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Analysis of image-based phenotypic parameters for high throughput gene perturbation assays

Paper ID Volume ID Publish Year Pages File Format Full-Text
15019 1367 2015 7 PDF Available
Title
Analysis of image-based phenotypic parameters for high throughput gene perturbation assays
Abstract

•Image-based multiplexing assay was attempted for siRNA library screening.•Diverse orthogonal phenotypic parameters were retrieved from a single screening.•Image-based cell count provided better resolution than the viability measure.•Geneset analysis revealed biological implications of various image-based parameters.

Although image-based phenotypic assays are considered a powerful tool for siRNA library screening, the reproducibility and biological implications of various image-based assays are not well-characterized in a systematic manner. Here, we compared the resolution of high throughput assays of image-based cell count and typical cell viability measures for cancer samples. It was found that the optimal plating density of cells was important to obtain maximal resolution in both types of assays. In general, cell counting provided better resolution than the cell viability measure in diverse batches of siRNAs. In addition to cell count, diverse image-based measures were simultaneously collected from a single screening and showed good reproducibility in repetitions. They were classified into a few functional categories according to biological process, based on the differential patterns of hit (i.e., siRNAs) prioritization from the same screening data. The presented systematic analyses of image-based parameters provide new insight to a multitude of applications and better biological interpretation of high content cell-based assays.

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Keywords
siRNA screening; Gene perturbation; Image-based assay; Phenotypic parameter
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Analysis of image-based phenotypic parameters for high throughput gene perturbation assays
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Publisher
Database: Elsevier - ScienceDirect
Journal: Computational Biology and Chemistry - Volume 58, October 2015, Pages 192–198
Authors
, , , , , ,
Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us