| A hyphenated optical trap capillary electrophoresis laser induced native fluorescence system for single-cell chemical analysis. | |
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MedLine Citation:
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PMID: 22543409 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Single-cell measurements allow a unique glimpse into cell-to-cell heterogeneity; even small changes in selected cells can have a profound impact on an organism's physiology. Here an integrated approach to single-cell chemical sampling and assay are described. Capillary electrophoresis (CE) with laser-induced native fluorescence (LINF) has the sensitivity to characterize natively fluorescent indoles and catechols within individual cells. While the separation and detection approaches are well established, the sampling and injection of individually selected cells requires new approaches. We describe an optimized system that interfaces a single-beam optical trap with CE and multichannel LINF detection. A cell is localized within the trap and then the capillary inlet is positioned near the cell using a computer-controlled micromanipulator. Hydrodynamic injection allows cell lysis to occur within the capillary inlet, followed by the CE separation and LINF detection. The use of multiple emission wavelengths allows improved analyte identification based on differences in analyte fluorescence emission profiles and migration time. The system enables injections of individual rat pinealocytes and quantification of their endogenous indoles, including serotonin, N-acetyl-serotonin, 5-hydroxyindole-3-acetic acid, tryptophol and others. The amounts detected in individual cells incubated in 5-hydroxytryptophan ranged from 10(-14) mol to 10(-16) mol, an order of magnitude higher than observed in untreated pinealocytes. |
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Authors:
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Christine Cecala; Stanislav S Rubakhin; Jennifer W Mitchell; Martha U Gillette; Jonathan V Sweedler |
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Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural Date: 2012-04-30 |
Journal Detail:
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Title: The Analyst Volume: 137 ISSN: 1364-5528 ISO Abbreviation: Analyst Publication Date: 2012 Jul |
Date Detail:
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Created Date: 2012-06-08 Completed Date: 2012-10-01 Revised Date: 2013-04-16 |
Medline Journal Info:
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Nlm Unique ID: 0372652 Medline TA: Analyst Country: England |
Other Details:
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Languages: eng Pagination: 2965-72 Citation Subset: IM |
Affiliation:
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Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Electrophoresis, Capillary / methods* Fluorescence Limit of Detection Pineal Gland / cytology Rats Single-Cell Analysis* |
| Grant Support | |
ID/Acronym/Agency:
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R01 DE018866/DE/NIDCR NIH HHS; R01 DE018866/DE/NIDCR NIH HHS; R01 HL086870/HL/NHLBI NIH HHS; R01 NS031609/NS/NINDS NIH HHS; R01 NS031609/NS/NINDS NIH HHS |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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