Technologies

Research Tools

Most Recent Inventions

Efficient In Vitro Assay for Antigen-Specific Tolerance

Building on their work, UW–Madison researchers have now developed a T cell-bound cytokine (T-CBC) assay for detecting and quantifying regulatory T cells specific to self-antigens or donor alloantigens. The new method comprises (a) culturing the subject’s T cells for 24 hours in the presence of one or more target antigens and (b) analyzing the cultured T cells for expression of a marker (EBi3; TGFβ/LAP) indicative of antigen-specific immune suppression.
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Monomeric Fluorescent Protein-Ligand Complexes with Strong Fluorescence in the Far-Red Region

Research from the University of Wisconsin-Washington County in collaboration with the Institute for Stem Cell Biology and Regenerative Medicine in India, has resulted in the development of monomeric variants of the naturally occurring Sandercyanin Fluorescent Protein (SFP) using site-directed mutagenesis. This work has stemmed from earlier research focused on development of the tetrameric form of SFP, a biliverdin-binding lipocalin protein originally isolated from the mucus of the blue walleye fish, Sander vitreus. Monomeric variants of SFP (mSFPs) have been found to possess the same non-covalent, bili-binding characteristics of the tetramer but are one-quarter the size (~18.6kDa) and do not oligomerize. They are therefore anticipated to be more useful in a host of biotechnology applications. Like the tetrameric form, the mSFPs have a large stokes shift (375nm/675nm) and fluoresce in the far-red or near infrared region, which is advantageous for a wide range of applications including investigation of protein-protein interactions, spatial and temporal gene expression, assessing cell biology distribution and mobility, studying protein activity and protein interactions in vivo, as well as cancer research, immunology, and stem cell research and sub-cellular localization. In addition, the newly developed mSFP’s far-red fluorescence is particularly advantageous for in vivo, deep-tissue imaging.
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Enhanced Endotoxin Detection: New Advantages in Liquid Crystal Assays for Gram-Negative Pathogens

UW–Madison researchers have now demonstrated enhanced endotoxin detection in the presence of masking agents in their previous liquid crystal system.

Unlike the LAL assay, the LC-based method does not suffer from LER or any loss of sensitivity due to the presence of cations (e.g., Ca2+ or Mg2+), buffers (e.g., citrate), surfactants (e.g., SDS), chelating agents (e.g., EDTA), proteins or nucleic acids (e.g., DNA or RNA). Thus, the LC-based method provides faster and cheaper detection of endotoxin when compared to existing methods, such as the LAL assay.
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Adapted Rhinovirus C for Maximum Virus Yield

Building on their work, the researchers have now developed a mutated RV-C strain that induces strong cytopathic effect and replicates vigorously in the HeLa-E8 cells, yielding more than a log higher level of infectious rhinovirus particles compared to the parental clinical isolate.
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DNA “Millichip” Enables Low-Cost, High Throughput Gene Expression Analysis

UW-Madison researchers have developed a DNA “millichip” designed for low-cost, high throughput gene expression analysis in whole genomes.  The millichips consist of 1,000 to 100,000 different oligonucleotides probes immobilized on small solid support arrays with relatively high density.  The probes, which range from 30 to 100 nucleotides long, occupy separate, known sites in the arrays. 

For example, a maskless array synthesizer (MAS) can be used to synthesize about 800,000 70-mer oligonucleotides on a glass microscope slide.  Then the slide is divided into 96 pieces, each containing about 30,000 of the 70-mer DNA sequences.  These small pieces can be used in any experiment that uses standard DNA chips.

Because the millichips are small, less than 10 cubic centimeters, small volumes of solutions can be used for analysis.  In addition, the small substrate size allows the arrays to be visualized using instrumentation readily available in research laboratories.
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Most Recent Patents

Microtube Scaffold for Sensing and Stimulating Nerve Cell Connections

UW-Madison researchers have developed a method to produce a scaffold system for neurons that permits guided growth or interconnection of neurons and sensing or stimulation of neural activity. The method involves growing nerve cells through doped semiconductor microtubes that act as tunable electrodes for sensing and stimulating nerve cell connections. The tubes allow the growth and interconnection of the neurons to be controlled, and sensors and/or stimulating probes incorporated along the length of the tubes can be used to provide precisely located but spatially separated measurements and stimulation.

The tubes are made of semiconducting thin-film nanomembranes, may vary in length and have diameters ranging from one to 100 microns. Cells are placed near the opening of the tube and preferentially grow through the tube. The microtubes form a coaxial probe around the nerve cell growth, effectively coupling an electrode to the neurons. The tube also acts to protect the neuron from a culture solution that may produce ion leakage, affecting signal propagation and introducing signal noise.
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Regulating Stem Cell Behavior with High Throughput Mineral Coatings

UW–Madison researchers have developed methods of non-viral cell transfection and regulating cell behavior using mineral coatings. The coatings bind polynucleotides and provide a source of calcium and phosphate ions to enhance transfection.

More specifically, a mineral coating is formed by incubating a substrate in a simulated body fluid (SBF). The substrate then is loaded with a polynucleotide (e.g., plasmids, mRNA or proteins), which binds to the coating. Next, a solution of cells is deposited and cultured until a desired level of transfection occurs.
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New Viral Propagation Method Yields Insight Into Childhood Asthma

UW–Madison researchers have developed an efficient, cost-effective method of propagating RV-C. They discovered that human cadherin-related family member 3 (CDHR3) is the receptor for RV-C and allows cell lines normally unsusceptible to HRV-C infection to support virus binding and replication.

To create cell lines capable of efficiently growing RV-C, the researchers modify the host cell so it expresses an effective amount of the CDHR3 receptor. This method enables high-throughput, large-scale production of RV-C, which in turn enables critical basic and applied research regarding this understudied pathogen.
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