Fluorescent nanofoldamers for multiplex flow cytometry

PROJECT SUMMARY ABSTRACT Polychromatic flow cytometryFCis one of the most powerful analytical techniques used in immunologybasic researchand clinical medicineIn basic researchFC is a primary tool for understanding disease development at the cellular and subcellular levelsIn the rapidly developing field of immunotherapyFC is an indispensable tool for monitoring the effectiveness of new therapies and identifying key cellular subpopulations using multiple biomarkers within a panelIt plays a critical role in several growing clinical applicationsdevelopment of vaccines for infectious diseasesincluding emerging diseases such as Ebola and Zikacharacterization of immunological cells and responses related to transplantation and treatment of graft versus host diseaseand development of vaccines and immune therapies to prevent and treat HIV infectionsIncreasing the number of spectrally distinct fluorophores for analysis of a single sample will give researchers greater flexibility and give clinicians a higher level of accuracy in the diagnosis and management of conditions where the size of the sample use for diagnosis is limited and for specimens with low cell numberssuch as those derived from fine needle aspirationlaparoscopycore biopsyand cerebrospinal fluidDue to these small sample sizesthere are often insufficient cell numbers for multiple panelsImprovements in optics and lasers have driven the recent introduction of new fluorophores for FCHowevermost of these new fluorophores have broad and often overlapping emission profilesmuch like traditional fluorophoresOverlapping spectra can be resolved by the combined use of bandpass filters and mathematical compensationoverlap subtractionhowever such compensation increases experimental errorreduces sensitivityand limits multiplexingThuseven with a prototypecolor instrumentleading investigators have only been able to implement aparameter paneladvances in reader technology have outstripped available capabilities in fluorophoresNIRvana Sciences is currently developing two families of tetrapyrrole fluorophores licensed from NC State UniversityThese fluorophores offer properties that make them excellent candidates for use in polychromatic FC including ultra violet violet excitationlong stokes shifts into the red and near infrared spectrums along with very narrow emissionsNIRvana Sciences and the Lindsey lab are developing a polymer platform that retains the intrinsic brightness of its dyes and has strong utility to the flow cytometry reagent market PROJECT NARRATIVEPUBLIC HEALTH RELEVANCE Flow cytometry is used routinely for both basic research and clinical diagnostic applicationsMany types of infectious diseases and autoimmunity are routinely diagnosed using flow cytometryand it is widely used in the development of vaccines and immunotherapies for cancerThis STTR project will advance the development of a new technology for creating bright and sharp dyes in the red to far red spectral regionwhich will greatly expand the suite ofcolorsavailable for multiplexed flow cytometry and allow for higher sensitivitysmaller sample sizeand higher throughput testing