Applications and Caveats on the Utilization of DNA-Specific Probes in Cell-Based Assays.

Applications and Caveats on the Utilization of DNA-Specific Probes in Cell-Based Assays.
Methods Mol Biol. 2018;1683:3-19
Authors: Edward R
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.
PMID: 29082483 [PubMed - in process]

Source: Mol Biol Cell - October 31, 2017 Category: Molecular Biology Authors: Edward R Tags: Methods Mol Biol Source Type: research