Cargando…

Digital, Rapid, Accurate, and Label-Free Enumeration of Viable Microorganisms Enabled by Custom-Built On-Glass-Slide Culturing Device and Microscopic Scanning

Accurately measuring the number of viable microorganisms plays an essential role in microbiological studies. Since the conventional agar method of enumerating visible colonies is time-consuming and not accurate, efforts have been made towards overcoming these limitations by counting the invisible mi...

Descripción completa

Detalles Bibliográficos
Autores principales: Song, Donghui, Liu, Haomin, Dong, Qiuchen, Bian, Zichao, Wu, Huixiang, Lei, Yu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6263752/
https://www.ncbi.nlm.nih.gov/pubmed/30384414
http://dx.doi.org/10.3390/s18113700
_version_ 1783375354679787520
author Song, Donghui
Liu, Haomin
Dong, Qiuchen
Bian, Zichao
Wu, Huixiang
Lei, Yu
author_facet Song, Donghui
Liu, Haomin
Dong, Qiuchen
Bian, Zichao
Wu, Huixiang
Lei, Yu
author_sort Song, Donghui
collection PubMed
description Accurately measuring the number of viable microorganisms plays an essential role in microbiological studies. Since the conventional agar method of enumerating visible colonies is time-consuming and not accurate, efforts have been made towards overcoming these limitations by counting the invisible micro-colonies. However, none of studies on micro-colony counting was able to save significant time or provide accurate results. Herein, we developed an on-glass-slide cell culture device that enables rapid formation of micro-colonies on a 0.38 mm-thick gel film without suffering from nutrient and oxygen deprivation during bacteria culturing. Employing a phase contrast imaging setup, we achieved rapid microscopic scanning of micro-colonies within a large sample area on the thin film without the need of fluorescent staining. Using Escherichia coli (E. coli) as a demonstration, our technique was able to shorten the culturing time to within 5 h and automatically enumerate the micro-colonies from the phase contrast images. Moreover, this method delivered more accurate counts than the conventional visible colony counting methods. Due to these advantages, this imaging-based micro-colony enumeration technique provides a new platform for the quantification of viable microorganisms.
format Online
Article
Text
id pubmed-6263752
institution National Center for Biotechnology Information
language English
publishDate 2018
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-62637522018-12-12 Digital, Rapid, Accurate, and Label-Free Enumeration of Viable Microorganisms Enabled by Custom-Built On-Glass-Slide Culturing Device and Microscopic Scanning Song, Donghui Liu, Haomin Dong, Qiuchen Bian, Zichao Wu, Huixiang Lei, Yu Sensors (Basel) Article Accurately measuring the number of viable microorganisms plays an essential role in microbiological studies. Since the conventional agar method of enumerating visible colonies is time-consuming and not accurate, efforts have been made towards overcoming these limitations by counting the invisible micro-colonies. However, none of studies on micro-colony counting was able to save significant time or provide accurate results. Herein, we developed an on-glass-slide cell culture device that enables rapid formation of micro-colonies on a 0.38 mm-thick gel film without suffering from nutrient and oxygen deprivation during bacteria culturing. Employing a phase contrast imaging setup, we achieved rapid microscopic scanning of micro-colonies within a large sample area on the thin film without the need of fluorescent staining. Using Escherichia coli (E. coli) as a demonstration, our technique was able to shorten the culturing time to within 5 h and automatically enumerate the micro-colonies from the phase contrast images. Moreover, this method delivered more accurate counts than the conventional visible colony counting methods. Due to these advantages, this imaging-based micro-colony enumeration technique provides a new platform for the quantification of viable microorganisms. MDPI 2018-10-31 /pmc/articles/PMC6263752/ /pubmed/30384414 http://dx.doi.org/10.3390/s18113700 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Song, Donghui
Liu, Haomin
Dong, Qiuchen
Bian, Zichao
Wu, Huixiang
Lei, Yu
Digital, Rapid, Accurate, and Label-Free Enumeration of Viable Microorganisms Enabled by Custom-Built On-Glass-Slide Culturing Device and Microscopic Scanning
title Digital, Rapid, Accurate, and Label-Free Enumeration of Viable Microorganisms Enabled by Custom-Built On-Glass-Slide Culturing Device and Microscopic Scanning
title_full Digital, Rapid, Accurate, and Label-Free Enumeration of Viable Microorganisms Enabled by Custom-Built On-Glass-Slide Culturing Device and Microscopic Scanning
title_fullStr Digital, Rapid, Accurate, and Label-Free Enumeration of Viable Microorganisms Enabled by Custom-Built On-Glass-Slide Culturing Device and Microscopic Scanning
title_full_unstemmed Digital, Rapid, Accurate, and Label-Free Enumeration of Viable Microorganisms Enabled by Custom-Built On-Glass-Slide Culturing Device and Microscopic Scanning
title_short Digital, Rapid, Accurate, and Label-Free Enumeration of Viable Microorganisms Enabled by Custom-Built On-Glass-Slide Culturing Device and Microscopic Scanning
title_sort digital, rapid, accurate, and label-free enumeration of viable microorganisms enabled by custom-built on-glass-slide culturing device and microscopic scanning
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6263752/
https://www.ncbi.nlm.nih.gov/pubmed/30384414
http://dx.doi.org/10.3390/s18113700
work_keys_str_mv AT songdonghui digitalrapidaccurateandlabelfreeenumerationofviablemicroorganismsenabledbycustombuiltonglassslideculturingdeviceandmicroscopicscanning
AT liuhaomin digitalrapidaccurateandlabelfreeenumerationofviablemicroorganismsenabledbycustombuiltonglassslideculturingdeviceandmicroscopicscanning
AT dongqiuchen digitalrapidaccurateandlabelfreeenumerationofviablemicroorganismsenabledbycustombuiltonglassslideculturingdeviceandmicroscopicscanning
AT bianzichao digitalrapidaccurateandlabelfreeenumerationofviablemicroorganismsenabledbycustombuiltonglassslideculturingdeviceandmicroscopicscanning
AT wuhuixiang digitalrapidaccurateandlabelfreeenumerationofviablemicroorganismsenabledbycustombuiltonglassslideculturingdeviceandmicroscopicscanning
AT leiyu digitalrapidaccurateandlabelfreeenumerationofviablemicroorganismsenabledbycustombuiltonglassslideculturingdeviceandmicroscopicscanning