Radiofrequency-multiplexed parallel flow cytometer for high content screening and drug discovery

DESCRIPTION provided by applicant Drug discovery is an extremely lengthy and expensive process On average drugs today cost more than $ billion to develop and take more than a decade to reach the market Most drugs today are discovered using high throughput screening in which hundreds of thousands to millions of trial compounds are assayed against cells to determine if they interact with a disease target This is typically performed using at the well level using techniques such as fluorescence chemiluminescence and optical absorbance to measure a cell populationandapos s average response to a drug While these techniques are high throughput they lack the ability to interrogate wells at the single cell level which provides a much more comprehensive picture of the phenotypic drug cell interaction than population averaged measurement do Flow cytometry is a well established and widely used technique used in most areas of cell biology that provides multi parameter cell level phenotypic information by measuring optical scatter and fluorescence information from individual cells in flow at a high throughput cells second While this cellular throughput is high the sample throughput of flow cytometers is low compared to fluorescence plate readers for example This speed limitation is primarily due to the serial sample handling approach employed by flow cytometers If this speed limitation could be eliminated flow cytometry could be used in primary screening assays The high content phenotypic information obtained by using flow cytometry early in the drug discovery process would enable researchers to more readily understand a candidate compoundandapos s complex intra and inter cellular effects This transformation in the compound screening process would increase the efficiency of drug discovery by elucidating complex drug cell interactions at an earlier stage ultimately reducing the overall cost and time to market of new drugs Here we propose to develop a parallel flow cytometer system using a novel optical technology known as Fluorescence Imaging using Radiofrequency tagged Emission or FIRE FIRE is a high speed optical technique that enables a single photomultiplier tube detector to measure fluorescence or optical scatter signals from multiple points on a sample using radiofrequency domain multiplexing We will use a modified FIRE optical system to probe fluorescence and scatter from cells flowing in parallel flow channels This instrument will simultaneously collect flow cytometry data from samples aspirated from well plates using a multi probe autosampler We will design construct and characterize the system using standard protocols and perform a relevant screening assay to test its performance This system will enable a well plate to be sampled in less than minutes ultimately enabling high content compound screening of more than wells per day using flow cytometry PUBLIC HEALTH RELEVANCE An instrument capable of analyzing cells flowing in parallel is proposed for improving the drug discovery process This instrument will enable candidate drug compounds to be screened at a rate approximately times faster than the current state of the art which will ultimately improve the efficiency at which researchers discover new pharmaceuticals