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Microfluidic Cultivation and Laser Tweezers Raman Spectroscopy of E. coli under Antibiotic Stress

Analyzing the cells in various body fluids can greatly deepen the understanding of the mechanisms governing the cellular physiology. Due to the variability of physiological and metabolic states, it is important to be able to perform such studies on individual cells. Therefore, we developed an optofl...

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Autores principales: Pilát, Zdeněk, Bernatová, Silvie, Ježek, Jan, Kirchhoff, Johanna, Tannert, Astrid, Neugebauer, Ute, Samek, Ota, Zemánek, Pavel
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5982924/
https://www.ncbi.nlm.nih.gov/pubmed/29783713
http://dx.doi.org/10.3390/s18051623
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author Pilát, Zdeněk
Bernatová, Silvie
Ježek, Jan
Kirchhoff, Johanna
Tannert, Astrid
Neugebauer, Ute
Samek, Ota
Zemánek, Pavel
author_facet Pilát, Zdeněk
Bernatová, Silvie
Ježek, Jan
Kirchhoff, Johanna
Tannert, Astrid
Neugebauer, Ute
Samek, Ota
Zemánek, Pavel
author_sort Pilát, Zdeněk
collection PubMed
description Analyzing the cells in various body fluids can greatly deepen the understanding of the mechanisms governing the cellular physiology. Due to the variability of physiological and metabolic states, it is important to be able to perform such studies on individual cells. Therefore, we developed an optofluidic system in which we precisely manipulated and monitored individual cells of Escherichia coli. We tested optical micromanipulation in a microfluidic chamber chip by transferring individual bacteria into the chambers. We then subjected the cells in the chambers to antibiotic cefotaxime and we observed the changes by using time-lapse microscopy. Separately, we used laser tweezers Raman spectroscopy (LTRS) in a different micro-chamber chip to manipulate and analyze individual cefotaxime-treated E. coli cells. Additionally, we performed conventional Raman micro-spectroscopic measurements of E. coli cells in a micro-chamber. We found observable changes in the cellular morphology (cell elongation) and in Raman spectra, which were consistent with other recently published observations. The principal component analysis (PCA) of Raman data distinguished between the cefotaxime treated cells and control. We tested the capabilities of the optofluidic system and found it to be a reliable and versatile solution for this class of microbiological experiments.
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spelling pubmed-59829242018-06-05 Microfluidic Cultivation and Laser Tweezers Raman Spectroscopy of E. coli under Antibiotic Stress Pilát, Zdeněk Bernatová, Silvie Ježek, Jan Kirchhoff, Johanna Tannert, Astrid Neugebauer, Ute Samek, Ota Zemánek, Pavel Sensors (Basel) Article Analyzing the cells in various body fluids can greatly deepen the understanding of the mechanisms governing the cellular physiology. Due to the variability of physiological and metabolic states, it is important to be able to perform such studies on individual cells. Therefore, we developed an optofluidic system in which we precisely manipulated and monitored individual cells of Escherichia coli. We tested optical micromanipulation in a microfluidic chamber chip by transferring individual bacteria into the chambers. We then subjected the cells in the chambers to antibiotic cefotaxime and we observed the changes by using time-lapse microscopy. Separately, we used laser tweezers Raman spectroscopy (LTRS) in a different micro-chamber chip to manipulate and analyze individual cefotaxime-treated E. coli cells. Additionally, we performed conventional Raman micro-spectroscopic measurements of E. coli cells in a micro-chamber. We found observable changes in the cellular morphology (cell elongation) and in Raman spectra, which were consistent with other recently published observations. The principal component analysis (PCA) of Raman data distinguished between the cefotaxime treated cells and control. We tested the capabilities of the optofluidic system and found it to be a reliable and versatile solution for this class of microbiological experiments. MDPI 2018-05-18 /pmc/articles/PMC5982924/ /pubmed/29783713 http://dx.doi.org/10.3390/s18051623 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
Pilát, Zdeněk
Bernatová, Silvie
Ježek, Jan
Kirchhoff, Johanna
Tannert, Astrid
Neugebauer, Ute
Samek, Ota
Zemánek, Pavel
Microfluidic Cultivation and Laser Tweezers Raman Spectroscopy of E. coli under Antibiotic Stress
title Microfluidic Cultivation and Laser Tweezers Raman Spectroscopy of E. coli under Antibiotic Stress
title_full Microfluidic Cultivation and Laser Tweezers Raman Spectroscopy of E. coli under Antibiotic Stress
title_fullStr Microfluidic Cultivation and Laser Tweezers Raman Spectroscopy of E. coli under Antibiotic Stress
title_full_unstemmed Microfluidic Cultivation and Laser Tweezers Raman Spectroscopy of E. coli under Antibiotic Stress
title_short Microfluidic Cultivation and Laser Tweezers Raman Spectroscopy of E. coli under Antibiotic Stress
title_sort microfluidic cultivation and laser tweezers raman spectroscopy of e. coli under antibiotic stress
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5982924/
https://www.ncbi.nlm.nih.gov/pubmed/29783713
http://dx.doi.org/10.3390/s18051623
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