ShEEP Request for A Namocell PALA Single CellDispenser

我们正在申请一台Namocell PALA单细胞分配器，这是一台台式仪器， 将创新的微流体技术与流式细胞术和液体分配相结合。它可以 在几分钟内将单细胞分选并分配到96或384孔板中。其低压运行（<2 psi）保持细胞活力以及完整性。分离特定细胞群的能力是 对现代基础和应用生物医学研究来说是无价的。微流体液滴分配器 在微环境中提供单细胞的小型化、高通量分离， 可以用于超灵敏和快速的测定。这些单位是理想的广泛的 单细胞应用，包括具有改善的生长的细胞克隆，高产率的产生， 单细胞基因组文库，特定靶细胞或稀有细胞群的快速批量分选， 以及从少至100个细胞中回收单个细胞。 单细胞分配器目前在几个学术和工业环境中使用。最近 由联邦政府支持的机构采购的Namocell单细胞分配器包括 NIH/NIDA和阿肯色州弗吉尼亚州小石城。使用PALA单细胞分配器获得的数据将 可用于细胞培养以及人类疾病的啮齿动物模型研究。因此，在本发明中， 我们希望它能帮助我们破译这些疾病是如何发展或进展的， 为患有这些疾病的退伍军人寻找新的治疗机会。 Namocell PALA单细胞分配器在JJP VAMC的预期研究应用 将从同质细胞群的批量分选到单细胞分离。批量分拣是 可用于高度富集的特定细胞类型的初始询问。我们希望JJP VAMC 研究人员使用人类疾病的啮齿动物模型，包括那些神经，肾脏， 代谢或癌症相关性将使用PALA进行特定或稀有细胞的批量分选 群体，特别是存在于具有异质细胞类型的器官中的那些[主要用户：JG， CC、GP、RR]。批量分选也可用于鉴定在培养物中操作的细胞， 通过荧光活性，如表达特异性荧光蛋白（FP）的转染细胞。 PALA中的双激光系统将允许选择表达双外源基因的细胞。 具有不同FP替代物的蛋白质，而不需要延长的药物选择， 否则可能引入混杂因素[主要用户：JG、CC、RR]。 Namocell PALA单细胞分配器也将用于纯化单细胞。 单细胞分析可以区分异质细胞群体中的信号与噪声， 从而促进从人的组织或血液中获得的稀有细胞群的多组学研究。 健康或疾病动物模型[主要用户：JG、CC、GP、RR]。PALA还将促进 在定义的微环境中高通量分离单细胞，可利用 用于超灵敏和快速测定[主要用户：JG、GP、RR]。最后，我们期待伟大的 对使用PALA克隆精确遗传工程转化细胞的兴趣 或诱导多能干细胞（iPSC），这可能为结构-功能研究提供见解 以及来自人类遗传学中鉴定的非同义变体的人类疾病贡献 研究[主要用户：JG、CC、GP、RR]。

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].