Spotlight Stories

Leica CryoJane Tape-Transfer System now available at the Histology Research Core Facility

Annabelle Stein — 2013-04-22T12:57:25Z

Briefly, a cold adhesive tape is applied to the frozen sample, supporting the section as it is cut. The tape, with attached tissue, is rolled onto a cold adhesive-coated slide which is then cured with a flash of ultraviolet light. As a result, the adhesive coating is converted into a solvent-resistant plastic. The tape is then pulled away leaving the section tightly bonded to the slide.

The Tape-Transfer System preserves sample morphology and produces frozen sections with the same quality as those cut from paraffin blocks. Sections are wrinkle-free, uncompressed and fully intact. This system is ideal for projects requiring perfect morphology and/or ultra thin sections that reveal morphological detail and clarify structural relationships. CryoJane applications include histological stains, immunohistochemistry, immunofluorescence, in situ hybridization, laser capture microdissection, frozen tissue arrays and other molecular diagnostic studies.

The Histology Research Core Facility offers a variety of services, insuring that histological techniques are always available. Currently we provide histological and immunohistochemical services to users from UNC campuses and other research institutions. The Facility is located in Rooms 004 and 007 of the Glaxo Research Building. For further information please contact Kirk McNaughton, Histology Research Core Facility Director, at macone@med.unc.edu (919) 966-1202 and check out our web page www.med.unc.edu/physiolo/research/histology-facility/
.

High-Throughput Peptide Synthesis and Array Facility Assists UNC scientists link histone methylation with DNA methylation in humans

Annabelle Stein — 2012-11-19T14:20:00Z

The study found that UHRF1, an E3 ubiquitin ligase, regulates DNA methylation through its association with histone H3 methylated at lysine 9 (H3K9). Using advanced peptide arrays technology featured in the HTPSA Core Facility, Dr. Strahl’s group found that UHRF1 association with H3K9 methylation can occur during mitosis – a cell cycle stage wherein other H3K9 methylation binders are normally ejected. The collective studies showed that UHRF1 mitotic binding to chromatin is necessary for DNA methylation maintenance through regulation of the stability of the enzyme that methylates DNA. This work is significant because it connects the regulation of two fundamentally important epiegentic marks together in humans. Furthermore, it shows how peptides and peptide arrays can be a useful tool for the investigation of biologically important protein-protein interactions.

The HTPSA Core Facility helped this study by providing high quality peptides and peptide arrays. The facility can help in design and synthesis of peptides for antibody production, Fluorescence Polarization assays, NMR and crystallography. The core has experience in fast synthesis of simple peptides and synthesis of peptides containing multiple tags and/or modifications, including phosphopeptides, fluorescent peptides, biotinylated peptides, peptides with post-translational modifications, and peptides with unnatural amino acids.

If you thinking about using peptides in your research, contact the HTPSA Core to discuss the optimal peptide design, advantages and disadvantages of using peptides, and answer your questions about peptide stability, handling, and storage. For more information, visit the HTPSA Core Facility Website or email us at peptides@unc.edu

Translational Research Now Possible Through the Human Pluripotent Stem Cell Core

Annabelle Stein — 2012-06-26T15:25:00Z

The reprogramming technology became available a few years ago when scientists proved that a somatic cell could be de-differentiated to a stem cell state. The vision and creativity of this technology is emphasized by its potential use in personalized therapy; induced pluripotent stem (iPS) cells represent a new and exciting avenue to generate patient-specific stem cells. Moreover, because iPS cells are made from a patient’s own cells the difficulties associated with tissue rejection are eliminated. Importantly, iPS cells derived from patients with various diseases represent an invaluable research tool to study the mechanism of diseases and to identify new therapies.

HPSCC is equipped to generate iPS cells from a variety of somatic cell types using lentiviral transduction however integration-free reprogramming will soon be a service offered by the Core. Generation of integration-free iPS cells will allow for their direct use in translational applications. Derivation of hiPS cells is just one of the resources that the HPSC Core offers. The study of the human ES cells offers an invaluable insight into the early human development and the unprecedented opportunity to study the molecular mechanisms underlying pluripotency and differentiation. Thus, hES cells represent a potentially unlimited source of cells for cell replacement therapies as well as drug screening studies. HPSC Core has available several federally approved human ES cell lines. Protocols have been established in the HPSC Core to differentiate both hiPS and hES cells into a variety of cell types. Housed in Taylor Hall, the hPSC Core is equipped with all the reagents necessary to generate iPS cells, to expand hES cells as well as for differentiation studies. Please visit the Human Pluripotent Stem Cell Core website for more information.

Cutting-edge Metabolic and Cardiorespiratory Testing Equipment Now Available Through the Nutrition Research and Metabolism Core

Annabelle Stein — 2012-04-26T20:05:00Z

The Nutrition Research and Metabolism Core has upgraded their metabolic cart to a state-of-the-art MedgraphicsUltima Series CPX metabolic cart. The Ultima indirect calorimeter calculates resting energy expenditure and substrate utilization through breath by breath analysis of O2 and CO2. In addition to the metabolic testing capabilities, the Ultima is used for multiple tests of pulmonary function and in exercise testing. As the name suggests, the Ultima CPX sits on a mobile cart that can be used in any room of the Clinical Translational Research Center or other locations within the hospital.

Indirect calorimetry is just one of the resources the Nutrition Research and Metabolism Core offers to investigators. Many obesity research protocols measure resting metabolic rate in conjunction with body composition using their Hologic Bone Densitometer. In addition, the Core’s dietitian and fully staffed metabolic research kitchen implement feeding studies to control any nutrient of interest. In collaboration with the Clinical and Translational Research Center nursing staff, they are able to implement nutrition research protocols of any intensity.

BioBanking: There are four major campus facilities that process and bank human specimens

Annabelle Stein — 2012-03-01T13:45:00Z

These facilities may provide additional services such as extraction and storage of DNA or RNA, and advice about preanalytic study design (specimen collection, assessing specimen quality).
The NC TraCS Biobanking Consortium coordinates services to help investigators collect, process, and store human specimens for research.
Click here for more information

Versatile Multiphoton Confocal Microscope System at the Michael Hooker Microscopy Facility

Annabelle Stein — 2011-11-14T13:50:00Z

The system is an Olympus FV1000 MPE SIM on an inverted microscope stand and features an environmental enclosure with temperature, gas, humidity control and light exclusion. An extensive range of standard lasers are available: 405 nm for UV dyes (e.g. DAPI), 440 nm for eCFP, 488 nm for e.g. FITC, 514 nm for e.g. YFP, 559 nm for e.g. Rhodamine, Texas red/dsRed & 635 nm for e.g. CY5/Alexa 633. These lasers also permit imaging of standard and more exotic FRET pairs. Scanning in x-y & z axes allows serial section image collection for later 3-D rendering and analysis. The motorized x-y stage allows multiple area time lapse experiments and tiling/montaging in 2-D & 3-dimensions or the imaging of multiwell micro-titer plates. The SIM module can do rapid photo-activation or fluorescence recovery after photobleaching protocols.

Mutiphoton confocal imaging is powered by a pulsed IR laser tunable from 690 to 1040 nm. Deep tissue imaging is enhanced by pulse precompensation. Standard spectral or high sensitivity non-descanned detectors provide added flexibility.

In support of this versatile resource a 37C, 5% CO2, 95% humidity an incubator is available for sample storage, and network accessible raid disk array long term image storage. Off line imaging software include the Olympus offline viewer and Volocity volume visualization software, with license server (http://microscopy.unc.edu/volocity/), for 3D and time lapse rendering, analysis, movie production, 3-D particle tracking, object counting or co-localization analysis and deconvolution.

The system is funded by the North Carolina Biotechnology Center and the School of Medicine. Users from the UNC campuses and other research institutions are welcome. Please contact the MH Microscopy Facility personnel for demonstrations, instruction, requests and more information.

The Facility is located in the Thurston Bowles Building, room 6129.

Innovative Human Imaging System coming soon to UNC

Annabelle Stein — 2011-09-29T18:00:00Z

This new system represents a major technological breakthrough in the imaging field. Specifically, this system allows simultaneous acquisition of MR and PET images in human subjects. "Simultaneous" is emphasized since it allows acquisition of both MR and PET images at the same time without delay. For a comparison, current PET/CT acquires PET and CT images in a sequential fashion, acquiring PET images, followed by moving patient table to the CT end for obtaining CT images. Therefore, combining the detailed anatomical information offered by MR, high sensitivity to in vivo molecular events from PET, and substantially reduced iodize radiation when compared to PET/CT, this new MRPET scanner most likely will not only improve patient throughput but also open new avenues of research projects and clinical applications.

We are very fortunate to receive the fourth whole body MRPET scanner in the US (behind MGH, NIH, and WU), which should give us tremendous advantages in new grant funding opportunities as well as be the leaders of this new field of imaging research.
The BRIC will host a MR/PET symposium to kick off this exciting endeavor. The symposium is scheduled for Thursday, November 10, 2011. Please see Events section for more information and link to registration.

Biomarker Mass Spectrometry Facility Contributes to Discovery

Annabelle Stein — 2011-08-31T12:52:26Z

With a finding published online in the July 21, 2011 issue of the journal Science, researchers from the University of North Carolina at Chapel Hill School of Medicine and Gillings School of Global Public Health have discovered the seventh and eighth bases of DNA. These last two bases – called 5-formylcytosine and 5-carboxylcytosine – are actually versions of cytosine that have been modified by Tet proteins, molecular entities thought to play a role in DNA demethylation and stem cell reprogramming. The discovery could advance stem cell research by giving a glimpse into the DNA changes (such as the removal of chemical groups through demethylation) that could reprogram adult cells to make them act like stem cells.

"Before we can grasp the magnitude of this discovery, we have to figure out the function of these new bases," said senior study author Dr. Yi Zhang, Kenan Distinguished Professor of Biochemistry and Biophysics at UNC and an investigator of the Howard Hughes Medical Institute at the Lineberger Comprehensive Cancer Center. "Because these bases represent an intermediate state in the demethylation process, they could be important for cell reprogramming and cancer, both of which involve DNA demethylation." Much is known about the "fifth base," 5-methylcytosine, which arises when a chemical tag or methyl group is tacked onto a cytosine. This methylation is associated with gene silencing, as it causes the DNA's double helix to fold even tighter upon itself. Last year, Dr. Zhang's group reported that Tet proteins can convert 5 methylC (the fifth base) to 5 hydroxymethylC (the sixth base) in the first of a four-step reaction leading back to bare-boned cytosine. But try as they might, the researchers could not continue the reaction on to the seventh and eighth bases, called 5-formylC and 5-carboxyC. The problem, they eventually found, was not that Tet wasn't taking that second and third step, it was that their experimental assay was not sensitive enough to detect it. Once they realized the limitations of the assay, they redesigned it and were able to detect the two newest bases of DNA with electrophoresis, but reviewers wanted more detailed chemical analyses. The Zang laboratory contacted Dr. James Swenberg, Kenan Professor of Environmental Sciences and Engineering, who introduced them to Leonard Collins from the Biomarker Mass Spectrometry Facility Core. Together, they quickly developed advanced mass spectrometry methods to fully characterize the new bases and demonstrated the presence of as little as 5 fmol of the minor bases in stem cells and mouse tissue DNA. Other study co-authors include UNC Lineberger Comprehensive Cancer Center postdoctoral research associates Dr. Shinsuke Ito, Dr. Li Shen and Dr. Susan C. Wu.