 nm
Applications:
  • Measures material refractive indices;
  • Layer thicknesses of thin film layers on flat substrates; and
  • Up to 7 layers can be analyzed in terms of thickness and optical characteristics for each layer.
Ellipsometer
Flame Atomic Absorption Spectrophotometer - AAnalyst 200 Instrument (Perkin Elmer)
Specifications: Wavelength range from 189 to 900 nm.
Applications: For determining very precisely the concentration of a particular element in a liquid sample. Can detect from 1 to 1000 µg/L. For each element, the specific lamp is required. Ca, Cu, and Fe lamps are available. The list of lamps/elements for analysis on AAnalyst 200 can be found here: VWR
AAnalyst 200
Surface Plasmon Resonance Spectrometer Biacore X
Biacore is a methodology for measuring the interactions mostly between proteins, in real time using the phenomenon of surface Plasmon resonance (SPR).
Specifications:
  • Two Flow cell capability;
  • Temperature regulation from 4 to 40oC;
  • Flow rates 1-100μL/min, steps of 1 μL;
  • Required sample volume: injection volume (minimum 5 μL +20 μl);
  • Molecular weight detection: >180 Da;
  • Refractive index range from 1.33 to 1.40; and
  • Sensitivity: 70000 RU*, where RU, resonance units, 1RU = 1 pg/mm2 of sensor surface.
Applications:
  • Kinetics of binding;
  • Solution affinity;
  • Steady state affinity;
  • Yes/No binding;
  • Ligand fishing;
  • Affinity ranking;
  • Protein concentration assays;
  • Protein percent activity assays; and
  • Thermodynamics.
Interactions between nucleic acids, lipids, small molecules, and whole cells can be studied.
Spectrometer
Materials Testing System (MTS) Synergie 100
Applications: Tensile, Flex, Peel/Tear, Strain, and Compression test methods.
Specifications:
  • Loadings available are 10 N (2 lbf) and 100 N (22.5 lbf);
  • Speed range from 0.001 to 1000 mm/min (0.00004-39 in/min);
  • Max specimen thickness for grips: 13 mm (0.5 in); and 
  • 50 mm (2.0 inch) diameter lightweight compression platens. Face set, 25x25 mm rubber, for 100/2000 advantage pneumatic and screw action grips.
MTS
HPLC/GPC System (Waters)
Applications: MW determination, high resolution separation and purification.
Features: Binary Gradient Pumping System, Autosampler, Photodiode Array Detector (Wavelength Range from 190 to 800 nm with 1.2 nm resolution with wavelength accuracy: ± 1 nm); Refractive Index Detector (Refractive Index Range: 1.00 to 1.75 RIU with temperature control from 30 to 55oC); Multi Lambda Fluorescence Detector (Operates from 200 to 900  nm); Column Heater (Temperature Range from 20oC to 60oC; with temperature accuracy of ±0.8oC; Column Capacity of up to four 7.8 mm x 300 mm columns); Fraction Collector (Methods of Operation: time, drop count, drop volume, signal).
HPLC
Expanded Plasma Surface Treatment Unit PDC-001 (Harrick Plasma)
Plasma Applications
Features:
  • Expanded Pyrex plasma chamber 6″diameter x 6.5″ length (15.24cm x16.51 cm);
  • PlasmaFlo gas mixer; and
  • Oxygen service pump.
PDC

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Cell, Tissue, and Molecular Analysis Core

Molecular Biology:
Dr. Rick Visconti
Dr. Dan Simionescu
Dr. Shibnath Ghatak

Histology and Advanced Imaging:
Dr. Terri Bruce, Co-Director
Linda Jenkins
Chad McMahan
Dr. Bruce Gao

Stem Cell Biology:
Dr. Carol Brenner, Co-Director
Dr. Dan Simionescu

Cell, tissue, and molecular biology techniques are the cornerstone of biomedical research and regenerative medicine endeavors. A major obstacle for junior faculty is lack of state-of-the-art facilities and specialized expertise commonly found in states with major NIH funding. This often hinders the generation of preliminary data necessary to be competitive for NIH R01 grants. In the State of South Carolina, there are several strong facilities and successful investigators who have received generous NIH funding and have gained significant tissue, cell, and molecular expertise. However, junior faculty in SC have not benefited optimally from these opportunities. To close this gap, we developed the Cell, Tissue and Molecular Analysis Core (CTMA core) to be affiliated with SC BioCRAFT.

After conferring with the PI of the COBRE proposal and the five targeted PIs, we have identified three thrusts that will serve the needs of all our COBRE projects: 1) molecular biology, 2) histology and advanced imaging, and 3) stem cell biology. Outstanding facilities and labs directed by expert investigators in these three thrusts have been identified and brought together to serve the needs of COBRE PIs and those of other investigators in the State of South Carolina in CTMA core. Areas of expertise in this core include regenerative medicine, gene and protein expression and manipulation, adult and induced stem cells, and advanced histology, electron microscopy, and biophotonics. The Core will provide support to develop ideas, overcome technical challenges, and lead the targeted projects towards significant outcomes as outlined in the following specific aims:

  • Specific Aim 1 – To promote and facilitate advanced molecular biology research
    Two main research facilities, the Regenerative Medicine Laboratory in Clemson and the Signal Transduction Facility at MUSC, are fully functional and will provide unlimited expertise with gene and protein expression, silencing, transfection, traditional biochemistry assays, signal transduction, and targeting nanoparticles. Additional expertise and services in cell sorting, microarrays, molecular engineering, proteogenomics and bioinformatics are available from collaborating P30 MUSC COBRE cores. Together, these facilities, operated and supported by Clemson University and MUSC, are fully equipped, have impressive potential, and are ready to serve the SC COBRE investigators and others throughout the state.
  • Specific Aim 2 – To provide histology and advanced imaging expertise and support
    Many of the research questions posed by the COBRE investigators will require histology and advanced microscopy to answer. The ability to visualize the subcellular locations of biomolecules and organelles will be crucial to these researchers as they strive to engineer translatable solutions to challenging tissue engineering problems. The histology and microscopy facilities will provide the tools investigators need and will foster a collaborative environment that introduce the newest techniques in histology and advanced imaging.
  • Specific Aim 3 – To enhance adult and induced stem cell research capabilities
    Stem cell research facilities, such as the Laboratory for Regenerative Medicine and the Stem Cell Institute, will provide expertise with derivation, characterization and differentiation of induced pluripotent stem cells and adult stem cells, and in vivo tracking, in collaboration with the Bioengineering Core. Since stem cells are such an important tool for regenerative medicine, all targeted investigators will be encouraged to learn about them and consider ways to employ them in future grant applications. The laboratory personnel are highly qualified in the field, and the labs are fully equipped and functional and ready to help the targeted PIs. The Stem Cell Institute and Clemson University, respectively, operate, support and maintain the core facilities. The proposed core will greatly enhance research capabilities and most significantly advance the potential for regenerative medicine research within South Carolina. While COBRE trainees will be the primary beneficiaries and priority users of this facility at no cost, this core will also support the research programs of faculty members at collaborating institutions throughout the entire state.

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