Research Tools & Reagents

Blood transfusions provide important medical benefits primarily centered on the replacement of lost blood in trauma and surgical patients.

However, the availability of blood transfusions in both hospital and field settings is limited by the safety and availability of the donated blood supply. Therefore, blood substitutes have long been sought as one way to alleviate these safety and availability concerns.

To date, current blood substitutes have met with modest clinical success against a backdrop of varied concerns regarding cardiac function, renal toxicity, and molecular stability. Recombinant oxygen-binding heme proteins incorporating circularly permuted globins address these issues through the promotion of cross-linking and the formation of hemoglobin multimers.

Recombinant DNA sequences coding for the hemoglobin protein have been modified to introduce new peptide linkers and termini. The resulting genetic cross-linking and covalent binding promote stability within individual hemoglobin molecules and the formation of high-weight hemoglobin multimers.

Applications

• As a blood substitute in the treatment of traumatic injury in both hospital and field settings

Benefits

• Increased safety and availability of blood substitutes for transfusion
• Potential for increased stability and reduced toxicity in a recombinant protein due to the modifications that promote subunit cross-linking and the formation of hemoglobin multimers

Current super-resolution microscopy techniques require special laser setups and fluorescent molecules (such as PALM and STORM) that allow the fluorophores to be turned on and off so that only a small number of fluorophores are turned on at any time. With the Super Shrimp image analysis algorithm, super-resolution images can be created from images with high densities of fluorophores. The invention creates microscopy images that have 5 or more times better resolution than standard microscopy images while using the standard microscopic set up available in most laboratories. It allows use of any fluorescent molecule (including quantum dots) that blinks or photobleaches to pick fluorophores out of the fluorescent background, and it can work with thousands of fluorophores in an image.

This image analysis algorithm can localize quantized drops in fluorescence even with a background to create super-resolution images from standard photobleaching movies. It does not require special photo-switchable fluorophores (PALM, STORM) or fluorophores with long-lived dark states (dSTORM). The data acquisition is accomplished in minutes instead of hours and it can use standard laboratory microscopic set up with any fluorescent molecule. The algorithm processes a movie in which fluorescent molecules or particles are photobleaching or blinking and looks for individual photobleaching events that can be located with high resolution (i.e. the position of the fluorescent molecules). A composite image with resolution much higher than the original movie can then be generated.

Noise is reduced by rejecting fluorophores that are poorly fit by a Gaussian and frames are averaged.

Benefits

The image analysis algorithm to create super-resolution images using standard photobleaching and blinking movies provides the following benefits:

• Uses standard laboratory equipment.
• Enables sub-cellular structure examination
• Can be used with any fluorescent molecule that can be detected by a CCD camera. It can use high labeling densities
• Fast data acquisition 

Dr. Kuhlman from the University of Illinois at Urbana-Champaign has created a new system that provides a single editing tool for a variety of genome modifications. This method has the advantage over CRISPR-Cas9 system in not requiring prior modification of the genome and requires only locus-specific primers for genome editing.

This technology is an optimized Landing Pad system that can use positive and negative selection to select for insertion, deletion, and knockout events at genomic targets using specific primers contain homology region A, B, and C. This approach provides a powerful tool for manipulating the genome.

Benefits

• The Landing-Pad system can integrate large fragments of DNA, can be targeted anywhere in the genome and ab be applicable to deletions and knockouts as well.
• Unlike the CRISPR-Cas9 system, this technique does not require a special promoter for gene expression.
• The Landing-Pad system can target different loci with only locus-specific primer design.

Researchers and practitioners in university, industrial and national labs increasingly rely on low temperatures in the development and study of products. For example, pharmaceutical companies routinely use low temperatures to study disease targets and markers.

The biomolecules are always in buffers. Since the pH of buffers changes dramatically upon lowering temperature, the biolmolecules are not the same pH when they are at the room temperature. The change of pH can dramatically change the structural and functional properties of the biomolecules so that information obtained at low temperature does not reflect its properties at room temperature or physiological temperature. Additionally, since the exact pH buffer at different temperatures is not often predictable, the results obtained using a temperature variable pH buffer cannot easily correlate back to the pH at room temperature.

This invention is a new design of pH buffers that show negligible pH change upon cooling to low temperatures, including cryotempatures. This invention solves the problem of significant change in apparent pH of a glycerol solution of common biological buffers upon cooling to cryotemperature. By combining a buffer that increases pH upon cooling with one that decreases pH upon cooling, the apparent pH change upon cooling to cryotemperature is minimized.