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Optical Screening of Novel Bacteria-specific Probes on Ex Vivo Human Lung Tissue by Confocal Laser Endomicroscopy

Improving the speed and accuracy of bacterial detection is important for patient stratification and to ensure the appropriate use of antimicrobials. To achieve this goal, the development of diagnostic techniques to recognize bacterial presence in real-time at the point-of-care is required. Optical i...

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Autores principales: Mills, Bethany, Akram, Ahsan R., Scholefield, Emma, Bradley, Mark, Dhaliwal, Kevin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MyJove Corporation 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5755507/
https://www.ncbi.nlm.nih.gov/pubmed/29286374
http://dx.doi.org/10.3791/56284
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author Mills, Bethany
Akram, Ahsan R.
Scholefield, Emma
Bradley, Mark
Dhaliwal, Kevin
author_facet Mills, Bethany
Akram, Ahsan R.
Scholefield, Emma
Bradley, Mark
Dhaliwal, Kevin
author_sort Mills, Bethany
collection PubMed
description Improving the speed and accuracy of bacterial detection is important for patient stratification and to ensure the appropriate use of antimicrobials. To achieve this goal, the development of diagnostic techniques to recognize bacterial presence in real-time at the point-of-care is required. Optical imaging for direct identification of bacteria within the host is an attractive approach. Several attempts at chemical probe design and validation have been investigated, however none have yet been successfully translated into the clinic. Here we describe a method for ex vivo validation of bacteria-specific probes for identification of bacteria within the distal lung, imaged by fibered confocal fluorescence microscopy (FCFM). Our model used ex vivo human lung tissue and a clinically approved confocal laser endomicroscopy (CLE) platform to screen novel bacteria-specific imaging compounds, closely mimicking imaging conditions expected to be encountered with patients. Therefore, screening compounds by this technique provides confidence of potential clinical tractability.
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spelling pubmed-57555072018-01-19 Optical Screening of Novel Bacteria-specific Probes on Ex Vivo Human Lung Tissue by Confocal Laser Endomicroscopy Mills, Bethany Akram, Ahsan R. Scholefield, Emma Bradley, Mark Dhaliwal, Kevin J Vis Exp Immunology Improving the speed and accuracy of bacterial detection is important for patient stratification and to ensure the appropriate use of antimicrobials. To achieve this goal, the development of diagnostic techniques to recognize bacterial presence in real-time at the point-of-care is required. Optical imaging for direct identification of bacteria within the host is an attractive approach. Several attempts at chemical probe design and validation have been investigated, however none have yet been successfully translated into the clinic. Here we describe a method for ex vivo validation of bacteria-specific probes for identification of bacteria within the distal lung, imaged by fibered confocal fluorescence microscopy (FCFM). Our model used ex vivo human lung tissue and a clinically approved confocal laser endomicroscopy (CLE) platform to screen novel bacteria-specific imaging compounds, closely mimicking imaging conditions expected to be encountered with patients. Therefore, screening compounds by this technique provides confidence of potential clinical tractability. MyJove Corporation 2017-11-29 /pmc/articles/PMC5755507/ /pubmed/29286374 http://dx.doi.org/10.3791/56284 Text en Copyright © 2017, Journal of Visualized Experiments http://creativecommons.org/licenses/by/3.0/us/ This is an open-access article distributed under the terms of the Creative Commons Attribution 3.0 License. To view a copy of this license, visithttp://creativecommons.org/licenses/by/3.0/us/
spellingShingle Immunology
Mills, Bethany
Akram, Ahsan R.
Scholefield, Emma
Bradley, Mark
Dhaliwal, Kevin
Optical Screening of Novel Bacteria-specific Probes on Ex Vivo Human Lung Tissue by Confocal Laser Endomicroscopy
title Optical Screening of Novel Bacteria-specific Probes on Ex Vivo Human Lung Tissue by Confocal Laser Endomicroscopy
title_full Optical Screening of Novel Bacteria-specific Probes on Ex Vivo Human Lung Tissue by Confocal Laser Endomicroscopy
title_fullStr Optical Screening of Novel Bacteria-specific Probes on Ex Vivo Human Lung Tissue by Confocal Laser Endomicroscopy
title_full_unstemmed Optical Screening of Novel Bacteria-specific Probes on Ex Vivo Human Lung Tissue by Confocal Laser Endomicroscopy
title_short Optical Screening of Novel Bacteria-specific Probes on Ex Vivo Human Lung Tissue by Confocal Laser Endomicroscopy
title_sort optical screening of novel bacteria-specific probes on ex vivo human lung tissue by confocal laser endomicroscopy
topic Immunology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5755507/
https://www.ncbi.nlm.nih.gov/pubmed/29286374
http://dx.doi.org/10.3791/56284
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