Chapter title |
Applications and Caveats on the Utilization of DNA-Specific Probes in Cell-Based Assays
|
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Chapter number | 1 |
Book title |
High Content Screening
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-7357-6_1 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7355-2, 978-1-4939-7357-6
|
Authors |
Roy Edward |
Abstract |
To perform cell-based assays using fluorescence as the readout there is a fundamental need to identify individual cellular objects. In the majority of cases this requires the addition of a DNA dye or so-called nuclear counterstain and these have become integral to assay design. End-point assays can use live or fixed cells and thus it is beneficial if such reagents are cell membrane-permeant.Further, membrane-permeant DNA dyes can open new opportunities in dynamic real time assays with caveats according to the impact of their interaction with the chromatin in live cells. As cell-based assays offer information on the in vitro toxicity of treatments, cell viability has become a basic readout and cell membrane-impermeant fluorescent DNA-specific dyes can provide this information.In the case of both nuclear counterstaining and viability reporting, it is beneficial if the DNA dyes employed are suitably spectrally separated to permit multi-color experimental design. Methods will be described for these two important assay readouts. |
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