A multiwell plate format microfluidic immobilization chip for high-content imaging of whole animals

Advancements in precise genetic manipulation have helped biologists to identify new drug targets and in vivo disease mechanisms using small animal modelssuch as CelegansHuman diseases pathophysiologies are reproduced in Celegans expressing human disease genes inside the animalGreat opportunities are provided by the recent surge is genetic tools for animal modelsbut the lack of high content screeningHCStechnologies precluded these models from screening for subtle phenotypes that better recapitulate the human disease situationsDevelopment of novel technologies will enable the use of such modelsat the same cost and speed of in vitro assaysto discover new drug targets and understand mode of action for new compounds in vivoDrBen Yakar Lab at The University of Texas at Austin has developed a novel large scale microfluidic chip that can imageanimals frompopulations using a proprietary channel designAn efficient image acquisition and analysis algorithms can screen a whole chip withinmina record speed that isfaster than manual imagingTo translate this lab prototype into marketplacethis SBIR Phase I application proposes to develop a beta model vivoChipx that will be with SBS formatcompatible with automationlighter weightless expensiveand user friendly to operate with the new top gasket designThe new chip design will be bonded to a thin glass substrate at the bottom to enable improved imaging using high resolution objectivesThe proposed microfluidic chip will incorporate a machined top plastic with custom designed wells in a micro titer format for easy integration to liquid handling systems for the high throughput screenHTSand avoid substrate bendingand chip handling errorsIn Aimwe plan to develop a beta design of the vivoChipx device with the topplastic piecefabricate a thin PDMS layer using soft lithographyand bond the interfaces using appropriate UVcured glue and plasma treatmentThe beta model will be operated with a new top gasket with an improved sealing mechanismCelegans populations will be trapped inside the channels to characterize the variability in the trapping efficiency using four chips and following the standard operational proceduresSOPsIn Aimwe will develop an automated acquisition algorithm with BioTek to image Celegans with high reslution objectives having a sub cellular expression of fluorescent proteins and achieve an assay quality ZAchieving these milestones in Phase Iwe will be able to reduce the current cost of the chip byfolds and standardize the vivoChipx for all commercially available HCS instrumentsIn Phase IIwe will develop a fully automated vivoLoader to replace our current semi automated worm handling procedures of liquid handling and an automated image analysis platformvivoAnalyzerthat will identify subtle fluorescent phenotype in low expressing CelegansUsing our ongoing research collaborationwe plan to apply our screening technology to develop neurotoxicity and neurodegeneration assays to be able to screen novel compounds from large pharmaceutical companiesSupport from industry partners will help us to translate the prototype into a product Project NarrativeThe proposed paradigm shifting technology will allow the use of small animal models to discover new chemical compounds with improved in vivo toxicity profiles and efficacy for preventing or delaying age dependent declines in their healthImplementing whole organism screens at an early stage of drug discovery can lead to identifying novel drug targets and disease mechanisms in vivo that will have high translation probability during drug developmentnecessary for chronic illnesses that suffer from the slow and poor discovery process