SeeTrue: The Microinjection Colored Needle For Enhanced Visibility and Anti-Clogging Capabilities

Project Summary Microinjection is a widely used transgenic engineering technique that underpins genetic manipulationexperimentationand knowledge discovery across virtually all areas of biomedical scienceIt consists of using an industry standardtransparentclear glass microneedleISNto manually inject foreign materialse gRNA and DNAinto biological cellsThis thirty year old technology has driven advances in the fundamental research of stem cell gene manipulationcreated the intra cytoplasmic sperm injection method as part of invitro fertilization cycle therapyand allowed extensive human disease prevention modeling via pathophysiological investigationsThree design induced constraintsoften attributed to poor user techniqueaffect the technique s efficacy and experimental rigor and reproducibilitythe transparent needle tip creates low contrast visibility in vitroleaving the user unable to view and track the penetrating needle tipcytoplasmic materiale gembryo yolkcan clog the needle tipdiminishing volume injected over timeandmanual creation of each needle via industry standard pipette pullers introduces calibration variability with every new needle usedAlthough other foreign material introduction methods existsuch as electroporationchemical membrane permeabilizationand automatedrobotic systemsnone have proven responsive to addressing these key design constraintsThe objective of this proposalprototyping a colored tipmicrocapillary needle with anti clogging properties as a minimal viable productMVPbridges an important need in the biomedical industryThe working hypothesis is that the patentedSeeTrue MVP will simply and powerfully increase the accuracy of the microinjection technique as compared to ISNs by aimproving the efficacy of delivering material to the embryoand breducing variability in the volume injectedThe MVP will be testedand its efficacy compared to ISN s in the Zebrafish embryo biological system by both experiencedmonthsand inexperiencedless than one monthZebrafish embryo microinjectorsThe MVP fabrication and testing will be achieved in the following two aimsAimFabricate a nanoparticle coatedcolored glass MVP with anti clogging capabilitiesAimTest the colored and anti clogging SeeTrue MPV using Zebrafish embryosThe MVP s ability to increase an injectorsaccuracy will be quantified by measuring the amount of material delivered to the embryo and by the degree of variability in the amount of material delivered to embryos following multiple microinjectionsIn additiona comparative user satisfaction survey will be administered to all testersThe proposed SeeTrue MVP introduces the simpleelegantdesign innovations of enhanced visibility and anti clogging properties to an establishedfundamental techniqueIt has the potential to correct known rigor and reproducibilitytechniquedeficits and increase the efficacy of microinjection biomedical transgenic researchThis willin turn positively impact foundational human health related research and medical applications Project Narrative The proposed researcha prototype for a coloredanticlogging microinjection needleis relevant to public health research because it represents a new and substantive departure from the status quoThis prototype has the potential to increase the rigorreproducibilityand efficacy of a broad range of research that uses the manual microinjection technique by introducing two design improvements that will improve the accuracy of the techniqueThis is relevant to the part of NIH s mission that pertains to both basic human health transgenic research and medical applications