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Finger-Actuated Microfluidic Concentration Gradient Generator Compatible with a Microplate
The generation of concentration gradients is an essential part of a wide range of laboratory settings. However, the task usually requires tedious and repetitive steps and it is difficult to generate concentration gradients at once. Here, we present a microfluidic device that easily generates a conce...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6471275/ https://www.ncbi.nlm.nih.gov/pubmed/30832320 http://dx.doi.org/10.3390/mi10030174 |
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author | Park, Juhwan Roh, Hyewon Park, Je-Kyun |
author_facet | Park, Juhwan Roh, Hyewon Park, Je-Kyun |
author_sort | Park, Juhwan |
collection | PubMed |
description | The generation of concentration gradients is an essential part of a wide range of laboratory settings. However, the task usually requires tedious and repetitive steps and it is difficult to generate concentration gradients at once. Here, we present a microfluidic device that easily generates a concentration gradient by means of push-button actuated pumping units. The device is designed to generate six concentrations with a linear gradient between two different sample solutions. The microfluidic concentration gradient generator we report here does not require external pumps because changes in the pressure of the fluidic channel induced by finger actuation generate a constant volume of fluid, and the design of the generator is compatible with the commonly used 96-well microplate. Generation of a concentration gradient by the finger-actuated microfluidic device was consistent with that of the manual pipetting method. In addition, the amount of fluid dispensed from each outlet was constant when the button was pressed, and the volume of fluid increased linearly with respect to the number of pushing times. Coefficient of variation (CV) was between 0.796% and 13.539%, and the error was between 0.111% and 19.147%. The design of the microfluidic network, as well as the amount of fluid dispensed from each outlet at a single finger actuation, can be adjusted to the user’s demand. To prove the applicability of the concentration gradient generator, an enzyme assay was performed using alkaline phosphatase (ALP) and para-nitrophenyl phosphate (pNPP). We generated a linear concentration gradient of the pNPP substrate, and the enzyme kinetics of ALP was studied by examining the initial reaction rate between ALP and pNPP. Then, a Hanes–Woolf plot of the various concentration of ALP was drawn and the V(max) and K(m) value were calculated. |
format | Online Article Text |
id | pubmed-6471275 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-64712752019-04-27 Finger-Actuated Microfluidic Concentration Gradient Generator Compatible with a Microplate Park, Juhwan Roh, Hyewon Park, Je-Kyun Micromachines (Basel) Article The generation of concentration gradients is an essential part of a wide range of laboratory settings. However, the task usually requires tedious and repetitive steps and it is difficult to generate concentration gradients at once. Here, we present a microfluidic device that easily generates a concentration gradient by means of push-button actuated pumping units. The device is designed to generate six concentrations with a linear gradient between two different sample solutions. The microfluidic concentration gradient generator we report here does not require external pumps because changes in the pressure of the fluidic channel induced by finger actuation generate a constant volume of fluid, and the design of the generator is compatible with the commonly used 96-well microplate. Generation of a concentration gradient by the finger-actuated microfluidic device was consistent with that of the manual pipetting method. In addition, the amount of fluid dispensed from each outlet was constant when the button was pressed, and the volume of fluid increased linearly with respect to the number of pushing times. Coefficient of variation (CV) was between 0.796% and 13.539%, and the error was between 0.111% and 19.147%. The design of the microfluidic network, as well as the amount of fluid dispensed from each outlet at a single finger actuation, can be adjusted to the user’s demand. To prove the applicability of the concentration gradient generator, an enzyme assay was performed using alkaline phosphatase (ALP) and para-nitrophenyl phosphate (pNPP). We generated a linear concentration gradient of the pNPP substrate, and the enzyme kinetics of ALP was studied by examining the initial reaction rate between ALP and pNPP. Then, a Hanes–Woolf plot of the various concentration of ALP was drawn and the V(max) and K(m) value were calculated. MDPI 2019-03-02 /pmc/articles/PMC6471275/ /pubmed/30832320 http://dx.doi.org/10.3390/mi10030174 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Park, Juhwan Roh, Hyewon Park, Je-Kyun Finger-Actuated Microfluidic Concentration Gradient Generator Compatible with a Microplate |
title | Finger-Actuated Microfluidic Concentration Gradient Generator Compatible with a Microplate |
title_full | Finger-Actuated Microfluidic Concentration Gradient Generator Compatible with a Microplate |
title_fullStr | Finger-Actuated Microfluidic Concentration Gradient Generator Compatible with a Microplate |
title_full_unstemmed | Finger-Actuated Microfluidic Concentration Gradient Generator Compatible with a Microplate |
title_short | Finger-Actuated Microfluidic Concentration Gradient Generator Compatible with a Microplate |
title_sort | finger-actuated microfluidic concentration gradient generator compatible with a microplate |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6471275/ https://www.ncbi.nlm.nih.gov/pubmed/30832320 http://dx.doi.org/10.3390/mi10030174 |
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