Document Detail

Capillary electrophoresis with three-color fluorescence detection for the analysis of glycosphingolipid metabolism.
MedLine Citation:
PMID:  23154386     Owner:  NLM     Status:  MEDLINE    
A capillary electrophoresis system with an ultrasensitive three-color laser-induced fluorescence detector was constructed for the simultaneous measurement of glycosphingolipids conjugated with a family of BODIPY fluorophores. The compounds were separated by capillary electrophoresis and detected by laser-induced fluorescence excited within a sheath-flow cuvette. Diode-pumped solid-state lasers operating at 473 nm and 532 nm, and a diode laser operating at 633 nm were used to excite glycosphingolipids tagged with BODIPY-FL, BODIPY-TMR, and BODIPY-650/665 fluorophores. Detection limits were 34 ± 1 molecules, 67 ± 7 molecules, and 36 ± 13 molecules of BODIPY-FL, BODIPY-TMR, and BODIPY-650/665 labeled glycosphingolipids. Separation efficiencies were typically one million theoretical plates. To test the ability of the system to analyze cellular contents in an in vitro biological model, differentiated PC12 cells were co-incubated with BODIPY-FL, BODIPY-TMR, and BODIPY-650/665 labeled lactosylceramide substrates. Cells were homogenized. The metabolic products originating from the glycosphingolipid substrates were simultaneously analyzed using the system.
Richard B Keithley; Alison S Rosenthal; David C Essaka; Hidenori Tanaka; Yayoi Yoshimura; Monica M Palcic; Ole Hindsgaul; Norman J Dovichi
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2012-11-16
Journal Detail:
Title:  The Analyst     Volume:  138     ISSN:  1364-5528     ISO Abbreviation:  Analyst     Publication Date:  2013 Jan 
Date Detail:
Created Date:  2012-11-28     Completed Date:  2014-01-08     Revised Date:  2014-01-16    
Medline Journal Info:
Nlm Unique ID:  0372652     Medline TA:  Analyst     Country:  England    
Other Details:
Languages:  eng     Pagination:  164-70     Citation Subset:  IM    
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MeSH Terms
Boron Compounds / chemistry
Electrophoresis, Capillary / methods*
Glycosphingolipids / chemistry,  metabolism*
PC12 Cells
Spectrometry, Fluorescence / instrumentation,  methods*
Grant Support
Reg. No./Substance:
0/4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; 0/Boron Compounds; 0/Glycosphingolipids

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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