Document Detail


Capillary electrophoresis with three-color fluorescence detection for the analysis of glycosphingolipid metabolism.
MedLine Citation:
PMID:  23154386     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Richard B Keithley; Alison S Rosenthal; David C Essaka; Hidenori Tanaka; Yayoi Yoshimura; Monica M Palcic; Ole Hindsgaul; Norman J Dovichi
Related Documents :
18351296 - A general approach to the apparent permeability index.
20389606 - Intermediate high index layer for laser mode tuning in organic semiconductor lasers.
20198086 - Retroreflection from spherical glass beads in highway pavement markings. 2: diffuse ref...
12366286 - Calculations and measurements of wire and/or split-ring negative index media.
18950136 - Importance of van der waals interactions in qm/mm simulations.
18354586 - Thermal emission from dielectric beam splitters in michelson interferometers: a schemat...
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    
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Animals
Boron Compounds / chemistry
Color
Electrophoresis, Capillary / methods*
Glycosphingolipids / chemistry,  metabolism*
Lasers
PC12 Cells
Rats
Spectrometry, Fluorescence / instrumentation,  methods*
Grant Support
ID/Acronym/Agency:
R01 NS061767/NS/NINDS NIH HHS; R01NS061767/NS/NINDS NIH HHS
Chemical
Reg. No./Substance:
0/4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; 0/Boron Compounds; 0/Glycosphingolipids
Comments/Corrections

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


Previous Document:  Ascorbic acid improves the developmental competence of porcine oocytes after parthenogenetic activat...
Next Document:  The Ca(2+)/Mn(2+) ion-pump PMR1 links elevation of cytosolic Ca(2+) levels to ?-synuclein toxicity i...