Human cell chips: adapting DNA microarray spotting technology to cell-based imaging assays.

Here we describe human spotted cell chips, a technology for determining cellular state across arrays of cells subjected to chemical or genetic perturbation. Cells are grown and treated under standard tissue culture conditions before being fixed and printed onto replicate glass slides, effectively de...

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Autores principales:	Traver Hart, Alice Zhao, Ankit Garg, Swetha Bolusani, Edward M Marcotte
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2009
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/2c1acf4489264f33a18b608658bacb02
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spelling	oai:doaj.org-article:2c1acf4489264f33a18b608658bacb022021-11-25T06:28:28ZHuman cell chips: adapting DNA microarray spotting technology to cell-based imaging assays.1932-620310.1371/journal.pone.0007088https://doaj.org/article/2c1acf4489264f33a18b608658bacb022009-10-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/19862318/?tool=EBIhttps://doaj.org/toc/1932-6203Here we describe human spotted cell chips, a technology for determining cellular state across arrays of cells subjected to chemical or genetic perturbation. Cells are grown and treated under standard tissue culture conditions before being fixed and printed onto replicate glass slides, effectively decoupling the experimental conditions from the assay technique. Each slide is then probed using immunofluorescence or other optical reporter and assayed by automated microscopy. We show potential applications of the cell chip by assaying HeLa and A549 samples for changes in target protein abundance (of the dsRNA-activated protein kinase PKR), subcellular localization (nuclear translocation of NFkappaB) and activation state (phosphorylation of STAT1 and of the p38 and JNK stress kinases) in response to treatment by several chemical effectors (anisomycin, TNFalpha, and interferon), and we demonstrate scalability by printing a chip with approximately 4,700 discrete samples of HeLa cells. Coupling this technology to high-throughput methods for culturing and treating cell lines could enable researchers to examine the impact of exogenous effectors on the same population of experimentally treated cells across multiple reporter targets potentially representing a variety of molecular systems, thus producing a highly multiplexed dataset with minimized experimental variance and at reduced reagent cost compared to alternative techniques. The ability to prepare and store chips also allows researchers to follow up on observations gleaned from initial screens with maximal repeatability.Traver HartAlice ZhaoAnkit GargSwetha BolusaniEdward M MarcottePublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 4, Iss 10, p e7088 (2009)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Traver Hart Alice Zhao Ankit Garg Swetha Bolusani Edward M Marcotte Human cell chips: adapting DNA microarray spotting technology to cell-based imaging assays.
description	Here we describe human spotted cell chips, a technology for determining cellular state across arrays of cells subjected to chemical or genetic perturbation. Cells are grown and treated under standard tissue culture conditions before being fixed and printed onto replicate glass slides, effectively decoupling the experimental conditions from the assay technique. Each slide is then probed using immunofluorescence or other optical reporter and assayed by automated microscopy. We show potential applications of the cell chip by assaying HeLa and A549 samples for changes in target protein abundance (of the dsRNA-activated protein kinase PKR), subcellular localization (nuclear translocation of NFkappaB) and activation state (phosphorylation of STAT1 and of the p38 and JNK stress kinases) in response to treatment by several chemical effectors (anisomycin, TNFalpha, and interferon), and we demonstrate scalability by printing a chip with approximately 4,700 discrete samples of HeLa cells. Coupling this technology to high-throughput methods for culturing and treating cell lines could enable researchers to examine the impact of exogenous effectors on the same population of experimentally treated cells across multiple reporter targets potentially representing a variety of molecular systems, thus producing a highly multiplexed dataset with minimized experimental variance and at reduced reagent cost compared to alternative techniques. The ability to prepare and store chips also allows researchers to follow up on observations gleaned from initial screens with maximal repeatability.
format	article
author	Traver Hart Alice Zhao Ankit Garg Swetha Bolusani Edward M Marcotte
author_facet	Traver Hart Alice Zhao Ankit Garg Swetha Bolusani Edward M Marcotte
author_sort	Traver Hart
title	Human cell chips: adapting DNA microarray spotting technology to cell-based imaging assays.
title_short	Human cell chips: adapting DNA microarray spotting technology to cell-based imaging assays.
title_full	Human cell chips: adapting DNA microarray spotting technology to cell-based imaging assays.
title_fullStr	Human cell chips: adapting DNA microarray spotting technology to cell-based imaging assays.
title_full_unstemmed	Human cell chips: adapting DNA microarray spotting technology to cell-based imaging assays.
title_sort	human cell chips: adapting dna microarray spotting technology to cell-based imaging assays.
publisher	Public Library of Science (PLoS)
publishDate	2009
url	https://doaj.org/article/2c1acf4489264f33a18b608658bacb02
work_keys_str_mv	AT traverhart humancellchipsadaptingdnamicroarrayspottingtechnologytocellbasedimagingassays AT alicezhao humancellchipsadaptingdnamicroarrayspottingtechnologytocellbasedimagingassays AT ankitgarg humancellchipsadaptingdnamicroarrayspottingtechnologytocellbasedimagingassays AT swethabolusani humancellchipsadaptingdnamicroarrayspottingtechnologytocellbasedimagingassays AT edwardmmarcotte humancellchipsadaptingdnamicroarrayspottingtechnologytocellbasedimagingassays
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Human cell chips: adapting DNA microarray spotting technology to cell-based imaging assays.

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