Image-Processing Software for High-Throughput Quantification of Colony Luminescence

ABSTRACT Many microbiological assays include colonies that produce a luminescent or fluorescent (here generalized as “luminescent”) signal, often in the form of luminescent halos around the colonies. These signals are used as reporters for a trait of interest; therefore, exact measurements of the lu...

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Autores principales:	Eyal Dafni, Iddo Weiner, Noam Shahar, Tamir Tuller, Iftach Yacoby
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2019
Materias:	fluorescent-image analysis microbial method software Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/299adca985f54093a01a00e794ad446c
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spelling	oai:doaj.org-article:299adca985f54093a01a00e794ad446c2021-11-15T15:22:04ZImage-Processing Software for High-Throughput Quantification of Colony Luminescence10.1128/mSphere.00676-182379-5042https://doaj.org/article/299adca985f54093a01a00e794ad446c2019-02-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00676-18https://doaj.org/toc/2379-5042ABSTRACT Many microbiological assays include colonies that produce a luminescent or fluorescent (here generalized as “luminescent”) signal, often in the form of luminescent halos around the colonies. These signals are used as reporters for a trait of interest; therefore, exact measurements of the luminescence are often desired. However, there is currently a lack of high-throughput methods for analyzing these assays, as common automatic image analysis tools are unsuitable for identifying these halos in the presence of the inherent biological noise. In this work, we have developed CFQuant—automatic, high-throughput software for the analysis of images from colony luminescence assays. CFQuant overcomes the problems of automatic identification by relying on the luminescence halo's expected shape and provides measurements of several features of the colonies and halos. We examined the performance of CFQuant using one such colony luminescence assay, where we achieved a high correlation (R = 0.85) between the measurements of CFQuant and known protein expression levels. This demonstrates CFQuant's potential as a fast and reliable tool for analysis of colony luminescence assays. IMPORTANCE Luminescent markers are widely used as reporters for various biologically interesting traits. In colony luminescence assays, the levels of luminescence around each colony can be used to compare the levels of traits of interest for different strains, treatments, etc., using quantitative measurements of the luminescence. However, automatic methods of obtaining this data are underdeveloped, making this a laborious manual process, especially in analyzing large numbers of colonies. The significance of this work is in developing an automatic, high-throughput tool for quantitative analysis of colony luminescence assays, which will allow fast collection of qualitative data from these assays and thus increase their overall usability.Eyal DafniIddo WeinerNoam ShaharTamir TullerIftach YacobyAmerican Society for Microbiologyarticlefluorescent-image analysismicrobial methodsoftwareMicrobiologyQR1-502ENmSphere, Vol 4, Iss 1 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	fluorescent-image analysis microbial method software Microbiology QR1-502
spellingShingle	fluorescent-image analysis microbial method software Microbiology QR1-502 Eyal Dafni Iddo Weiner Noam Shahar Tamir Tuller Iftach Yacoby Image-Processing Software for High-Throughput Quantification of Colony Luminescence
description	ABSTRACT Many microbiological assays include colonies that produce a luminescent or fluorescent (here generalized as “luminescent”) signal, often in the form of luminescent halos around the colonies. These signals are used as reporters for a trait of interest; therefore, exact measurements of the luminescence are often desired. However, there is currently a lack of high-throughput methods for analyzing these assays, as common automatic image analysis tools are unsuitable for identifying these halos in the presence of the inherent biological noise. In this work, we have developed CFQuant—automatic, high-throughput software for the analysis of images from colony luminescence assays. CFQuant overcomes the problems of automatic identification by relying on the luminescence halo's expected shape and provides measurements of several features of the colonies and halos. We examined the performance of CFQuant using one such colony luminescence assay, where we achieved a high correlation (R = 0.85) between the measurements of CFQuant and known protein expression levels. This demonstrates CFQuant's potential as a fast and reliable tool for analysis of colony luminescence assays. IMPORTANCE Luminescent markers are widely used as reporters for various biologically interesting traits. In colony luminescence assays, the levels of luminescence around each colony can be used to compare the levels of traits of interest for different strains, treatments, etc., using quantitative measurements of the luminescence. However, automatic methods of obtaining this data are underdeveloped, making this a laborious manual process, especially in analyzing large numbers of colonies. The significance of this work is in developing an automatic, high-throughput tool for quantitative analysis of colony luminescence assays, which will allow fast collection of qualitative data from these assays and thus increase their overall usability.
format	article
author	Eyal Dafni Iddo Weiner Noam Shahar Tamir Tuller Iftach Yacoby
author_facet	Eyal Dafni Iddo Weiner Noam Shahar Tamir Tuller Iftach Yacoby
author_sort	Eyal Dafni
title	Image-Processing Software for High-Throughput Quantification of Colony Luminescence
title_short	Image-Processing Software for High-Throughput Quantification of Colony Luminescence
title_full	Image-Processing Software for High-Throughput Quantification of Colony Luminescence
title_fullStr	Image-Processing Software for High-Throughput Quantification of Colony Luminescence
title_full_unstemmed	Image-Processing Software for High-Throughput Quantification of Colony Luminescence
title_sort	image-processing software for high-throughput quantification of colony luminescence
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/299adca985f54093a01a00e794ad446c
work_keys_str_mv	AT eyaldafni imageprocessingsoftwareforhighthroughputquantificationofcolonyluminescence AT iddoweiner imageprocessingsoftwareforhighthroughputquantificationofcolonyluminescence AT noamshahar imageprocessingsoftwareforhighthroughputquantificationofcolonyluminescence AT tamirtuller imageprocessingsoftwareforhighthroughputquantificationofcolonyluminescence AT iftachyacoby imageprocessingsoftwareforhighthroughputquantificationofcolonyluminescence
_version_	1718428059731755008

Image-Processing Software for High-Throughput Quantification of Colony Luminescence

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