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Susceptibility of the Different Oxygen-Sensing Probes to Interferences in Respirometric Bacterial Assays with Complex Media
Respirometric microbial assays are gaining popularity, but their uptake is limited by the availability of optimal O sensing materials and the challenge of validating assays with complex real samples. We conducted a comparative evaluation of four different O -sensing probes based on Pt-porphyrin phos...
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| Published in: | Sensors (Basel, Switzerland) Switzerland), 2024-01, Vol.24 (1), p.267 |
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| creator | Zanetti, Chiara Li, Liang Gaspar, Rafael Di Lazaro Santovito, Elisa Elisseeva, Sophia Collins, Stuart G Maguire, Anita R Papkovsky, Dmitri B |
| description | Respirometric microbial assays are gaining popularity, but their uptake is limited by the availability of optimal O
sensing materials and the challenge of validating assays with complex real samples. We conducted a comparative evaluation of four different O
-sensing probes based on Pt-porphyrin phosphors in respirometric bacterial assays performed on standard time-resolved fluorescence reader. The macromolecular MitoXpress, nanoparticle NanO2 and small molecule PtGlc
and PtPEG
probes were assessed with
cells in five growth media: nutrient broth (NB), McConkey (MC), Rapid Coliform ChromoSelect (RCC), M-Lauryl lauryl sulfate (MLS), and Minerals-Modified Glutamate (MMG) media. Respiration profiles of the cells were recorded and analyzed, along with densitometry profiles and quenching studies of individual media components. This revealed several limiting factors and interferences impacting assay performance, which include probe quenched lifetime, instrument temporal resolution, inner filter effects (mainly by indicator dyes), probe binding to lipophilic components, and dynamic and static quenching by media components. The study allowed for the ranking of the probes based on their ruggedness, resilience to interferences and overall performance in respirometric bacterial assays. The 'shielded' probe NanO2 outperformed the established MitoXpress probe and the small molecule probes PtGlc
and PtPEG
. |
| doi_str_mv | 10.3390/s24010267 |
| format | article |
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sensing materials and the challenge of validating assays with complex real samples. We conducted a comparative evaluation of four different O
-sensing probes based on Pt-porphyrin phosphors in respirometric bacterial assays performed on standard time-resolved fluorescence reader. The macromolecular MitoXpress, nanoparticle NanO2 and small molecule PtGlc
and PtPEG
probes were assessed with
cells in five growth media: nutrient broth (NB), McConkey (MC), Rapid Coliform ChromoSelect (RCC), M-Lauryl lauryl sulfate (MLS), and Minerals-Modified Glutamate (MMG) media. Respiration profiles of the cells were recorded and analyzed, along with densitometry profiles and quenching studies of individual media components. This revealed several limiting factors and interferences impacting assay performance, which include probe quenched lifetime, instrument temporal resolution, inner filter effects (mainly by indicator dyes), probe binding to lipophilic components, and dynamic and static quenching by media components. The study allowed for the ranking of the probes based on their ruggedness, resilience to interferences and overall performance in respirometric bacterial assays. The 'shielded' probe NanO2 outperformed the established MitoXpress probe and the small molecule probes PtGlc
and PtPEG
.</description><identifier>ISSN: 1424-8220</identifier><identifier>EISSN: 1424-8220</identifier><identifier>DOI: 10.3390/s24010267</identifier><identifier>PMID: 38203132</identifier><language>eng</language><publisher>Switzerland: MDPI AG</publisher><subject>Bacteria ; bacterial cell respiration ; Biological Assay ; Biological Transport ; Dyes ; E coli ; Environmental sciences ; Escherichia coli ; Glutamic Acid ; Microorganisms ; Mineral oils ; optical oxygen respirometry ; Optical properties ; Oxygen ; Plating ; quenched-phosphorescence oxygen sensing ; Respiration ; selective media ; Sensors ; Surfactants ; water-soluble oxygen probes</subject><ispartof>Sensors (Basel, Switzerland), 2024-01, Vol.24 (1), p.267</ispartof><rights>COPYRIGHT 2024 MDPI AG</rights><rights>2024 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 (https://creativecommons.org/licenses/by/4.0/). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2024 by the authors. 2024</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c509t-d2e77e3528625b168efffb3f010a7d2adf3a496853fec0a3c50886faa7e698553</citedby><cites>FETCH-LOGICAL-c509t-d2e77e3528625b168efffb3f010a7d2adf3a496853fec0a3c50886faa7e698553</cites><orcidid>0000-0003-1556-1145 ; 0000-0003-3756-403X</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.proquest.com/docview/2912796495/fulltextPDF?pq-origsite=primo$$EPDF$$P50$$Gproquest$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.proquest.com/docview/2912796495?pq-origsite=primo$$EHTML$$P50$$Gproquest$$Hfree_for_read</linktohtml><link.rule.ids>230,314,724,777,781,882,25734,27905,27906,36993,36994,44571,53772,53774,74875</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/38203132$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Zanetti, Chiara</creatorcontrib><creatorcontrib>Li, Liang</creatorcontrib><creatorcontrib>Gaspar, Rafael Di Lazaro</creatorcontrib><creatorcontrib>Santovito, Elisa</creatorcontrib><creatorcontrib>Elisseeva, Sophia</creatorcontrib><creatorcontrib>Collins, Stuart G</creatorcontrib><creatorcontrib>Maguire, Anita R</creatorcontrib><creatorcontrib>Papkovsky, Dmitri B</creatorcontrib><title>Susceptibility of the Different Oxygen-Sensing Probes to Interferences in Respirometric Bacterial Assays with Complex Media</title><title>Sensors (Basel, Switzerland)</title><addtitle>Sensors (Basel)</addtitle><description>Respirometric microbial assays are gaining popularity, but their uptake is limited by the availability of optimal O
sensing materials and the challenge of validating assays with complex real samples. We conducted a comparative evaluation of four different O
-sensing probes based on Pt-porphyrin phosphors in respirometric bacterial assays performed on standard time-resolved fluorescence reader. The macromolecular MitoXpress, nanoparticle NanO2 and small molecule PtGlc
and PtPEG
probes were assessed with
cells in five growth media: nutrient broth (NB), McConkey (MC), Rapid Coliform ChromoSelect (RCC), M-Lauryl lauryl sulfate (MLS), and Minerals-Modified Glutamate (MMG) media. Respiration profiles of the cells were recorded and analyzed, along with densitometry profiles and quenching studies of individual media components. This revealed several limiting factors and interferences impacting assay performance, which include probe quenched lifetime, instrument temporal resolution, inner filter effects (mainly by indicator dyes), probe binding to lipophilic components, and dynamic and static quenching by media components. The study allowed for the ranking of the probes based on their ruggedness, resilience to interferences and overall performance in respirometric bacterial assays. The 'shielded' probe NanO2 outperformed the established MitoXpress probe and the small molecule probes PtGlc
and PtPEG
.</description><subject>Bacteria</subject><subject>bacterial cell respiration</subject><subject>Biological Assay</subject><subject>Biological Transport</subject><subject>Dyes</subject><subject>E coli</subject><subject>Environmental sciences</subject><subject>Escherichia coli</subject><subject>Glutamic Acid</subject><subject>Microorganisms</subject><subject>Mineral oils</subject><subject>optical oxygen respirometry</subject><subject>Optical properties</subject><subject>Oxygen</subject><subject>Plating</subject><subject>quenched-phosphorescence oxygen sensing</subject><subject>Respiration</subject><subject>selective media</subject><subject>Sensors</subject><subject>Surfactants</subject><subject>water-soluble oxygen probes</subject><issn>1424-8220</issn><issn>1424-8220</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2024</creationdate><recordtype>article</recordtype><sourceid>PIMPY</sourceid><sourceid>DOA</sourceid><recordid>eNpdkk1vEzEQhlcIREvhwB9AlrjAYYu_1rs-oRC-IhUVUThbXu84cbRrB9uBRvx5nKZELfLB1swzr_2Op6qeE3zOmMRvEuWYYCraB9Up4ZTXHaX44Z3zSfUkpTXGlDHWPa5OWEcxI4yeVn-utsnAJrvejS7vULAorwC9d9ZCBJ_R5fVuCb6-Ap-cX6KvMfSQUA5o4TPEG8iUgPPoG6SNi2GCHJ1B77QpeadHNEtJ7xL67fIKzcO0GeEafYHB6afVI6vHBM9u97Pqx8cP3-ef64vLT4v57KI2DZa5Hii0LbCGdoI2PREdWGt7Zotl3Q5UD5ZpLkXXMAsGa1aquk5YrVsQsmsadlYtDrpD0Gu1iW7ScaeCduomEOJS6ZidGUH13AzDINpWasEJl1IwLVtBJDSaEcOL1tuD1mbbTzCY0qKox3ui9zPerdQy_FIEtx2hZK_w6lYhhp9bSFlNrnzBOGoPYZsUlYRxXtyIgr78D12HbfSlV3uKtlJwubd3fqCWujhw3oZysSlrgMmZ4MG6Ep8VT6whuCOl4PWhwMSQUgR7fD7Baj9Q6jhQhX1x1--R_DdB7C8wB8Za</recordid><startdate>20240101</startdate><enddate>20240101</enddate><creator>Zanetti, Chiara</creator><creator>Li, Liang</creator><creator>Gaspar, Rafael Di Lazaro</creator><creator>Santovito, Elisa</creator><creator>Elisseeva, Sophia</creator><creator>Collins, Stuart G</creator><creator>Maguire, Anita R</creator><creator>Papkovsky, Dmitri B</creator><general>MDPI AG</general><general>MDPI</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BENPR</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>K9.</scope><scope>M0S</scope><scope>M1P</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope><orcidid>https://orcid.org/0000-0003-1556-1145</orcidid><orcidid>https://orcid.org/0000-0003-3756-403X</orcidid></search><sort><creationdate>20240101</creationdate><title>Susceptibility of the Different Oxygen-Sensing Probes to Interferences in Respirometric Bacterial Assays with Complex Media</title><author>Zanetti, Chiara ; 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sensing materials and the challenge of validating assays with complex real samples. We conducted a comparative evaluation of four different O
-sensing probes based on Pt-porphyrin phosphors in respirometric bacterial assays performed on standard time-resolved fluorescence reader. The macromolecular MitoXpress, nanoparticle NanO2 and small molecule PtGlc
and PtPEG
probes were assessed with
cells in five growth media: nutrient broth (NB), McConkey (MC), Rapid Coliform ChromoSelect (RCC), M-Lauryl lauryl sulfate (MLS), and Minerals-Modified Glutamate (MMG) media. Respiration profiles of the cells were recorded and analyzed, along with densitometry profiles and quenching studies of individual media components. This revealed several limiting factors and interferences impacting assay performance, which include probe quenched lifetime, instrument temporal resolution, inner filter effects (mainly by indicator dyes), probe binding to lipophilic components, and dynamic and static quenching by media components. The study allowed for the ranking of the probes based on their ruggedness, resilience to interferences and overall performance in respirometric bacterial assays. The 'shielded' probe NanO2 outperformed the established MitoXpress probe and the small molecule probes PtGlc
and PtPEG
.</abstract><cop>Switzerland</cop><pub>MDPI AG</pub><pmid>38203132</pmid><doi>10.3390/s24010267</doi><orcidid>https://orcid.org/0000-0003-1556-1145</orcidid><orcidid>https://orcid.org/0000-0003-3756-403X</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | Bacteria bacterial cell respiration Biological Assay Biological Transport Dyes E coli Environmental sciences Escherichia coli Glutamic Acid Microorganisms Mineral oils optical oxygen respirometry Optical properties Oxygen Plating quenched-phosphorescence oxygen sensing Respiration selective media Sensors Surfactants water-soluble oxygen probes |
| title | Susceptibility of the Different Oxygen-Sensing Probes to Interferences in Respirometric Bacterial Assays with Complex Media |
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