Abstract
We present a method suitable for quantitative analysis of intracellular proteins, metabolites and secondary messengers of single bacterial cells. The method integrates the concept of immunoassays on a microfluidic device that facilitates single cell trapping and isolating in a small volume of a few tens of picoliters. Combination of the benefits of microfluidic systems for single cell analysis with the high analytical selectivity and sensitivity of immunoassays enables the detection of even low abundant intracellular analytes which occur only at a few hundred copies per bacterium.
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
References
Kaern M, Elston TC, Blake WJ et al (2005) Stochasticity in gene expression: from theories to phenotypes. Nat Rev Genet 6:451–464
Ito Y, Toyota H, Kaneko K et al (2009) How selection affects phenotypic fluctuation. Mol Syst Biol 5:1–7
Munsky B, Neuert G, Van Oudenaarden A (2012) Using gene expression noise to understand gene regulation. Science 336:183–187
Viney M, Reece SE (2013) Adaptive noise. Proc R Soc B 280:1–9
Hunt BG, Ometto L, Keller L et al (2013) Evolution at two levels in fire ants: the relationship between patterns of gene expression and protein sequence evolution. Mol Biol Evol 30:263–271
Taniguchi Y, Choi PJ, Li GW, Chen H et al (2010) Quantifying E. coli proteome and transcriptome with single-molecule sensitivity in single cells. Science 329:533–538
Mazumder A, Tummler K, Bathe M et al (2013) Single-cell analysis of ribonucleotide reductase transcriptional and translational response to DNA damage. Mol Cell Biol 33:635–642
Dittrich PS, Manz A (2006) Lab-on-a-chip: microfluidics in drug discovery. Nat Rev Drug Discov 5:210–218
KlepárnÃk K, Foret F (2013) Recent advances in the development of single cell analysis—a review. Anal Chim Acta 800:12–21
Kovarik ML, Gach PC, Orno DM et al (2012) Micro total analysis systems for cell biology and biochemical assays. Anal Chem 84:516–540
Zhang Y, Ozdemir P (2009) Microfluidic DNA amplification—a review. Anal Chim Acta 638:115–125
Lounsbury JA, Karlsson A, Miranian DC et al (2013) From sample to PCR product in under 45 minutes: a polymeric integrated microdevice for clinical and forensic DNA analysis. Lab Chip 13:1384–1393
Chang CM, Chang WH, Wang CH et al (2013) Nucleic acid amplification using microfluidic systems. Lab Chip 13:1225–1242
Wu M, Singh AK (2012) Single-cell protein analysis. Curr Opin Biotechnol 23:83–88
Bendall SC, Simonds EF, Qiu P et al (2011) Single-cell mass cytometry of differential immune and drug responses across a human hematopoietic continuum. Science 332:687–696
Eyer K, Stratz S, Dittrich PS et al (2013) Implementing enzyme-linked immunosorbent assays on a microfluidic chip to quantify intracellular molecules in single cells. Anal Chem 85:3280–3287
Eyer K, Kuhn P, Dittrich PS et al (2012) A microchamber array for single cell isolation and analysis of intracellular biomolecules. Lab Chip 12:765–772
Chen Y, Zhang B, Feng H et al (2012) An automated microfluidic device for assessment of mammalian cell genetic stability. Lab Chip 12:3930–3935
Leung K, Zahn H, Leaver T et al (2012) A programmable droplet-based microfluidic device applied to multiparameter analysis of single microbes and microbial communities. Proc Natl Acad Sci U S A 109:7665–7670
Kim M, Isenberg BC, Sutin J et al (2011) Programmed trapping of individual bacteria using micrometer-size sieves. Lab Chip 11:1089–1095
He M, Edgar JS, Jeffries GD et al (2005) Selective encapsulation of single cells and subcellular organelles into picoliter- and femtoliter-volume droplets. Anal Chem 77:1539–1544
Stratz S, Eyer K, Kurth F, Dittrich PS (2014) On-chip enzyme quantification of single Escherichia coli bacteria by immunoassay-based analysis. Anal Chem 86: 12375–12381
Berg JM, Tymoczko JL, Stryer L (2002) The Michaelis-Menten model accounts for the kinetic properties of many enzymes. In: Freeman WH (ed) Biochemistry, 5th edn. Freeman W. H. and Company, New York
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2015 Springer Science+Business Media New York
About this protocol
Cite this protocol
Stratz, S., Dittrich, P.S. (2015). A Microfluidic Device for Immunoassay-Based Protein Analysis of Single E. coli Bacteria. In: Singh, A., Chandrasekaran, A. (eds) Single Cell Protein Analysis. Methods in Molecular Biology, vol 1346. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2987-0_2
Download citation
DOI: https://doi.org/10.1007/978-1-4939-2987-0_2
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2986-3
Online ISBN: 978-1-4939-2987-0
eBook Packages: Springer Protocols