NDLI: Expanding the available assays: adapting and validating In-Cell Westerns in microfluidic devices for cell-based assays.

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Expanding the available assays: adapting and validating In-Cell Westerns in microfluidic devices for cell-based assays.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Paguirigan, Amy L. Puccinelli, John P. Su, Xiaojing Beebe, David J.
Description	Country affiliation: United States Author Affiliation: Paguirigan AL ( Department of Biomedical Engineering, Wisconsin Institutes for Medical Research, University of Wisconsin-Madison, Madison, Wisconsin 53726, USA.)
Abstract	Microfluidic methods for cellular studies can significantly reduce costs due to reduced reagent and biological specimen requirements compared with many traditional culture techniques. However, current types of readouts are limited and this lack of suitable readouts for microfluidic cultures has significantly hindered the application of microfluidics for cell-based assays. The In-Cell Western (ICW) technique uses quantitative immunocytochemistry and a laser scanner to provide an in situ measure of protein quantities in cells grown in microfluidic channels of arbitrary geometries. The use of ICWs in microfluidic channels was validated by a detailed comparison with current macroscale methods and shown to have excellent correlation. Transforming growth factor-ß-induced epithelial-to-mesenchymal transition of an epithelial cell line was used as an example for further validation of the technique as a readout for soluble-factor-based assays performed in high-throughput microfluidic channels. The use of passive pumping for sample delivery and laser scanning for analysis opens the door to high-throughput quantitative microfluidic cell-based assays that integrate seamlessly with existing high-throughput infrastructure.
File Format	HTM / HTML
ISSN	1540658X
e-ISSN	15578127
DOI	10.1089/adt.2010.0274
Journal	ASSAY and Drug Development Technologies
Issue Number	5
Volume Number	8
Language	English
Publisher	Mary Ann Liebert, Inc.
Publisher Date	2010-10-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Discipline Analytical Discipline Pharmacology Discipline Chemistry Techniques High-throughput Screening Assays Immunoassay Microfluidic Analytical Techniques Antibodies Immunology Blotting, Western Cell Culture Techniques Cell Line Epithelial-mesenchymal Transition Image Processing, Computer-assisted Reproducibility Of Results Transforming Growth Factor Beta Pharmacology Research Support, N.i.h., Extramural Research Support, U.s. Gov't, Non-p.h.s. Validation Studies
Content Type	Text
Resource Type	Article
Subject	Drug Discovery Molecular Medicine

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Expanding the Available Assays: Adapting and Validating In-Cell Westerns in Microfluidic Devices for Cell-Based Assays

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Expanding the Available Assays: Adapting and Validating In-Cell Westerns in Microfluidic Devices for Cell-Based Assays

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