| Literature DB >> 31668230 |
David H Abraham1, Matthew M Anttila1, Luke A Gallion1, Brae V Petersen1, Angela Proctor1, Nancy L Allbritton2.
Abstract
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 10-20mol.Entities:
Keywords: Capillary electrophoresis; Cell array; Chemical cytometry; Enzyme activity measurement; Fluorescence detection; Laser-based lysis; Single cell
Mesh:
Year: 2019 PMID: 31668230 PMCID: PMC6933539 DOI: 10.1016/bs.mie.2019.06.016
Source DB: PubMed Journal: Methods Enzymol ISSN: 0076-6879 Impact factor: 1.600