Practical Implementation, Characterization and Applications of a Multi-Colour Time-Gated Luminescence Microscope
Time-gated luminescence microscopy using long-lifetime molecular probes can effectively eliminate autofluorescence to enable high contrast imaging. Here we investigate a new strategy of time-gated imaging for simultaneous visualisation of multiple species of microorganisms stained with long-lived co...
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| Veröffentlicht in: | Scientific reports 2014-10, Vol.4 (1), p.6597-6597, Article 6597 |
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| creator | Zhang, Lixin Zheng, Xianlin Deng, Wei Lu, Yiqing Lechevallier, Severine Ye, Zhiqiang Goldys, Ewa M. Dawes, Judith M. Piper, James A. Yuan, Jingli Verelst, Marc Jin, Dayong |
| description | Time-gated luminescence microscopy using long-lifetime molecular probes can effectively eliminate autofluorescence to enable high contrast imaging. Here we investigate a new strategy of time-gated imaging for simultaneous visualisation of multiple species of microorganisms stained with long-lived complexes under low-background conditions. This is realized by imaging two pathogenic organisms (
Giardia lamblia
stained with a red europium probe and
Cryptosporidium parvum
with a green terbium probe) at UV wavelengths (320–400 nm) through synchronization of a flash lamp with high repetition rate (1 kHz) to a robust time-gating detection unit. This approach provides four times enhancement in signal-to-background ratio over non-time-gated imaging, while the average signal intensity also increases six-fold compared with that under UV LED excitation. The high sensitivity is further confirmed by imaging the single europium-doped Y
2
O
2
S nanocrystals (150 nm). We report technical details regarding the time-gating detection unit and demonstrate its compatibility with commercial epi-fluorescence microscopes, providing a valuable and convenient addition to standard laboratory equipment. |
| doi_str_mv | 10.1038/srep06597 |
| format | Article |
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Giardia lamblia
stained with a red europium probe and
Cryptosporidium parvum
with a green terbium probe) at UV wavelengths (320–400 nm) through synchronization of a flash lamp with high repetition rate (1 kHz) to a robust time-gating detection unit. This approach provides four times enhancement in signal-to-background ratio over non-time-gated imaging, while the average signal intensity also increases six-fold compared with that under UV LED excitation. The high sensitivity is further confirmed by imaging the single europium-doped Y
2
O
2
S nanocrystals (150 nm). We report technical details regarding the time-gating detection unit and demonstrate its compatibility with commercial epi-fluorescence microscopes, providing a valuable and convenient addition to standard laboratory equipment.</description><identifier>ISSN: 2045-2322</identifier><identifier>EISSN: 2045-2322</identifier><identifier>DOI: 10.1038/srep06597</identifier><identifier>PMID: 25307702</identifier><language>eng</language><publisher>London: Nature Publishing Group UK</publisher><subject>639/624 ; 639/766/930/2735 ; Cryptosporidium ; Cryptosporidium parvum - ultrastructure ; Crystals ; Europium ; Europium - chemistry ; Gating ; Humanities and Social Sciences ; Luminescence ; Luminescent Measurements ; Microorganisms ; Microscopes ; Microscopy - methods ; Microscopy, Fluorescence - methods ; Molecular Imaging ; multidisciplinary ; Physics ; Probes ; Protozoa ; Science ; Synchronization ; Terbium ; Wavelengths</subject><ispartof>Scientific reports, 2014-10, Vol.4 (1), p.6597-6597, Article 6597</ispartof><rights>The Author(s) 2014</rights><rights>Copyright Nature Publishing Group Oct 2014</rights><rights>Distributed under a Creative Commons Attribution 4.0 International License</rights><rights>Copyright © 2014, Macmillan Publishers Limited. All rights reserved 2014 Macmillan Publishers Limited. All rights reserved</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c472t-4e49f91291e46084ed69f989d8a58c4228ba4615bdb2586686d62d94433b02e73</citedby><cites>FETCH-LOGICAL-c472t-4e49f91291e46084ed69f989d8a58c4228ba4615bdb2586686d62d94433b02e73</cites><orcidid>0000-0001-8005-5391</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4194433/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4194433/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,860,881,27901,27902,41096,42165,51551,53766,53768</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/25307702$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://hal.science/hal-01760884$$DView record in HAL$$Hfree_for_read</backlink></links><search><creatorcontrib>Zhang, Lixin</creatorcontrib><creatorcontrib>Zheng, Xianlin</creatorcontrib><creatorcontrib>Deng, Wei</creatorcontrib><creatorcontrib>Lu, Yiqing</creatorcontrib><creatorcontrib>Lechevallier, Severine</creatorcontrib><creatorcontrib>Ye, Zhiqiang</creatorcontrib><creatorcontrib>Goldys, Ewa M.</creatorcontrib><creatorcontrib>Dawes, Judith M.</creatorcontrib><creatorcontrib>Piper, James A.</creatorcontrib><creatorcontrib>Yuan, Jingli</creatorcontrib><creatorcontrib>Verelst, Marc</creatorcontrib><creatorcontrib>Jin, Dayong</creatorcontrib><title>Practical Implementation, Characterization and Applications of a Multi-Colour Time-Gated Luminescence Microscope</title><title>Scientific reports</title><addtitle>Sci Rep</addtitle><addtitle>Sci Rep</addtitle><description>Time-gated luminescence microscopy using long-lifetime molecular probes can effectively eliminate autofluorescence to enable high contrast imaging. Here we investigate a new strategy of time-gated imaging for simultaneous visualisation of multiple species of microorganisms stained with long-lived complexes under low-background conditions. This is realized by imaging two pathogenic organisms (
Giardia lamblia
stained with a red europium probe and
Cryptosporidium parvum
with a green terbium probe) at UV wavelengths (320–400 nm) through synchronization of a flash lamp with high repetition rate (1 kHz) to a robust time-gating detection unit. This approach provides four times enhancement in signal-to-background ratio over non-time-gated imaging, while the average signal intensity also increases six-fold compared with that under UV LED excitation. The high sensitivity is further confirmed by imaging the single europium-doped Y
2
O
2
S nanocrystals (150 nm). We report technical details regarding the time-gating detection unit and demonstrate its compatibility with commercial epi-fluorescence microscopes, providing a valuable and convenient addition to standard laboratory equipment.</description><subject>639/624</subject><subject>639/766/930/2735</subject><subject>Cryptosporidium</subject><subject>Cryptosporidium parvum - ultrastructure</subject><subject>Crystals</subject><subject>Europium</subject><subject>Europium - chemistry</subject><subject>Gating</subject><subject>Humanities and Social Sciences</subject><subject>Luminescence</subject><subject>Luminescent Measurements</subject><subject>Microorganisms</subject><subject>Microscopes</subject><subject>Microscopy - methods</subject><subject>Microscopy, Fluorescence - methods</subject><subject>Molecular Imaging</subject><subject>multidisciplinary</subject><subject>Physics</subject><subject>Probes</subject><subject>Protozoa</subject><subject>Science</subject><subject>Synchronization</subject><subject>Terbium</subject><subject>Wavelengths</subject><issn>2045-2322</issn><issn>2045-2322</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>C6C</sourceid><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><recordid>eNplUcFu1DAQtRAVrbY98APIEhdABGzHcexLpdUK2kpbwaGcLSeZ7bpy7GAnleDrcdiy3RZf7Jl5fvPmDUKvKflESSk_pwgDEZWqX6ATRnhVsJKxlwfvY3SW0h3Jp2KKU_UKHbOqJHVN2AkavkfTjrY1Dl_1g4Me_GhGG_xHvNqauQbR_v6bwcZ3eDkMLqPnOOGwwQZfT260xSq4MEV8Y3soLswIHV5PvfWQWvAt4GvbxpDaMMApOtoYl-Ds4V6gH1-_3Kwui_W3i6vVcl20vGZjwYGrjaJMUeCCSA6dyLFUnTSVbDljsjFc0KrpGlZJIaToBOsU52XZEAZ1uUDnO95hanrosowxGqeHaHsTf-lgrH5a8Xarb8O9zg7NLJng_Y5g--zb5XKt5xyhdVYm-T3N2HcPzWL4OUEadW_z5M4ZD2FKmgqaFUuWiRfo7TPoXTbOZys0lUrSihNOHpvPtuUNb_YKKNHz2vV-7Rn75nDSPfLfkjPgww6QcsnfQjxo-R_bH_qitk8</recordid><startdate>20141013</startdate><enddate>20141013</enddate><creator>Zhang, Lixin</creator><creator>Zheng, Xianlin</creator><creator>Deng, Wei</creator><creator>Lu, Yiqing</creator><creator>Lechevallier, Severine</creator><creator>Ye, Zhiqiang</creator><creator>Goldys, Ewa M.</creator><creator>Dawes, Judith M.</creator><creator>Piper, James A.</creator><creator>Yuan, Jingli</creator><creator>Verelst, Marc</creator><creator>Jin, Dayong</creator><general>Nature Publishing Group UK</general><general>Nature Publishing Group</general><scope>C6C</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>3V.</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>88I</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7P</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>7X8</scope><scope>1XC</scope><scope>VOOES</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0001-8005-5391</orcidid></search><sort><creationdate>20141013</creationdate><title>Practical Implementation, Characterization and Applications of a Multi-Colour Time-Gated Luminescence Microscope</title><author>Zhang, Lixin ; Zheng, Xianlin ; Deng, Wei ; Lu, Yiqing ; Lechevallier, Severine ; Ye, Zhiqiang ; Goldys, Ewa M. ; Dawes, Judith M. ; Piper, James A. ; Yuan, Jingli ; Verelst, Marc ; Jin, Dayong</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c472t-4e49f91291e46084ed69f989d8a58c4228ba4615bdb2586686d62d94433b02e73</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>639/624</topic><topic>639/766/930/2735</topic><topic>Cryptosporidium</topic><topic>Cryptosporidium parvum - ultrastructure</topic><topic>Crystals</topic><topic>Europium</topic><topic>Europium - chemistry</topic><topic>Gating</topic><topic>Humanities and Social Sciences</topic><topic>Luminescence</topic><topic>Luminescent Measurements</topic><topic>Microorganisms</topic><topic>Microscopes</topic><topic>Microscopy - methods</topic><topic>Microscopy, Fluorescence - methods</topic><topic>Molecular Imaging</topic><topic>multidisciplinary</topic><topic>Physics</topic><topic>Probes</topic><topic>Protozoa</topic><topic>Science</topic><topic>Synchronization</topic><topic>Terbium</topic><topic>Wavelengths</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Zhang, Lixin</creatorcontrib><creatorcontrib>Zheng, Xianlin</creatorcontrib><creatorcontrib>Deng, Wei</creatorcontrib><creatorcontrib>Lu, Yiqing</creatorcontrib><creatorcontrib>Lechevallier, Severine</creatorcontrib><creatorcontrib>Ye, Zhiqiang</creatorcontrib><creatorcontrib>Goldys, Ewa M.</creatorcontrib><creatorcontrib>Dawes, Judith M.</creatorcontrib><creatorcontrib>Piper, James A.</creatorcontrib><creatorcontrib>Yuan, Jingli</creatorcontrib><creatorcontrib>Verelst, Marc</creatorcontrib><creatorcontrib>Jin, Dayong</creatorcontrib><collection>Springer Nature OA Free Journals</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Science Database (Alumni Edition)</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Science Database</collection><collection>Biological Science Database</collection><collection>Publicly Available Content Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central Basic</collection><collection>MEDLINE - Academic</collection><collection>Hyper Article en Ligne (HAL)</collection><collection>Hyper Article en Ligne (HAL) (Open Access)</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Scientific reports</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Zhang, Lixin</au><au>Zheng, Xianlin</au><au>Deng, Wei</au><au>Lu, Yiqing</au><au>Lechevallier, Severine</au><au>Ye, Zhiqiang</au><au>Goldys, Ewa M.</au><au>Dawes, Judith M.</au><au>Piper, James A.</au><au>Yuan, Jingli</au><au>Verelst, Marc</au><au>Jin, Dayong</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Practical Implementation, Characterization and Applications of a Multi-Colour Time-Gated Luminescence Microscope</atitle><jtitle>Scientific reports</jtitle><stitle>Sci Rep</stitle><addtitle>Sci Rep</addtitle><date>2014-10-13</date><risdate>2014</risdate><volume>4</volume><issue>1</issue><spage>6597</spage><epage>6597</epage><pages>6597-6597</pages><artnum>6597</artnum><issn>2045-2322</issn><eissn>2045-2322</eissn><abstract>Time-gated luminescence microscopy using long-lifetime molecular probes can effectively eliminate autofluorescence to enable high contrast imaging. Here we investigate a new strategy of time-gated imaging for simultaneous visualisation of multiple species of microorganisms stained with long-lived complexes under low-background conditions. This is realized by imaging two pathogenic organisms (
Giardia lamblia
stained with a red europium probe and
Cryptosporidium parvum
with a green terbium probe) at UV wavelengths (320–400 nm) through synchronization of a flash lamp with high repetition rate (1 kHz) to a robust time-gating detection unit. This approach provides four times enhancement in signal-to-background ratio over non-time-gated imaging, while the average signal intensity also increases six-fold compared with that under UV LED excitation. The high sensitivity is further confirmed by imaging the single europium-doped Y
2
O
2
S nanocrystals (150 nm). We report technical details regarding the time-gating detection unit and demonstrate its compatibility with commercial epi-fluorescence microscopes, providing a valuable and convenient addition to standard laboratory equipment.</abstract><cop>London</cop><pub>Nature Publishing Group UK</pub><pmid>25307702</pmid><doi>10.1038/srep06597</doi><tpages>1</tpages><orcidid>https://orcid.org/0000-0001-8005-5391</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | 639/624 639/766/930/2735 Cryptosporidium Cryptosporidium parvum - ultrastructure Crystals Europium Europium - chemistry Gating Humanities and Social Sciences Luminescence Luminescent Measurements Microorganisms Microscopes Microscopy - methods Microscopy, Fluorescence - methods Molecular Imaging multidisciplinary Physics Probes Protozoa Science Synchronization Terbium Wavelengths |
| title | Practical Implementation, Characterization and Applications of a Multi-Colour Time-Gated Luminescence Microscope |
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