Document Detail


Probing single molecules in single living cells.
MedLine Citation:
PMID:  11101238     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Single-molecule detection in single living cells has been achieved by using confocal fluorescence microscopy and externally tagged probe molecules. The intracellular background fluorescence is substantially higher than that in aqueous buffer, but this background is continuous and stable and does not significantly interfere with the measurement of single-molecule photon bursts. As a result, single-molecule data have been obtained on three types of fluorescent probes at spatially resolved locations (e.g., cytoplasm and nucleus) inside human HeLa cells. First, the iron transport protein transferrin labeled with tetramethylrhodamine undergoes rapid receptor-mediated endocytosis, and single transferrin molecules are detected inside living cells. Second, the cationic dye rhodamine 6G (R6G) enters cultured cells by a potential-driven process, and single R6G molecules are observed as intense photon bursts when they move in and out of the intracellular laser beam. Third, we report results on synthetic oligonucleotides that are tagged with a fluorescent dye and are taken up by living cells via a passive, nonendocytic pathway.
Authors:
T A Byassee; W C Chan; S Nie
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Analytical chemistry     Volume:  72     ISSN:  0003-2700     ISO Abbreviation:  Anal. Chem.     Publication Date:  2000 Nov 
Date Detail:
Created Date:  2000-12-04     Completed Date:  2000-12-22     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0370536     Medline TA:  Anal Chem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  5606-11     Citation Subset:  IM    
Affiliation:
Department of Chemistry, Indiana University, Bloomington 47405, USA.
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MeSH Terms
Descriptor/Qualifier:
Hela Cells
Humans
Microscopy, Confocal / methods
Microscopy, Fluorescence / methods
Molecular Probes*
Chemical
Reg. No./Substance:
0/Molecular Probes

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