ShEEP Request for A Namocell PALA Single CellDispenser

James J Peters Va Medical Center, Bronx NY

Investigators

Abstract

We are requesting a Namocell PALA Single Cell Dispenser, a benchtop instrument that combines innovative microfluidics technology with flow cytometry and liquid dispensing. It can sort and dispense single cells into 96 or 384-well plates in minutes. Its low-pressure operation (<2 psi) preserves cell viability as well as integrity. The ability to isolate specific cell populations is invaluable for modern basic and applied biomedical research. Microfluidic droplet dispensers provide for miniaturized, high-throughput isolation of single cells in a microenvironment, which can be leveraged for ultra-sensitive and rapid assays. These units are ideal for a wide range of single cell applications including cell cloning with improved outgrowth, generation of high-yield single cell genomics libraries, rapid bulk sorting of specific target cells or rare cell populations, and recovery of single cells from as few as 100 cells. Single cell dispensers are currently in use in several academic and industry settings. Recent acquisition of Namocell Single Cell Dispensers by federally supported facilities include NIH/NIDA and Arkansas Little Rock VA. Data obtained with the PALA Single Cell Dispenser will have applications for both cell culture as well as rodent model studies of human disease. As such, we expect it will aid us in deciphering how these diseases develop or progress as well as help in identifying new treatment opportunities for Veterans with these diseases. The expected research applications for the Namocell PALA Single Cell Dispenser at JJP VAMC will range from bulk sorting of homogenous cell populations to single cell isolation. Bulk sorting is useful for initial interrogations of highly enriched, specific cell types. We expect JJP VAMC investigators who use rodent models of human diseases including those with neurological, renal, metabolic, or cancer relevance will use the PALA for bulk sorting of specific or rare cell populations, particularly those residing in organs with heterogenous cell types [Major Users: JG, CC, GP, RR]. Bulk sorting is also useful for identifying cells manipulated in culture that are marked by fluorescent activity, such as transfected cells expressing specific fluorescent proteins (FP). The dual laser system in the PALA will allow for selection of cells expressing dual exogenous proteins with distinct FP surrogates without the need for prolonged drug selection, which otherwise can introduce confounding factors [Major Users: JG, CC, RR]. The Namocell PALA Single Cell Dispenser will also be used for purification of single cells. Single-cell analysis can discriminate signal from noise in a heterogeneous cell population and thereby facilitate multiomic studies of rare cell populations obtained from tissues or blood of healthy or disease animal models [Major Users: JG, CC, GP, RR]. The PALA will also facilitate high-throughput isolation of single cells in a defined microenvironment, which can be leveraged for ultra-sensitive and rapid assays [Major Users: JG, GP, RR]. Finally, we anticipate great interest in the use of the PALA for cloning of precision genetically engineered transformed cells or induced pluripotent stem cells (iPSC), which may offer insights into structure-function studies as well as human disease contributions from nonsynonymous variants identified in human genetic studies, respectively [Major Users: JG, CC, GP, RR].