Abstract
Analyzing individual cells allows detecting a minor group of abnormal cells present in a large population of normal cells. This ability can be essential to understanding diseases, such as cancer and diabetes. Microchip electrophoresis (MCE) is the technique of choice for single-cell analysis. However, since the channels in microfluidic devices are very small, achieving the desired assay sensitivity on a microfluidic platform remains a challenge. Here, we describe an MCE method with highly sensitive chemiluminescence detection for simultaneous determination of multiple amino acids present in single cells.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
McClain MA et al (2003) Microfluidic devices for the high-throughput chemical analysis of cells. Anal Chem 75:5646–5655
Dufva M (2009) Microchips for cell-based assays. Methods Mol Biol 509:135–144
Kruth HS (1982) Flow cytometry: rapid biochemical analysis of single cells. Anal Biochem 125:225–242
Darzynkiewicz Z, Halicka HD, Zhao H (2010) Analysis of cellular DNA content by flow and laser scanning cytometry. Adv Exp Med Biol 676:137–47
Oates MD, Cooper BR, Jorgenson JW (1990) Quantitative amino acid analysis of individual snail neurons by open tubular liquid chromatography. Anal Chem 62:1573–1577
Troyer KP, Wightman RM (2002) Dopamine transport into a single cell in a picoliter vial. Anal Chem 74:5370–5375
Ishijima A, Yanagida T (2001) Single molecule nanobioscience. Trends Biochem Sci 26:438–444
Amantonico A et al (2010) Single-cell MALDI-MS as an analytical tool for studying intrapopulation metabolic heterogeneity of unicellular organisms. Anal Chem 82:7394–7400
Hogan BL, Yeung ES (1992) Determination of intracellular species at the level of a single erythrocyte via capillary electrophoresis with direct and indirect fluorescence detection. Anal Chem 64:2841–2845
Han FT, Lillard SJ (2002) Monitoring differential synthesis of RNA in individual cells by capillary electrophoresis. Anal Biochem 302:136–143
Hu S et al (2001) Capillary sodium dodecyl sulfate-DALT electrophoresis of proteins in a single human cancer cell. Electrophoresis 22:3677–3682
Zhang H, Jin WR (2004) Analysis of amino acids in individual human erythrocytes by capillary electrophoresis with electroporation for intracellular derivatization and laser-induced fluorescence detection. Electrophoresis 25:480–486
Fuller KM, Arriaga E A (2003) Analysis of individual acidic organelles by capillary electrophoresis with laser-induced fluorescence detection facilitated by the endocytosis of fluorescently labeled microspheres. Anal Chem 75:2123–2130
Zhi Q et al (2007) Coupling chemiluminescence with capillary electrophoresis to analyze single human red blood cells. Anal Chim Acta 583:217–222
Sims CE, Allbritton NL (2007) Analysis of single mammalian cells on-chip. Lab Chip 7:423–440
Huang WH et al (2008) Recent advances in single-cell analysis using capillary electrophoresis and microfluidic devices. J Chromatogr B 866:104–122
Wu H, Wheeler A, Zare RN (2004) Chemical cytometry on a picoliter-scale integrated microfluidic chip. Proc Natl Acad Sci USA 101:12809–12813
Ling YY, Yin XF, Fang ZL (2005) Simultaneous determination of glutathione and reactive oxygen species in individual cells by microchip electrophoresis. Electrophoresis 26:4759–4766
Klepárník K, Horký M (2003) Detection of DNA fragmentation in a single apoptotic cardiomyocyte by electrophoresis on a microfluidic device. Electrophoresis 24:3778–3783
Zhao S, Li X, Liu YM (2009) Integrated microfluidic system with chemiluminescence detection for single cell analysis after intracellular labeling. Anal Chem 81:3873–3878
Zhao S et al (2010) Determination of intracellular sulphydryl compounds by microchip electrophoresis with selective chemiluminescence detection. J Chromatogr A 1217:5732–5736
Jin W, Li X, Gao N (2003) Simultaneous determination of tryptophan and glutathione in individual rat hepatocytes by capillary zone electrophoresis with electrochemical detection at a carbon fiber bundle–Au/Hg dual electrode. Anal Chem 75:3859–3864
Kim M-S et al (2005) Fabrication of microchip electrophoresis devices and effects of channel surface properties on separation efficiency. Sensors and Actuators B 107:818–824.
Acknowledgments
Financial support from National Institutes of Health (SC1 GM089557) is gratefully acknowledged.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Liu, YM., Zhao, S. (2012). Quantification of Amino Acids in a Single Cell by Microchip Electrophoresis with Chemiluminescence Detection. In: Alterman, M., Hunziker, P. (eds) Amino Acid Analysis. Methods in Molecular Biology, vol 828. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-445-2_28
Download citation
DOI: https://doi.org/10.1007/978-1-61779-445-2_28
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-444-5
Online ISBN: 978-1-61779-445-2
eBook Packages: Springer Protocols