Evaluating single molecule detection methods for microarrays with high dynamic range for quantitative single cell analysis
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Published version
Author(s)
Salehi-Reyhani, S
Type
Journal Article
Abstract
Single molecule microarrays have been used in quantitative proteomics, in particular, single cell analysis requiring high sensitivity and ultra-low limits of detection. In this paper, several image analysis methods are evaluated for their ability to accurately enumerate single molecules bound to a microarray spot. Crucially, protein abundance in single cells can vary significantly and may span several orders of magnitude. This poses a challenge to single molecule image analysis. In order to quantitatively assess the performance of each method, synthetic image datasets are generated with known ground truth whereby the number of single molecules varies over 5 orders of magnitude with a range of signal to noise ratios. Experiments were performed on synthetic datasets whereby the number of single molecules per spot corresponds to realistic single cell distributions whose ground truth summary statistics are known. The methods of image analysis are assessed in their ability to accurately estimate the distribution parameters. It is shown that super-resolution image analysis methods can significantly improve counting accuracy and better cope with single molecule congestion. The results highlight the challenge posed by quantitative single cell analysis and the implications to performing such analyses using microarray based approaches are discussed.
Date Issued
2017-12-20
Date Acceptance
2017-12-07
Citation
Scientific Reports, 2017, 7
ISSN
2045-2322
Publisher
Nature Publishing Group
Journal / Book Title
Scientific Reports
Volume
7
Copyright Statement
This article is licensed under a Creative Commons Attribution 4.0 International
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Commons license, and indicate if changes were made. Te images or other third party material in this
article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the
material. If material is not included in the article’s Creative Commons license and your intended use is not permitted
by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the
copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
© The Author(s) 2017
License, which permits use, sharing, adaptation, distribution and reproduction in any medium or
format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative
Commons license, and indicate if changes were made. Te images or other third party material in this
article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the
material. If material is not included in the article’s Creative Commons license and your intended use is not permitted
by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the
copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
© The Author(s) 2017
License URL
Publication Status
Published
Article Number
17957