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Title: Identification of fluorescent compounds with non-specific binding property via high throughput live cell microscopy
Authors: Nath, S.
Spencer, V.A.
Han, J.
Chang, H.
Zhang, K.
Fontenay, G.V.
Anderson, C.
Hyman, J.M.
Nilsen-Hamilton, M.
Chang, Y.-T. 
Parvin, B.
Issue Date: 5-Jan-2012
Citation: Nath, S., Spencer, V.A., Han, J., Chang, H., Zhang, K., Fontenay, G.V., Anderson, C., Hyman, J.M., Nilsen-Hamilton, M., Chang, Y.-T., Parvin, B. (2012-01-05). Identification of fluorescent compounds with non-specific binding property via high throughput live cell microscopy. PLoS ONE 7 (1) : -. ScholarBank@NUS Repository.
Abstract: Introduction: Compounds exhibiting low non-specific intracellular binding or non-stickiness are concomitant with rapid clearing and in high demand for live-cell imaging assays because they allow for intracellular receptor localization with a high signal/noise ratio. The non-stickiness property is particularly important for imaging intracellular receptors due to the equilibria involved. Method: Three mammalian cell lines with diverse genetic backgrounds were used to screen a combinatorial fluorescence library via high throughput live cell microscopy for potential ligands with high in- and out-flux properties. The binding properties of ligands identified from the first screen were subsequently validated on plant root hair. A correlative analysis was then performed between each ligand and its corresponding physiochemical and structural properties. Results: The non-stickiness property of each ligand was quantified as a function of the temporal uptake and retention on a cell-by-cell basis. Our data shows that (i) mammalian systems can serve as a pre-screening tool for complex plant species that are not amenable to high-throughput imaging; (ii) retention and spatial localization of chemical compounds vary within and between each cell line; and (iii) the structural similarities of compounds can infer their non-specific binding properties. Conclusion: We have validated a protocol for identifying chemical compounds with non-specific binding properties that is testable across diverse species. Further analysis reveals an overlap between the non-stickiness property and the structural similarity of compounds. The net result is a more robust screening assay for identifying desirable ligands that can be used to monitor intracellular localization. Several new applications of the screening protocol and results are also presented. © 2012 Nath et al.
Source Title: PLoS ONE
ISSN: 19326203
DOI: 10.1371/journal.pone.0028802
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