Identification of fluorescent compounds with non-specific binding property via high throughput live cell microscopy
dc.creator | Nath, S | |
dc.creator | Spencer, VA | |
dc.creator | Han, J | |
dc.creator | Chang, H | |
dc.creator | Zhang, K | |
dc.creator | Fontenay, GV | |
dc.creator | Anderson, C | |
dc.creator | Hyman, JM | |
dc.creator | Nilsen-Hamilton, M | |
dc.creator | Chang, YT | |
dc.creator | Parvin, B | |
dc.date.accessioned | 2021-01-31T22:39:43Z | |
dc.date.available | 2021-01-31T22:39:43Z | |
dc.date.issued | 2012-01-05 | |
dc.identifier.issn | 1932-6203 | |
dc.identifier.issn | 1932-6203 | |
dc.identifier.doi | http://dx.doi.org/10.34944/dspace/5457 | |
dc.identifier.other | 22242152 (pubmed) | |
dc.identifier.uri | http://hdl.handle.net/20.500.12613/5475 | |
dc.description.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. | |
dc.format.extent | e28802-e28802 | |
dc.language.iso | en | |
dc.relation.haspart | PLoS ONE | |
dc.relation.isreferencedby | Public Library of Science (PLoS) | |
dc.rights | CC BY | |
dc.subject | Animals | |
dc.subject | Arabidopsis | |
dc.subject | Cell Line | |
dc.subject | Cell Survival | |
dc.subject | Combinatorial Chemistry Techniques | |
dc.subject | Fluorescent Dyes | |
dc.subject | Humans | |
dc.subject | Ligands | |
dc.subject | Mice | |
dc.subject | Microscopy | |
dc.subject | Small Molecule Libraries | |
dc.title | Identification of fluorescent compounds with non-specific binding property via high throughput live cell microscopy | |
dc.type | Article | |
dc.type.genre | Journal Article | |
dc.relation.doi | 10.1371/journal.pone.0028802 | |
dc.ada.note | For Americans with Disabilities Act (ADA) accommodation, including help with reading this content, please contact scholarshare@temple.edu | |
dc.date.updated | 2021-01-31T22:39:39Z | |
refterms.dateFOA | 2021-01-31T22:39:44Z |