Single cell transcriptional gene expression profiling is still highly plagued by limited starting material from single cells, time- consuming sample preparation, and lacks of game-changing platform that enables cells-in-answer-out analysis of an individual cell. To revolutionize this scenario, herein, we develop a very large scale integrated microfluidic single cell analyzer that allows rapid gene expression measurements within one hour over hundreds of single cells plus 24 genes via multiplex qPCR assay on a single chip in per run. In order to execute single cell gene analysis on chip without contaminations and loss of limited samples, we integrate functional microfluidic circuits for single cell isolation, lysis, reverse transcription, and multiple gene qPCR assays. We validate the capability of our single cell microprocessor of whole pathway analysis within a single cell by applying our platform to dissect single cell apoptosis related genes across over 4,000 malignant human breast tumor cells (MDA-MB-231) and 1,000 non-malignant breast tumor cells (MCF-7). By one single run, we can directly obtain the gene expression heat maps, over 24 key genes within 128 single cells, which cannot be obtained by any other existing techniques. Reliable and rapid systematic gene analysis by single cell microprocessor will transform current quantitative life sciences and precision medicine, which enable predictive and preventive medicine.
Project end date: 02/01/16