Evaluation of the derivates of phosphorescent Pt-coproporphyrin as intracellular oxygen-sensitive probes
Several new derivatives of the phosphorescent Pt(II)-coproporphyrin (PtCP) were evaluated with respect to the sensing of intracellular oxygen by phosphorescence quenching. Despite the more favorable molecular charge compared to PtCP, self-loading into mammalian cells was rather inefficient for all t...
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| Veröffentlicht in: | Analytical and bioanalytical chemistry 2010-03, Vol.396 (5), p.1793-1803 |
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| creator | Fercher, Andreas Ponomarev, Gelii V Yashunski, Dmitri Papkovsky, Dmitri |
| description | Several new derivatives of the phosphorescent Pt(II)-coproporphyrin (PtCP) were evaluated with respect to the sensing of intracellular oxygen by phosphorescence quenching. Despite the more favorable molecular charge compared to PtCP, self-loading into mammalian cells was rather inefficient for all the dyes, while cell loading by facilitated transport using transfection reagents produced promising results. The PtCP-NH₂ derivative, which gave best loading efficiency and S/N ratio, was investigated in detail including the optimisation of loading conditions, studies of sub-cellular localization, cytotoxicity, oxygen sensitivity and long-term signal stability. Being spectrally similar to the macromolecular MitoXpress™ probe currently used in this application, the PtCP-NH₂ demonstrated higher loading efficiency and phosphorescent signals, suitability for several problematic cell lines and a slightly increased lifetime scale for the physiological range (0-200 μM O₂). In physiological experiments with different cell types, mitochondrial uncouplers and inhibitors performed on a time-resolved fluorescence plate reader, this probe produced the anticipated profiles of intracellular oxygen concentration and responses to cell stimulation. Therefore, PtCP-NH₂ represents a convenient probe for the experiments and applications in which monitoring of cellular oxygen levels is required. |
| doi_str_mv | 10.1007/s00216-009-3399-z |
| format | Article |
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Despite the more favorable molecular charge compared to PtCP, self-loading into mammalian cells was rather inefficient for all the dyes, while cell loading by facilitated transport using transfection reagents produced promising results. The PtCP-NH₂ derivative, which gave best loading efficiency and S/N ratio, was investigated in detail including the optimisation of loading conditions, studies of sub-cellular localization, cytotoxicity, oxygen sensitivity and long-term signal stability. Being spectrally similar to the macromolecular MitoXpress™ probe currently used in this application, the PtCP-NH₂ demonstrated higher loading efficiency and phosphorescent signals, suitability for several problematic cell lines and a slightly increased lifetime scale for the physiological range (0-200 μM O₂). In physiological experiments with different cell types, mitochondrial uncouplers and inhibitors performed on a time-resolved fluorescence plate reader, this probe produced the anticipated profiles of intracellular oxygen concentration and responses to cell stimulation. Therefore, PtCP-NH₂ represents a convenient probe for the experiments and applications in which monitoring of cellular oxygen levels is required.</description><identifier>ISSN: 1618-2642</identifier><identifier>EISSN: 1618-2650</identifier><identifier>DOI: 10.1007/s00216-009-3399-z</identifier><identifier>PMID: 20063150</identifier><language>eng</language><publisher>Berlin/Heidelberg: Berlin/Heidelberg : Springer-Verlag</publisher><subject>Analytical Chemistry ; Animals ; Biochemistry ; Biosensing Techniques ; Calibration ; Cell metabolism and mitochondrial function ; Cell Survival ; Cells ; Cellular oxygen ; Characterization and Evaluation of Materials ; Chemistry ; Chemistry and Materials Science ; Comparative analysis ; Coproporphyrins - chemistry ; Fluorescent Dyes - chemical synthesis ; Fluorescent Dyes - chemistry ; Food Science ; Humans ; Infection ; Intracellular probe ; Investigations ; Laboratory Medicine ; Luminescent Measurements ; Mathematical analysis ; Molecular Probes - chemical synthesis ; Molecular Probes - chemistry ; Molecular Structure ; Monitoring/Environmental Analysis ; Organoplatinum Compounds - chemical synthesis ; Organoplatinum Compounds - chemistry ; Original Paper ; Oxygen - analysis ; Oxygen - metabolism ; Phosphorescent oxygen-sensitive probes ; Platinum ; Pt-porphyrins ; Rats ; Sensitivity and Specificity ; Time Factors ; Tumor Cells, Cultured</subject><ispartof>Analytical and bioanalytical chemistry, 2010-03, Vol.396 (5), p.1793-1803</ispartof><rights>Springer-Verlag 2010</rights><rights>COPYRIGHT 2010 Springer</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c565t-5deacfa3f4cb1e044f60222978c7ac9ab31994020f5d5249ea99d4cb4ae0ee373</citedby><cites>FETCH-LOGICAL-c565t-5deacfa3f4cb1e044f60222978c7ac9ab31994020f5d5249ea99d4cb4ae0ee373</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s00216-009-3399-z$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s00216-009-3399-z$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,780,784,27923,27924,41487,42556,51318</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/20063150$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Fercher, Andreas</creatorcontrib><creatorcontrib>Ponomarev, Gelii V</creatorcontrib><creatorcontrib>Yashunski, Dmitri</creatorcontrib><creatorcontrib>Papkovsky, Dmitri</creatorcontrib><title>Evaluation of the derivates of phosphorescent Pt-coproporphyrin as intracellular oxygen-sensitive probes</title><title>Analytical and bioanalytical chemistry</title><addtitle>Anal Bioanal Chem</addtitle><addtitle>Anal Bioanal Chem</addtitle><description>Several new derivatives of the phosphorescent Pt(II)-coproporphyrin (PtCP) were evaluated with respect to the sensing of intracellular oxygen by phosphorescence quenching. Despite the more favorable molecular charge compared to PtCP, self-loading into mammalian cells was rather inefficient for all the dyes, while cell loading by facilitated transport using transfection reagents produced promising results. The PtCP-NH₂ derivative, which gave best loading efficiency and S/N ratio, was investigated in detail including the optimisation of loading conditions, studies of sub-cellular localization, cytotoxicity, oxygen sensitivity and long-term signal stability. Being spectrally similar to the macromolecular MitoXpress™ probe currently used in this application, the PtCP-NH₂ demonstrated higher loading efficiency and phosphorescent signals, suitability for several problematic cell lines and a slightly increased lifetime scale for the physiological range (0-200 μM O₂). In physiological experiments with different cell types, mitochondrial uncouplers and inhibitors performed on a time-resolved fluorescence plate reader, this probe produced the anticipated profiles of intracellular oxygen concentration and responses to cell stimulation. Therefore, PtCP-NH₂ represents a convenient probe for the experiments and applications in which monitoring of cellular oxygen levels is required.</description><subject>Analytical Chemistry</subject><subject>Animals</subject><subject>Biochemistry</subject><subject>Biosensing Techniques</subject><subject>Calibration</subject><subject>Cell metabolism and mitochondrial function</subject><subject>Cell Survival</subject><subject>Cells</subject><subject>Cellular oxygen</subject><subject>Characterization and Evaluation of Materials</subject><subject>Chemistry</subject><subject>Chemistry and Materials Science</subject><subject>Comparative analysis</subject><subject>Coproporphyrins - chemistry</subject><subject>Fluorescent Dyes - chemical synthesis</subject><subject>Fluorescent Dyes - chemistry</subject><subject>Food Science</subject><subject>Humans</subject><subject>Infection</subject><subject>Intracellular probe</subject><subject>Investigations</subject><subject>Laboratory Medicine</subject><subject>Luminescent Measurements</subject><subject>Mathematical analysis</subject><subject>Molecular Probes - chemical synthesis</subject><subject>Molecular Probes - chemistry</subject><subject>Molecular Structure</subject><subject>Monitoring/Environmental Analysis</subject><subject>Organoplatinum Compounds - chemical synthesis</subject><subject>Organoplatinum Compounds - chemistry</subject><subject>Original Paper</subject><subject>Oxygen - analysis</subject><subject>Oxygen - metabolism</subject><subject>Phosphorescent oxygen-sensitive probes</subject><subject>Platinum</subject><subject>Pt-porphyrins</subject><subject>Rats</subject><subject>Sensitivity and Specificity</subject><subject>Time Factors</subject><subject>Tumor Cells, Cultured</subject><issn>1618-2642</issn><issn>1618-2650</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkl9rFDEUxQdRbK1-AF90wAd9mXrzdyaPpdRWKChon0M2c7ObMpusyczS7ac3w9SCIEVCSEh-5-aQe6rqLYFTAtB-zgCUyAZANYwp1dw_q46JJF1DpYDnj3tOj6pXOd8CENER-bI6ogCSEQHH1eZib4bJjD6GOrp63GDdY_J7M2KeD3abmMtMmC2Gsf4-NjbuUtzFtNsckg-1ybUPYzIWh2EaTKrj3WGNockYsh_9HuuCrzC_rl44M2R887CeVDdfLn6eXzXX3y6_np9dN1ZIMTaiR2OdYY7bFUHg3EmglKq2s62xyqwYUYoDBSd6QblCo1RfWG4QEFnLTqqPS93y7K8J86i3Ps_mTMA4Zd0KSVVHBfsPkknFpCCF_PQkSVrOpZSEz-iHBV2bAbUPLs5_M-P6rNju2rZ0oVCn_6DK6HHrbQzofDn_S0AWgU0x54RO75LfmnTQBPScBb1kQZcs6DkL-r5o3j24nlZb7B8Vf5pfALoAuVyFNSZ9G6cUSneerPp-ETkTtVknn_XND1osAumgU13HfgNqnskK</recordid><startdate>20100301</startdate><enddate>20100301</enddate><creator>Fercher, Andreas</creator><creator>Ponomarev, Gelii V</creator><creator>Yashunski, Dmitri</creator><creator>Papkovsky, Dmitri</creator><general>Berlin/Heidelberg : Springer-Verlag</general><general>Springer-Verlag</general><general>Springer</general><scope>FBQ</scope><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>7SR</scope><scope>7U5</scope><scope>8BQ</scope><scope>8FD</scope><scope>JG9</scope><scope>L7M</scope><scope>7QH</scope><scope>7UA</scope><scope>C1K</scope><scope>7X8</scope></search><sort><creationdate>20100301</creationdate><title>Evaluation of the derivates of phosphorescent Pt-coproporphyrin as intracellular oxygen-sensitive probes</title><author>Fercher, Andreas ; Ponomarev, Gelii V ; Yashunski, Dmitri ; Papkovsky, Dmitri</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c565t-5deacfa3f4cb1e044f60222978c7ac9ab31994020f5d5249ea99d4cb4ae0ee373</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>Analytical Chemistry</topic><topic>Animals</topic><topic>Biochemistry</topic><topic>Biosensing Techniques</topic><topic>Calibration</topic><topic>Cell metabolism and mitochondrial function</topic><topic>Cell Survival</topic><topic>Cells</topic><topic>Cellular oxygen</topic><topic>Characterization and Evaluation of Materials</topic><topic>Chemistry</topic><topic>Chemistry and Materials Science</topic><topic>Comparative analysis</topic><topic>Coproporphyrins - chemistry</topic><topic>Fluorescent Dyes - chemical synthesis</topic><topic>Fluorescent Dyes - chemistry</topic><topic>Food Science</topic><topic>Humans</topic><topic>Infection</topic><topic>Intracellular probe</topic><topic>Investigations</topic><topic>Laboratory Medicine</topic><topic>Luminescent Measurements</topic><topic>Mathematical analysis</topic><topic>Molecular Probes - chemical synthesis</topic><topic>Molecular Probes - chemistry</topic><topic>Molecular Structure</topic><topic>Monitoring/Environmental Analysis</topic><topic>Organoplatinum Compounds - chemical synthesis</topic><topic>Organoplatinum Compounds - chemistry</topic><topic>Original Paper</topic><topic>Oxygen - analysis</topic><topic>Oxygen - metabolism</topic><topic>Phosphorescent oxygen-sensitive probes</topic><topic>Platinum</topic><topic>Pt-porphyrins</topic><topic>Rats</topic><topic>Sensitivity and Specificity</topic><topic>Time Factors</topic><topic>Tumor Cells, Cultured</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Fercher, Andreas</creatorcontrib><creatorcontrib>Ponomarev, Gelii V</creatorcontrib><creatorcontrib>Yashunski, Dmitri</creatorcontrib><creatorcontrib>Papkovsky, Dmitri</creatorcontrib><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Engineered Materials Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>METADEX</collection><collection>Technology Research Database</collection><collection>Materials Research Database</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>Aqualine</collection><collection>Water Resources Abstracts</collection><collection>Environmental Sciences and Pollution Management</collection><collection>MEDLINE - Academic</collection><jtitle>Analytical and bioanalytical chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Fercher, Andreas</au><au>Ponomarev, Gelii V</au><au>Yashunski, Dmitri</au><au>Papkovsky, Dmitri</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Evaluation of the derivates of phosphorescent Pt-coproporphyrin as intracellular oxygen-sensitive probes</atitle><jtitle>Analytical and bioanalytical chemistry</jtitle><stitle>Anal Bioanal Chem</stitle><addtitle>Anal Bioanal Chem</addtitle><date>2010-03-01</date><risdate>2010</risdate><volume>396</volume><issue>5</issue><spage>1793</spage><epage>1803</epage><pages>1793-1803</pages><issn>1618-2642</issn><eissn>1618-2650</eissn><abstract>Several new derivatives of the phosphorescent Pt(II)-coproporphyrin (PtCP) were evaluated with respect to the sensing of intracellular oxygen by phosphorescence quenching. Despite the more favorable molecular charge compared to PtCP, self-loading into mammalian cells was rather inefficient for all the dyes, while cell loading by facilitated transport using transfection reagents produced promising results. The PtCP-NH₂ derivative, which gave best loading efficiency and S/N ratio, was investigated in detail including the optimisation of loading conditions, studies of sub-cellular localization, cytotoxicity, oxygen sensitivity and long-term signal stability. Being spectrally similar to the macromolecular MitoXpress™ probe currently used in this application, the PtCP-NH₂ demonstrated higher loading efficiency and phosphorescent signals, suitability for several problematic cell lines and a slightly increased lifetime scale for the physiological range (0-200 μM O₂). In physiological experiments with different cell types, mitochondrial uncouplers and inhibitors performed on a time-resolved fluorescence plate reader, this probe produced the anticipated profiles of intracellular oxygen concentration and responses to cell stimulation. Therefore, PtCP-NH₂ represents a convenient probe for the experiments and applications in which monitoring of cellular oxygen levels is required.</abstract><cop>Berlin/Heidelberg</cop><pub>Berlin/Heidelberg : Springer-Verlag</pub><pmid>20063150</pmid><doi>10.1007/s00216-009-3399-z</doi><tpages>11</tpages></addata></record> |
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| subjects | Analytical Chemistry Animals Biochemistry Biosensing Techniques Calibration Cell metabolism and mitochondrial function Cell Survival Cells Cellular oxygen Characterization and Evaluation of Materials Chemistry Chemistry and Materials Science Comparative analysis Coproporphyrins - chemistry Fluorescent Dyes - chemical synthesis Fluorescent Dyes - chemistry Food Science Humans Infection Intracellular probe Investigations Laboratory Medicine Luminescent Measurements Mathematical analysis Molecular Probes - chemical synthesis Molecular Probes - chemistry Molecular Structure Monitoring/Environmental Analysis Organoplatinum Compounds - chemical synthesis Organoplatinum Compounds - chemistry Original Paper Oxygen - analysis Oxygen - metabolism Phosphorescent oxygen-sensitive probes Platinum Pt-porphyrins Rats Sensitivity and Specificity Time Factors Tumor Cells, Cultured |
| title | Evaluation of the derivates of phosphorescent Pt-coproporphyrin as intracellular oxygen-sensitive probes |
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