An integrated instrument system for Patch-sequencing

State University New York Stony Brook, Stony Brook NY

Investigators

Abstract

Title: Integrated Patch-seq Instrument PROJECT SUMMARY This proposal requests funds for the purchase of an integrated Patch-seq instrument. The instrument will be used to perform multimodal analysis of the morphology, electrophysiology, and transcriptome of single cells. The instrument consists of a microscope (equipped with two-photon, epifluorescence, and differential interference optics), micromanipulators and electronics for the recording of the electrophysiological properties of single cells. Integration of these components is routine and does not affect the warranty of any component. The instrument is controlled via a single computer that will use a combination of propriety and open-source software for real-time recording and analysis of the cell’s electrophysiological and morphological properties. Currently, there is no instrument at Stony Brook University, or any nearby institution optimized to perform Patch-seq analysis. This instrument will enable researchers to perform Patch-seq analysis on a broad variety of tissues and cells. It is of particular use to neurobiologists and developmental biologists studying the nervous system but has application to a broad range of physiological and pre-clinical research programs. This instrument will allow the researchers to integrate familiar single cell analysis techniques (morphology and electrophysiology) with datasets created using high throughput single cell sequencing technology. The Patch-seq instrument has also the capability to support very innovative projects incorporating aspects of the network function of single neurons cell with high throughput single cell sequencing. The Institution is extremely supportive, providing $126K in resources to establish and support this instrument at Stony Brook University. Management and operation of the instrument will be provided by a faculty specialist, who will also assist users by either training them with the use of the instrument or by performing experiments in collaboration with participating laboratories. The instrument will be supported by the university’s Single Cell Genomics facility which will also perform cDNA library preparation and oversee sequencing. Bioinformatics specialists associated with the facility will assist with data analysis, where necessary.