The broader impact/commercial potential of this Small Business Innovation Research (SBIR) project is to improve the efficiency of discovering new drugs by introducing a new type of instrument for fluorescent imaging of cells at high throughput. This innovation will allow pharmaceutical researchers to image individual cells at speeds more than an order of magnitude faster than the state-of-the-art instrumentation, which will improve the rate at which scientists run drug screening experiments. By taking high-resolution images of individual cells during these screening experiments, far more detailed information can be gleaned than using conventional high throughput methods, leading to a deeper scientific understanding of drug-cell interactions. This instrument will address the growing $15 billion drug discovery instrumentation market by providing a new type of high throughput readout capability for the drug discovery industry. Further, the ability to perform multiplexed multi-color fluorescent imaging of cells will reduce the number of experiments required to identify new pharmaceutical compounds, thereby improving the efficiency and reducing the cost of drug discovery, ultimately reducing the time-to-market of new drugs.<br/><br/>This SBIR Phase I project proposes to develop an imaging flow cytometer instrument to improve the speed of single-cell analysis in the field of drug discovery. While multi-parameter single-cell phenotypic analysis provides great information about the cellular interactions of a pharmaceutical compound, this type of experimental readout is too slow to perform on the large numbers of samples common in a drug discovery laboratory. By performing parallel fluorescent image analysis of cells in flow, this process can be accelerated such that more than 100,000 compounds can be screened each day in an automated laboratory setting. This project aims to develop an instrument capable of such high throughput image analysis using fluorescence, combining techniques and technologies adapted from the fields of flow cytometry and high-speed fluorescence microscopy to accomplish this throughput goal. The successful outcome of this project will yield an instrument capable of imaging cells flowing at meter per second flow rates with diffraction-limited spatial resolution using multiple fluorescent colors.