Analysis of single cells by capillary electrophoresis with on-column derivatization and laser-induced fluorescence detection.
On-column derivatization of single mammalian cells with capillary electrophoretic separation and laser-induced fluorescence detection is described. Individual cells are electrophorectically injected into the front end of the separation capillary, which is used as a chamber to lyse the cell and derivatize its contents for subsequent separation and detection. Reagents to lyse the cell and derivatize its contents are electrophoretically injected into the front end of the capillary (2.7 mm, 600-pL volume for a 17-microns-i.d. capillary) after the cell has been injected. Dopamine and five amino acids have been quantitatively determined in individual rat pheochromocytoma cells after on-column derivatization with naphthalene-2,3-dicarboxaldehyde and CN-. Average values of compounds determined in these cells range from 180 +/- 110 amol/cell for aspartic acid to 5.1 +/- 1.5 fmol/cell for taurine.
Gilman
S D
SD
Department of Chemistry, Pennsylvania State University, University Park 16802.
Ewing
A G
AG
D000588:Amines / Q000032:analysis; D000595:Amino Acid Sequence; D000818:Animals; D002477:Cells / Q000737:chemistry*; D004298:Dopamine / Q000032:analysis; D004586:Electrophoresis / Q000379:methods*; D007834:Lasers; D008969:Molecular Sequence Data; D016716:PC12 Cells; D051381:Rats; D013050:Spectrometry, Fluorescence / Q000379:methods*
D000588:Amines; D004298:Dopamine