Design of an automated capillary electrophoresis platform for single-cell analysis

被引:6
作者
Abraham, David H. [1 ]
Anttila, Matthew M. [1 ]
Gallion, Luke A. [1 ]
Petersen, Brae V. [1 ]
Proctor, Angela [1 ]
Allbritton, Nancy L. [1 ,2 ]
机构
[1] Univ N Carolina, Dept Chem, Chapel Hill, NC 27515 USA
[2] Joint Dept Biomed Engn Univ North Carolina Chapel, Raleigh, NC 27599 USA
来源
ENZYME ACTIVITY IN SINGLE CELLS | 2019年 / 628卷
基金
美国国家卫生研究院; 美国国家科学基金会;
关键词
SPHINGOSINE KINASE-ACTIVITY; CHEMICAL CYTOMETRY; AKT ACTIVITY; SEPARATION; LYSIS; FLUORESCENCE; EFFICIENCY; SYSTEM;
D O I
10.1016/bs.mie.2019.06.016
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Single-cell analysis of cellular contents by highly sensitive analytical instruments is known as chemical cytometry. A chemical cytometer typically samples one cell at a time, quantifies the cellular contents of interest, and then processes and reports that data. Automation adds the potential to perform this entire sequence of events with minimal intervention, increasing throughput and repeatability. In this chapter, we discuss the design considerations for an automated capillary electrophoresis-based instrument for assay of enzymatic activity within single cells. We describe the key requirements of the microscope base and capillary electrophoresis platforms. We also provide detailed protocols and schematic designs of our cell isolation, lysis, sampling, and detection strategies. Additionally, we describe our signal processing and instrument automation workflows. The described automated system has demonstrated single-cell throughput at rates above 100cells/h and analyte limits of detection as low as 10(-20) mol.
引用
收藏
页码:191 / 221
页数:31
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