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Article

Characterization and Optimization of Isotachophoresis Parameters for Pacific Blue Succinimidyl Ester Dye on a PDMS Microfluidic Chip

1
Department of Mechanical Engineering, Texas Tech University, Lubbock, TX 79409, USA
2
Department of Mechanical Engineering, University of Utah, Salt Lake City, UT 84112, USA
3
KIST-EUROPE, 66123 Saarbrucken, Germany
*
Author to whom correspondence should be addressed.
Micromachines 2020, 11(11), 951; https://doi.org/10.3390/mi11110951
Received: 16 September 2020 / Revised: 20 October 2020 / Accepted: 20 October 2020 / Published: 22 October 2020
(This article belongs to the Special Issue Electrokinetics in Micro-/nanofluidic Devices)
Isotachophoresis (ITP) for Pacific Blue (PB) dye using a polydimethylsiloxane (PDMS) microfluidic chip is developed and characterized by determining the types and concentrations of electrolytes, the ITP duration, and the electric field density. Among candidate buffers for the trailing electrolyte (TE) and leading electrolyte (LE), 40 mM borate buffer (pH 9) and 200 mM trisaminomethane hydrochloride (Tris-HCl) (pH 8) were selected to obtain the maximum preconcentration and resolution of the PB bands, respectively. With the selected TE and LE buffers, further optimization was performed to determine the electric field (EF) density and the ITP duration. These ITP parameters showed a 20–170,000 preconcentration ratio from initial PB concentrations of 10 nM–100 fM. Further demonstration was implemented to preconcentrate PB-conjugated lactate dehydrogenase (LDH) using the PDMS microfluidic chip. By utilizing the quenching nature of PB-LDH conjugation, we were able to identify concentrations of LDH as low as 10 ng/mL. This simple PDMS microfluidic chip-based ITP for PB preconcentration enables highly sensitive biological and chemical analyses by coupling with various downstream detection systems. View Full-Text
Keywords: isotachophoresis; PDMS (polydimethylsiloxane) microfluidic chip; pacific blue succinimidyl ester; preconcentration; lactate dehydrogenase isotachophoresis; PDMS (polydimethylsiloxane) microfluidic chip; pacific blue succinimidyl ester; preconcentration; lactate dehydrogenase
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MDPI and ACS Style

Somaweera, H.; Estlack, Z.; Devadhasan, J.P.; Kim, J.; Kim, J. Characterization and Optimization of Isotachophoresis Parameters for Pacific Blue Succinimidyl Ester Dye on a PDMS Microfluidic Chip. Micromachines 2020, 11, 951. https://doi.org/10.3390/mi11110951

AMA Style

Somaweera H, Estlack Z, Devadhasan JP, Kim J, Kim J. Characterization and Optimization of Isotachophoresis Parameters for Pacific Blue Succinimidyl Ester Dye on a PDMS Microfluidic Chip. Micromachines. 2020; 11(11):951. https://doi.org/10.3390/mi11110951

Chicago/Turabian Style

Somaweera, Himali, Zachary Estlack, Jasmine Pramila Devadhasan, Jungtae Kim, and Jungkyu Kim. 2020. "Characterization and Optimization of Isotachophoresis Parameters for Pacific Blue Succinimidyl Ester Dye on a PDMS Microfluidic Chip" Micromachines 11, no. 11: 951. https://doi.org/10.3390/mi11110951

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