Single-molecule enzymatic analysis in a droplet-based microfluidic system

Rerngchai Arayanarakool; Lingling Shui; S.W.M Kengen; Albert van den Berg; Jan C.T. Eijkel

Single-molecule enzymatic analysis in a droplet-based microfluidic system

Rerngchai Arayanarakool, Lingling Shui, S.W.M Kengen, Albert van den Berg, Jan C.T. Eijkel

Research output: Chapter in Book/Report/Conference proceeding › Conference contribution › Academic › peer-review

Abstract

The kinetic activity of individual enzyme molecules was determined in aqueous droplets generated in a nano- and microfluidic device. To avoid high background noise, the enzyme and substrate solution was confined into femtolitre carriers to achieve single-molecule encapsulation. The tiny droplets (f~2.5-3 μm) generated from this fluidic system were highly monodisperse, beneficial for an analysis of single enzyme activity. Single-enzyme kinetics has previously been demonstrated in the microfluidic format in PDMS containers [1], surface-immobilized droplets [2], or liposomes [3]. Single- enzyme analysis in the droplet-based microfluidics was reported before [4] by encapsulating highly-diluted enzyme solution (110 fM) into large droplets (f~40 μm). But to our knowledge this is the first demonstration of the direct method of single enzyme encapsulation and analysis at high enzyme concentration in tiny droplets in a microfluidic system.

Original language	Undefined
Title of host publication	The 16th International Conference on Miniaturized Systems for Chemistry and Life Sciences
Editors	T. Fujii, A. Hibara, S. Takeuchi, T. Fukuba
Publisher	The Chemical and Biological Microsystems Society
Pages	1492-1494
Number of pages	3
ISBN (Print)	1556-5904
Publication status	Published - 29 Oct 2012
Event	16th International Conference on Miniaturized Systems for Chemistry and Life Sciences, µTAS 2012 - Okinawa, Japan Duration: 28 Oct 2012 → 1 Nov 2012 Conference number: 16

Publication series

Name	microTAS
Publisher	Chemical and Biological Microsystems Society
ISSN (Print)	1556-5904

Conference

Conference	16th International Conference on Miniaturized Systems for Chemistry and Life Sciences, µTAS 2012
Abbreviated title	MicroTAS
Country/Territory	Japan
City	Okinawa
Period	28/10/12 → 1/11/12

Keywords

IR-83542
METIS-296177
EWI-22719

Access to Document

http://eprints.eemcs.utwente.nl/secure2/22719/01/W.1.18.pdf

Cite this

Arayanarakool, R., Shui, L., Kengen, S. W. M., van den Berg, A., & Eijkel, J. C. T. (2012). Single-molecule enzymatic analysis in a droplet-based microfluidic system. In T. Fujii, A. Hibara, S. Takeuchi, & T. Fukuba (Eds.), The 16th International Conference on Miniaturized Systems for Chemistry and Life Sciences (pp. 1492-1494). (microTAS). The Chemical and Biological Microsystems Society. http://eprints.eemcs.utwente.nl/secure2/22719/01/W.1.18.pdf

Arayanarakool, Rerngchai ; Shui, Lingling ; Kengen, S.W.M ; van den Berg, Albert ; Eijkel, Jan C.T. / Single-molecule enzymatic analysis in a droplet-based microfluidic system. The 16th International Conference on Miniaturized Systems for Chemistry and Life Sciences. editor / T. Fujii ; A. Hibara ; S. Takeuchi ; T. Fukuba. The Chemical and Biological Microsystems Society, 2012. pp. 1492-1494 (microTAS).

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title = "Single-molecule enzymatic analysis in a droplet-based microfluidic system",

abstract = "The kinetic activity of individual enzyme molecules was determined in aqueous droplets generated in a nano- and microfluidic device. To avoid high background noise, the enzyme and substrate solution was confined into femtolitre carriers to achieve single-molecule encapsulation. The tiny droplets (f~2.5-3 μm) generated from this fluidic system were highly monodisperse, beneficial for an analysis of single enzyme activity. Single-enzyme kinetics has previously been demonstrated in the microfluidic format in PDMS containers [1], surface-immobilized droplets [2], or liposomes [3]. Single- enzyme analysis in the droplet-based microfluidics was reported before [4] by encapsulating highly-diluted enzyme solution (110 fM) into large droplets (f~40 μm). But to our knowledge this is the first demonstration of the direct method of single enzyme encapsulation and analysis at high enzyme concentration in tiny droplets in a microfluidic system.",

keywords = "IR-83542, METIS-296177, EWI-22719",

author = "Rerngchai Arayanarakool and Lingling Shui and S.W.M Kengen and {van den Berg}, Albert and Eijkel, {Jan C.T.}",

year = "2012",

month = oct,

day = "29",

language = "Undefined",

isbn = "1556-5904",

series = "microTAS",

publisher = "The Chemical and Biological Microsystems Society",

pages = "1492--1494",

editor = "T. Fujii and A. Hibara and S. Takeuchi and T. Fukuba",

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Arayanarakool, R, Shui, L, Kengen, SWM, van den Berg, A & Eijkel, JCT 2012, Single-molecule enzymatic analysis in a droplet-based microfluidic system. in T Fujii, A Hibara, S Takeuchi & T Fukuba (eds), The 16th International Conference on Miniaturized Systems for Chemistry and Life Sciences. microTAS, The Chemical and Biological Microsystems Society, pp. 1492-1494, 16th International Conference on Miniaturized Systems for Chemistry and Life Sciences, µTAS 2012, Okinawa, Japan, 28/10/12. <http://eprints.eemcs.utwente.nl/secure2/22719/01/W.1.18.pdf>

Single-molecule enzymatic analysis in a droplet-based microfluidic system. / Arayanarakool, Rerngchai; Shui, Lingling; Kengen, S.W.M; van den Berg, Albert ; Eijkel, Jan C.T.

The 16th International Conference on Miniaturized Systems for Chemistry and Life Sciences. ed. / T. Fujii; A. Hibara; S. Takeuchi; T. Fukuba. The Chemical and Biological Microsystems Society, 2012. p. 1492-1494 (microTAS).

Research output: Chapter in Book/Report/Conference proceeding › Conference contribution › Academic › peer-review

TY - GEN

T1 - Single-molecule enzymatic analysis in a droplet-based microfluidic system

AU - Arayanarakool, Rerngchai

AU - Shui, Lingling

AU - Kengen, S.W.M

AU - van den Berg, Albert

AU - Eijkel, Jan C.T.

N1 - Conference code: 16

PY - 2012/10/29

Y1 - 2012/10/29

N2 - The kinetic activity of individual enzyme molecules was determined in aqueous droplets generated in a nano- and microfluidic device. To avoid high background noise, the enzyme and substrate solution was confined into femtolitre carriers to achieve single-molecule encapsulation. The tiny droplets (f~2.5-3 μm) generated from this fluidic system were highly monodisperse, beneficial for an analysis of single enzyme activity. Single-enzyme kinetics has previously been demonstrated in the microfluidic format in PDMS containers [1], surface-immobilized droplets [2], or liposomes [3]. Single- enzyme analysis in the droplet-based microfluidics was reported before [4] by encapsulating highly-diluted enzyme solution (110 fM) into large droplets (f~40 μm). But to our knowledge this is the first demonstration of the direct method of single enzyme encapsulation and analysis at high enzyme concentration in tiny droplets in a microfluidic system.

AB - The kinetic activity of individual enzyme molecules was determined in aqueous droplets generated in a nano- and microfluidic device. To avoid high background noise, the enzyme and substrate solution was confined into femtolitre carriers to achieve single-molecule encapsulation. The tiny droplets (f~2.5-3 μm) generated from this fluidic system were highly monodisperse, beneficial for an analysis of single enzyme activity. Single-enzyme kinetics has previously been demonstrated in the microfluidic format in PDMS containers [1], surface-immobilized droplets [2], or liposomes [3]. Single- enzyme analysis in the droplet-based microfluidics was reported before [4] by encapsulating highly-diluted enzyme solution (110 fM) into large droplets (f~40 μm). But to our knowledge this is the first demonstration of the direct method of single enzyme encapsulation and analysis at high enzyme concentration in tiny droplets in a microfluidic system.

KW - IR-83542

KW - METIS-296177

KW - EWI-22719

M3 - Conference contribution

SN - 1556-5904

T3 - microTAS

SP - 1492

EP - 1494

BT - The 16th International Conference on Miniaturized Systems for Chemistry and Life Sciences

A2 - Fujii, T.

A2 - Hibara, A.

A2 - Takeuchi, S.

A2 - Fukuba, T.

PB - The Chemical and Biological Microsystems Society

Y2 - 28 October 2012 through 1 November 2012

ER -