Single neuron analysis by capillary electrophoresis with fluorescence spectroscopy.
A technique to identify and quantitate simultaneously more than 30 compounds in individual neurons is described. The method uses nanoliter volume sampling, capillary electrophoresis separation, and wavelength-resolved native fluorescence detection. Limits of detection (LODs) range from the low attomole to the femtomole range, with 5-hydroxytryptamine (or serotonin [5-HT]) LODs being approximately 20 attomoles. Although the cellular sample matrix is chemically complex, the combination of electrophoretic migration time and fluorescence spectral information allows positive identification of aromatic monoamines, aromatic amino acids and peptides containing them, flavins, adenosine- and guanosine-nucleotide analogs, and other fluorescent compounds. Individual identified neurons from Aplysia californica and Pleurobranchaea californica are used to demonstrate the applicability and figures of merit of this technique.
Fuller
R R
RR
Department of Chemistry and the Beckman Institute, University of Illinois, Urbana 61801, USA.
Moroz
L L
LL
Gillette
R
R
Sweedler
J V
JV
D000818:Animals; D001048:Aplysia / Q000502:physiology*; D003627:Data Interpretation, Statistical; D019075:Electrophoresis, Capillary / Q000379:methods* / Q000592:standards; D009474:Neurons / Q000737:chemistry / Q000502:physiology*; D012015:Reference Standards; D013050:Spectrometry, Fluorescence / Q000379:methods* / Q000592:standards