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...
Autores principales: | , , , , , |
---|---|
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 |