Bacterial and Mineral Elements in an Arctic Biofilm: A Correlative Study Using Fluorescence and Electron Microscopy
Abstract Few simple labeling methods exist for simultaneous fluorescence and electron microscopy of bacteria and biofilms. Here we describe the synthesis, characterization, and application of fluorescent nanoparticle quantum dot (QD) conjugates to target microbial species, including difficult to lab...
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croxfordunivpr:10.1017/s1431927609991334 2023-09-05T13:17:10+02:00 Bacterial and Mineral Elements in an Arctic Biofilm: A Correlative Study Using Fluorescence and Electron Microscopy Clarke, Samuel Mielke, Randall E. Neal, Andrea Holden, Patricia Nadeau, Jay L. 2010 http://dx.doi.org/10.1017/s1431927609991334 https://www.cambridge.org/core/services/aop-cambridge-core/content/view/S1431927609991334 en eng Oxford University Press (OUP) https://www.cambridge.org/core/terms Microscopy and Microanalysis volume 16, issue 2, page 153-165 ISSN 1431-9276 1435-8115 Instrumentation journal-article 2010 croxfordunivpr https://doi.org/10.1017/s1431927609991334 2023-08-18T09:59:37Z Abstract Few simple labeling methods exist for simultaneous fluorescence and electron microscopy of bacteria and biofilms. Here we describe the synthesis, characterization, and application of fluorescent nanoparticle quantum dot (QD) conjugates to target microbial species, including difficult to label Gram-negative strains. These QD conjugates impart contrast for both environmental scanning electron microscopy (ESEM) and fluorescence microscopy, permitting observation of living and fixed bacteria and biofilms. We apply these probes for studying biofilms extracted from perennial cold springs in the Canadian High Arctic, which is a particularly challenging system. In these biofilms, sulfur-metabolizing bacteria live in close association with unusual sulfur mineral formations. Following simple labeling protocols with the QD conjugates, we are able to image these organisms in fully-hydrated samples and visualize their relationship to the sulfur minerals using both ESEM and fluorescence microscopy. We then use scanning transmission electron microscopy to observe precipitated sulfur around individual cells and within the biofilm lattice. All combined, this information sheds light on the possible mechanisms of biofilm and mineral structure formation. These new QD conjugates and techniques are highly transferable to many other microbiological applications, especially those involving Gram-negative bacteria, and can be used for correlated fluorescence and electron microscopy. Article in Journal/Newspaper Arctic Oxford University Press (via Crossref) Arctic Microscopy and Microanalysis 16 2 153 165 |
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Instrumentation Clarke, Samuel Mielke, Randall E. Neal, Andrea Holden, Patricia Nadeau, Jay L. Bacterial and Mineral Elements in an Arctic Biofilm: A Correlative Study Using Fluorescence and Electron Microscopy |
topic_facet |
Instrumentation |
description |
Abstract Few simple labeling methods exist for simultaneous fluorescence and electron microscopy of bacteria and biofilms. Here we describe the synthesis, characterization, and application of fluorescent nanoparticle quantum dot (QD) conjugates to target microbial species, including difficult to label Gram-negative strains. These QD conjugates impart contrast for both environmental scanning electron microscopy (ESEM) and fluorescence microscopy, permitting observation of living and fixed bacteria and biofilms. We apply these probes for studying biofilms extracted from perennial cold springs in the Canadian High Arctic, which is a particularly challenging system. In these biofilms, sulfur-metabolizing bacteria live in close association with unusual sulfur mineral formations. Following simple labeling protocols with the QD conjugates, we are able to image these organisms in fully-hydrated samples and visualize their relationship to the sulfur minerals using both ESEM and fluorescence microscopy. We then use scanning transmission electron microscopy to observe precipitated sulfur around individual cells and within the biofilm lattice. All combined, this information sheds light on the possible mechanisms of biofilm and mineral structure formation. These new QD conjugates and techniques are highly transferable to many other microbiological applications, especially those involving Gram-negative bacteria, and can be used for correlated fluorescence and electron microscopy. |
format |
Article in Journal/Newspaper |
author |
Clarke, Samuel Mielke, Randall E. Neal, Andrea Holden, Patricia Nadeau, Jay L. |
author_facet |
Clarke, Samuel Mielke, Randall E. Neal, Andrea Holden, Patricia Nadeau, Jay L. |
author_sort |
Clarke, Samuel |
title |
Bacterial and Mineral Elements in an Arctic Biofilm: A Correlative Study Using Fluorescence and Electron Microscopy |
title_short |
Bacterial and Mineral Elements in an Arctic Biofilm: A Correlative Study Using Fluorescence and Electron Microscopy |
title_full |
Bacterial and Mineral Elements in an Arctic Biofilm: A Correlative Study Using Fluorescence and Electron Microscopy |
title_fullStr |
Bacterial and Mineral Elements in an Arctic Biofilm: A Correlative Study Using Fluorescence and Electron Microscopy |
title_full_unstemmed |
Bacterial and Mineral Elements in an Arctic Biofilm: A Correlative Study Using Fluorescence and Electron Microscopy |
title_sort |
bacterial and mineral elements in an arctic biofilm: a correlative study using fluorescence and electron microscopy |
publisher |
Oxford University Press (OUP) |
publishDate |
2010 |
url |
http://dx.doi.org/10.1017/s1431927609991334 https://www.cambridge.org/core/services/aop-cambridge-core/content/view/S1431927609991334 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Microscopy and Microanalysis volume 16, issue 2, page 153-165 ISSN 1431-9276 1435-8115 |
op_rights |
https://www.cambridge.org/core/terms |
op_doi |
https://doi.org/10.1017/s1431927609991334 |
container_title |
Microscopy and Microanalysis |
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16 |
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2 |
container_start_page |
153 |
op_container_end_page |
165 |
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1776198449875648512 |