Detection quantitation and analysis of extracellular vessicles using reflective phase grating technology

Over the past or years it has become clear that extracellular vesicles EVs play a far more important role in patient physiology than previously thought There are now numerous examples of studies showing that EVs have a role in cell cell communication in neuronal cells may be involved in drug transport across the blood brain barrier and impact a variety of signaling mechanism because it is known that EVs can cross the blood brain barrier The fundamental problem which is the most common thread in the entire literature is that EVs are hard to isolate difficult to identify very difficult to quantitate and providing classification information is very time consuming and costly EVs are defined as being from nm far below the diffraction limit of optical systems and therefore too small to image without the aid of electron microscopy This proposal presents a technology that is unique in that it can create an electronic signature of EVs and translate that into a pseudo image with precise details in the spatial domain It can quantitate particles and provide a distribution analysis but this distribution is based on single particle measurement not bulk measurements like dynamic light scatting The technology is based on the principle of reflective phase grating RFG a technology never before applied to biological systems but regardless is a mature and low cost technology that can be mass produced as it has for the Blu ray technology There are four key features of RFG technology particle resolution approaches nm analysis takes minutes not hours particles can be measured in liquid phase and material cost per sample approaches $ so the potential impact on research and clinical diagnostics is enormous We propose to add a fluorescence detector within the confines of this application which will provide origin classifier information critical to understanding the impact EVs have on physiology Exosomes are small components of cells that have unique properties and play an important role in the regulation of many drugs and treatments We now believe that these tiny particles may carry important information in drug addiction diseases that impact the brain such as Alzheimer s disease and other effects that might impact brain function Because these small components are far smaller than most technologies can detect them this project has developed a low cost technology that can rapidly identify and analyze these tiny components This new technology will provide a rapid method for researchers and clinicians to evaluation the impact of drugs on patients within a few minutes