Document Detail


A hyphenated optical trap capillary electrophoresis laser induced native fluorescence system for single-cell chemical analysis.
MedLine Citation:
PMID:  22543409     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Christine Cecala; Stanislav S Rubakhin; Jennifer W Mitchell; Martha U Gillette; Jonathan V Sweedler
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2012-04-30
Journal Detail:
Title:  The Analyst     Volume:  137     ISSN:  1364-5528     ISO Abbreviation:  Analyst     Publication Date:  2012 Jul 
Date Detail:
Created Date:  2012-06-08     Completed Date:  2012-10-01     Revised Date:  2013-07-12    
Medline Journal Info:
Nlm Unique ID:  0372652     Medline TA:  Analyst     Country:  England    
Other Details:
Languages:  eng     Pagination:  2965-72     Citation Subset:  IM    
Affiliation:
Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Electrophoresis, Capillary / methods*
Fluorescence
Limit of Detection
Pineal Gland / cytology
Rats
Single-Cell Analysis*
Grant Support
ID/Acronym/Agency:
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
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