A real-time method of imaging glucose uptake in single, living mammalian cells
This protocol details a method for monitoring glucose uptake into single, living mammalian cells using a fluorescent D -glucose derivative, 2-[ N -(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy- D -glucose (2-NBDG), as a tracer. The specifically designed chamber and superfusion system for evaluat...
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| Veröffentlicht in: | Nature protocols 2007-03, Vol.2 (3), p.753-762 |
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| creator | Matsuoka, Hideaki Inagaki, Nobuya Yamada, Katsuya Saito, Mikako |
| description | This protocol details a method for monitoring glucose uptake into single, living mammalian cells using a fluorescent
D
-glucose derivative, 2-[
N
-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-
D
-glucose (2-NBDG), as a tracer. The specifically designed chamber and superfusion system for evaluating 2-NBDG uptake into cells in real time can be combined with other fluorescent methods such as Ca
2+
imaging and the subsequent immunofluorescent classification of cells exhibiting divergent 2-NBDG uptake. The whole protocol, including immunocytochemistry, can be completed within 2 d (except for cell culture). The procedure for 2-NBDG synthesis is also presented. |
| doi_str_mv | 10.1038/nprot.2007.76 |
| format | Article |
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D
-glucose derivative, 2-[
N
-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-
D
-glucose (2-NBDG), as a tracer. The specifically designed chamber and superfusion system for evaluating 2-NBDG uptake into cells in real time can be combined with other fluorescent methods such as Ca
2+
imaging and the subsequent immunofluorescent classification of cells exhibiting divergent 2-NBDG uptake. The whole protocol, including immunocytochemistry, can be completed within 2 d (except for cell culture). The procedure for 2-NBDG synthesis is also presented.</description><identifier>ISSN: 1750-2799</identifier><identifier>ISSN: 1754-2189</identifier><identifier>EISSN: 1750-2799</identifier><identifier>DOI: 10.1038/nprot.2007.76</identifier><identifier>PMID: 17406637</identifier><language>eng</language><publisher>London: Nature Publishing Group UK</publisher><subject>4-Chloro-7-nitrobenzofurazan - analogs & derivatives ; 4-Chloro-7-nitrobenzofurazan - chemical synthesis ; 4-Chloro-7-nitrobenzofurazan - metabolism ; Analytical Chemistry ; Animals ; Biological Techniques ; Biological Transport - physiology ; Biomedical and Life Sciences ; Cell Line ; Computational Biology/Bioinformatics ; Decomposition ; Deoxyglucose - analogs & derivatives ; Deoxyglucose - chemical synthesis ; Deoxyglucose - metabolism ; Glucose ; Glucose - metabolism ; Glucose monitors ; Immunocytochemistry ; Immunohistochemistry ; Kinetics ; Life Sciences ; Mammals ; Medical research ; Methods ; Mice ; Microarrays ; Microscopy, Fluorescence ; Monitoring methods ; Organic Chemistry ; Physiological aspects ; Physiology ; Protocol ; Reagents ; Smooth muscle</subject><ispartof>Nature protocols, 2007-03, Vol.2 (3), p.753-762</ispartof><rights>Springer Nature Limited 2007</rights><rights>COPYRIGHT 2007 Nature Publishing Group</rights><rights>Copyright Nature Publishing Group Mar 2007</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c620t-88e09031d5277b37261c398c30aaf53e9ad18b3646c27e803f1b0817c1c9c7583</citedby><cites>FETCH-LOGICAL-c620t-88e09031d5277b37261c398c30aaf53e9ad18b3646c27e803f1b0817c1c9c7583</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1038/nprot.2007.76$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1038/nprot.2007.76$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,778,782,27911,27912,41475,42544,51306</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/17406637$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Matsuoka, Hideaki</creatorcontrib><creatorcontrib>Inagaki, Nobuya</creatorcontrib><creatorcontrib>Yamada, Katsuya</creatorcontrib><creatorcontrib>Saito, Mikako</creatorcontrib><title>A real-time method of imaging glucose uptake in single, living mammalian cells</title><title>Nature protocols</title><addtitle>Nat Protoc</addtitle><addtitle>Nat Protoc</addtitle><description>This protocol details a method for monitoring glucose uptake into single, living mammalian cells using a fluorescent
D
-glucose derivative, 2-[
N
-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-
D
-glucose (2-NBDG), as a tracer. The specifically designed chamber and superfusion system for evaluating 2-NBDG uptake into cells in real time can be combined with other fluorescent methods such as Ca
2+
imaging and the subsequent immunofluorescent classification of cells exhibiting divergent 2-NBDG uptake. The whole protocol, including immunocytochemistry, can be completed within 2 d (except for cell culture). The procedure for 2-NBDG synthesis is also presented.</description><subject>4-Chloro-7-nitrobenzofurazan - analogs & derivatives</subject><subject>4-Chloro-7-nitrobenzofurazan - chemical synthesis</subject><subject>4-Chloro-7-nitrobenzofurazan - metabolism</subject><subject>Analytical Chemistry</subject><subject>Animals</subject><subject>Biological Techniques</subject><subject>Biological Transport - physiology</subject><subject>Biomedical and Life Sciences</subject><subject>Cell Line</subject><subject>Computational Biology/Bioinformatics</subject><subject>Decomposition</subject><subject>Deoxyglucose - analogs & derivatives</subject><subject>Deoxyglucose - chemical synthesis</subject><subject>Deoxyglucose - metabolism</subject><subject>Glucose</subject><subject>Glucose - metabolism</subject><subject>Glucose monitors</subject><subject>Immunocytochemistry</subject><subject>Immunohistochemistry</subject><subject>Kinetics</subject><subject>Life Sciences</subject><subject>Mammals</subject><subject>Medical research</subject><subject>Methods</subject><subject>Mice</subject><subject>Microarrays</subject><subject>Microscopy, Fluorescence</subject><subject>Monitoring methods</subject><subject>Organic Chemistry</subject><subject>Physiological aspects</subject><subject>Physiology</subject><subject>Protocol</subject><subject>Reagents</subject><subject>Smooth muscle</subject><issn>1750-2799</issn><issn>1754-2189</issn><issn>1750-2799</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNqFktFr1TAUxosobk4ffZWAIAztNWnSJH28XKYOxgSd-Bhy09Oa2aTXJBX9703dxbvKRPKQcPLLx_m-nKJ4SvCKYCpf-10Y06rCWKwEv1ccE1HjshJNc__W-ah4FOM1xkxQLh4WR0QwzDkVx8XlGgXQQ5msA-QgfRlbNHbIOt1b36N-mMwYAU27pL8Csh7FXB7gFRrs9xlw2jk9WO2RgWGIj4sHnR4iPNnvJ8WnN2dXm3flxfu355v1RWl4hVMpJeAGU9LWlRBbKipODG2koVjrrqbQ6JbILeWMm0qAxLQjWyyJMMQ0RtSSnhQvbnSz-28TxKScjXMH2sM4RSUwZayR_wer3AVhuM7g87_A63EKPptQOWdWUyYkPVC9HkBZ340paDNLqjWRFWY1YSxTqzuovFpw1oweOpvriweniweZSfAj9XqKUZ1__LBkX_6bXV993lwu6fKGNmGMMUCndiF_bviZXc3GpPo9QGoeICV45p_tY5i2DtoDvZ-Yg7eYr3wP4XZOdyvuI_M6TQH-KC6pX-9J1-M</recordid><startdate>200703</startdate><enddate>200703</enddate><creator>Matsuoka, Hideaki</creator><creator>Inagaki, Nobuya</creator><creator>Yamada, Katsuya</creator><creator>Saito, Mikako</creator><general>Nature Publishing Group UK</general><general>Nature Publishing Group</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>ATWCN</scope><scope>ISR</scope><scope>3V.</scope><scope>7QG</scope><scope>7T5</scope><scope>7T7</scope><scope>7TM</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8FD</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>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>P64</scope><scope>PATMY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PYCSY</scope><scope>RC3</scope><scope>7QO</scope><scope>7X8</scope></search><sort><creationdate>200703</creationdate><title>A real-time method of imaging glucose uptake in single, living mammalian cells</title><author>Matsuoka, Hideaki ; Inagaki, Nobuya ; Yamada, Katsuya ; Saito, Mikako</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c620t-88e09031d5277b37261c398c30aaf53e9ad18b3646c27e803f1b0817c1c9c7583</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2007</creationdate><topic>4-Chloro-7-nitrobenzofurazan - analogs & derivatives</topic><topic>4-Chloro-7-nitrobenzofurazan - chemical synthesis</topic><topic>4-Chloro-7-nitrobenzofurazan - metabolism</topic><topic>Analytical Chemistry</topic><topic>Animals</topic><topic>Biological Techniques</topic><topic>Biological Transport - physiology</topic><topic>Biomedical and Life Sciences</topic><topic>Cell Line</topic><topic>Computational Biology/Bioinformatics</topic><topic>Decomposition</topic><topic>Deoxyglucose - analogs & derivatives</topic><topic>Deoxyglucose - chemical synthesis</topic><topic>Deoxyglucose - metabolism</topic><topic>Glucose</topic><topic>Glucose - metabolism</topic><topic>Glucose monitors</topic><topic>Immunocytochemistry</topic><topic>Immunohistochemistry</topic><topic>Kinetics</topic><topic>Life Sciences</topic><topic>Mammals</topic><topic>Medical research</topic><topic>Methods</topic><topic>Mice</topic><topic>Microarrays</topic><topic>Microscopy, Fluorescence</topic><topic>Monitoring methods</topic><topic>Organic Chemistry</topic><topic>Physiological aspects</topic><topic>Physiology</topic><topic>Protocol</topic><topic>Reagents</topic><topic>Smooth muscle</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Matsuoka, Hideaki</creatorcontrib><creatorcontrib>Inagaki, Nobuya</creatorcontrib><creatorcontrib>Yamada, Katsuya</creatorcontrib><creatorcontrib>Saito, Mikako</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Gale In Context: Middle School</collection><collection>Gale In Context: Science</collection><collection>ProQuest Central (Corporate)</collection><collection>Animal Behavior Abstracts</collection><collection>Immunology Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Nucleic Acids Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Technology Research Database</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>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection (ProQuest)</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</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>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Environmental Science 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>Environmental Science Collection</collection><collection>Genetics Abstracts</collection><collection>Biotechnology Research Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Nature protocols</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Matsuoka, Hideaki</au><au>Inagaki, Nobuya</au><au>Yamada, Katsuya</au><au>Saito, Mikako</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A real-time method of imaging glucose uptake in single, living mammalian cells</atitle><jtitle>Nature protocols</jtitle><stitle>Nat Protoc</stitle><addtitle>Nat Protoc</addtitle><date>2007-03</date><risdate>2007</risdate><volume>2</volume><issue>3</issue><spage>753</spage><epage>762</epage><pages>753-762</pages><issn>1750-2799</issn><issn>1754-2189</issn><eissn>1750-2799</eissn><abstract>This protocol details a method for monitoring glucose uptake into single, living mammalian cells using a fluorescent
D
-glucose derivative, 2-[
N
-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-
D
-glucose (2-NBDG), as a tracer. The specifically designed chamber and superfusion system for evaluating 2-NBDG uptake into cells in real time can be combined with other fluorescent methods such as Ca
2+
imaging and the subsequent immunofluorescent classification of cells exhibiting divergent 2-NBDG uptake. The whole protocol, including immunocytochemistry, can be completed within 2 d (except for cell culture). The procedure for 2-NBDG synthesis is also presented.</abstract><cop>London</cop><pub>Nature Publishing Group UK</pub><pmid>17406637</pmid><doi>10.1038/nprot.2007.76</doi><tpages>10</tpages></addata></record> |
| fulltext | fulltext |
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| subjects | 4-Chloro-7-nitrobenzofurazan - analogs & derivatives 4-Chloro-7-nitrobenzofurazan - chemical synthesis 4-Chloro-7-nitrobenzofurazan - metabolism Analytical Chemistry Animals Biological Techniques Biological Transport - physiology Biomedical and Life Sciences Cell Line Computational Biology/Bioinformatics Decomposition Deoxyglucose - analogs & derivatives Deoxyglucose - chemical synthesis Deoxyglucose - metabolism Glucose Glucose - metabolism Glucose monitors Immunocytochemistry Immunohistochemistry Kinetics Life Sciences Mammals Medical research Methods Mice Microarrays Microscopy, Fluorescence Monitoring methods Organic Chemistry Physiological aspects Physiology Protocol Reagents Smooth muscle |
| title | A real-time method of imaging glucose uptake in single, living mammalian cells |
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