Capillary electrophoresis with electrospray ionization mass spectrometric detection for single-cell metabolomics.
A method that enables metabolomic profiling of single cells and subcellular structures is described using capillary electrophoresis coupled to electrospray ionization time-of-flight mass spectrometry. A nebulizer-free coaxial sheath-flow interface completes the circuit and provides a stable electrospray, yielding a signal with a relative standard deviation of under 5% for the total ion electropherogram. Detection limits are in the low nanomolar range (i.e., <50 nM (<300 amol)) for a number of cell-to-cell signaling molecules, including acetylcholine (ACh), histamine, dopamine, and serotonin. The instrument also yields high-efficiency separations, e.g., approximately 600,000 for eluting ACh bands. The utility of this setup for single-cell metabolomic profiling is demonstrated with identified neurons from Aplysia californica--the R2 neuron and metacerebral cell (MCC). Single-cell electropherograms are reproducible, with a large number of metabolites detected; more than 100 compounds yield signals of over 10(4) counts from the injection of only 0.1% of the total content from a single MCC. Expected neurotransmitters are detected within the cells (ACh in R2 and serotonin in MCC), as are compounds that have molecular masses consistent with all of the naturally occurring amino acids (except cysteine). Tandem MS using a quadrupole time-of-flight tandem mass spectrometer distinguishes ACh from isobaric compounds in the R2 neuron and demonstrates the ability of this method to characterize and identify metabolites present within single cells.
Lapainis
Theodore
T
Department of Chemistry and the Beckman Institute, University of Illinois, Urbana, Illinois 61801, USA.
Rubakhin
Stanislav S
SS
Sweedler
Jonathan V
JV
D000818:Animals; D001048:Aplysia / Q000166:cytology / Q000378:metabolism; D019075:Electrophoresis, Capillary; D042541:Intracellular Space / Q000378:metabolism; D055432:Metabolomics / Q000379:methods*; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry