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The Key Laboratory of Biomedical Photonics of MOE, Hubei Bioinformatics & Molecular Imaging Key Laboratory, Department of Systems Biology,
College of Life Science & Technology, Huazhong University of Science and Technology, Wuhan 430074, China
b Graduate School of Material Science, University of Hyogo, Hyogo 678-1297, Japan
Received 28 August 2006; received in revised form 18 October 2006; accepted 29 October 2006
Available online 7 November 2006
Abstract
Electrokinetic gating, functioning as a micro-valve, has been widely employed in microfluidic chips for sample injection and flow switch.
Investigating its valving performance is fundamentally vital for microfluidics and microfluidics-based chemical analysis. In this paper, electrokinetic
gating valve in microchannels was evaluated using optical imaging technique. Microflow profiles at channels junction were examined, revealing
that molecular diffusion played a significant role in the valving disable; which could cause analyte leakage in sample injection. Due to diffusion,
the analyte crossed the interface of the analyte flow and gating flow, and then formed a cometic tail-like diffusion area at channels junction. From
theoretical calculation and some experimental evidences, the size of the area was related to the diffusion coefficient and the velocity of analytes.
Additionally, molecular diffusion was also believed to be another reason of sampling bias in gated injection.
2006 Elsevier B.V. All rights reserved.
Keywords: Electrokinetic gating; Microfluidic chip; Valving; Leakage
1. Introduction
Over the past decade, analysis on a microfluidic device has
attracted great attention, which leads to currently fast-growing
community of miniaturized total analysis system (TAS) [14].
Due to high-throughput potential, capabilities of miniaturization, integration and automation that represent the development
direction of next-generation instrumentation, TAS has been
recognized as a powerful tool in far-reaching application fields
[47], such as chemistry, biology, medicine, environment, and
pharmaceutics as well.
In TAS, there are several key modules or elements that functionally achieve microfluidic operations including actuating,
pumping, valving or switching, mixing, sensing and separating etc., which form the solid basis of so-called lab-on-a-chip.
Thus, to give a deep insight into these elements is fundamentally vital, as it will greatly help us to understand the mechanism
of microfluidics and further to construct the state-of-the-art
Corresponding author. Tel.: +86 27 8779 2170; fax: +86 27 8779 2203.
E-mail address: bfliu@mail.hust.edu.cn (B.-F. Liu).
0003-2670/$ see front matter 2006 Elsevier B.V. All rights reserved.
doi:10.1016/j.aca.2006.10.046
130
convenience for flow switch between channels in multidimensional separation on a microchip [31]. However, it has seldom
been experimentally investigated in detail, for example, sampling bias or leakage. Ermakov et al. [30] demonstrated a
computational simulation of gating valving. Processes of sample
loading and dispensing were theoretically calculated. Recently
Slentz et al. [29] discussed sample bias at microchannel junctions in gating injection. They found a new type of sampling bias
beyond electrokinetic discrimination, called transradial electrokinetic selection (TREKS). Based on this phenomenon, the
authors further suggested a diffusion injection method to correct such a sampling bias by TREKS. But it was still unclear the
underlying mechanism TREKS occurred.
Leakage is a significant criteria to evaluate the quality of
a valve. Zero leakage is required for a reliable valve. Since
electrokinetic gating functions based on microfluidic interaction, not actual physic barrier, leakage shows different features
from mechanical or magnetic valves simulated by Ermakov et
al. [30]. To reveal the underlying mechanism, it is fundamentally
important to examine the transportation behaviors of molecules
at microchannels junction.
For this consideration, electrokinetic gating valve in
microfluidic channels was characterized in this study. The
microflow profiles at channels junction were monitored and analyzed using fluorescence imaging technique. It was suggested
that molecule diffusion was the key factor of the leakage of
electrokinetic valve, and also cause molecular discrimination in
gated injection.
Japan). The microchip was fabricated using standard photolithograph, wet chemical etching, and bonding techniques as
previously described [32]. The channel was 80 m in width on
the top and 40 m in depth. For new microchip, channels were
washed sequentially with methanol, water, 1 M NaOH, 0.1 M
HCl, 0.1 M NaOH, water and running buffer at room temperature. Between analysis, channels were rinsed in an order of 0.1 M
NaOH, water and running buffer to ensure the reproducibility.
2.4. Electrokinetic gating
In gated injection or flow switch in two-dimensional separation on a microchip, a microflow initially carried the analytes
from Channel S to Channel SW. To prevent the analytes from
leaking into Channel BW, another microflow from Channel B
was introduced into Channel SW and Channel BW simultaneously, as shown in Fig. 1A. When the microflow from Channel B
was stopped, the analytes were then injected from Channel S into
Channel BW (Fig. 1B). Afterwards, the microflow from Channel B was recovered (Fig. 1C). The analytes were gated again.
Microfluidic program above could be conveniently achieved by
2. Experimental
2.1. Materials
Fluorescein isothiocyanate (FITC) was obtained from
SigmaAldrich (MO, US). Rhodamine series dyes including
rhodamine 6G, rhodamine B and sulforhodamine were purchased from TCI Chemicals (Tokyo, Japan). All other reagents
were of analytical grade. Water purified by the Milli-Q system
(Millipore, US) was used for the preparations of all solutions.
2.2. Instrument
Visualization and imaging of flow profiles at the microchannel junction were carried out on an inverted fluorescence
microscope (Eclipse TS 100-F, Nikon, Tokyo, Japan) equipped
with a 3CCD digital camera (HV-D28S, Hitachi Kokusai Electric Inc., Tokyo, Japan). Acquired fluorescent images were
further analyzed using Scion Image software package (NIH,
US). Power supply (Matsusada Precision Devices, Japan) for
flow pumping was computer-controlled with terminal emulation
software KTX (freeware) connected to a D/A converter (Nippon
Filcon, Inagi, Japan) via an RS-232C serial interface.
2.3. Experimental procedures
The glass microchip with a channel pattern of a cross was
kindly obtained from Kitamori group (University of Tokyo,
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Fig. 2. Analyte leakage at g = 1.2. FITC was employed as the analyte dissolved
in buffer of 20 mM PBS (pH 8.5), and rhodamine B was added into buffer
solution for fluorescent visualization. (A) Micrograph using filters for FITC.
(B) Micrograph using filters for rhodamine B.
Fig. 3. Electropherogram of four fluorescent species using capillary electrophoresis on a microchip. Buffer, 20 mM PBS (pH 8.5). Electric field strength
for separation, 330 V cm1 . Gated injection, 330 V cm1 for 0.2 s. Peak identity,
1, rhodamine 6G; 2, rhodamine B; 3, sulforhodamine; 4, FITC.
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Fig. 6. Influence of electric field strength on analyte profiling at the channels junction. (A) 500 V cm1 ; (B) 400 V cm1 ; (C) 300 V cm1 ; (D) 200 V cm1 ; (E)
100 V cm1 ; (F) schematic description of analyte profiling. Curves identity, 0, interface of analyte flow and gating flow; 15, boundaries the analyte could diffused
to in different electric field strengths at the channels junction (AE), respectively.
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(1)
(2)
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the analyte flow was electrokinetic gated by the gating flow, the
analyte would diffuse through the interface of the two flows, and
formed a cometic tail-like diffusion area at the channels junction. Because the size of the diffusion area was dependent of
molecular diffusion coefficient and electrophoretic mobility, it
was not surprised why sampling bias could happen by TREKS
[29].
4. Conclusion
Fig. 8. Gaussian fit of data points from intensity profiling in Line 1/Fig. 4C.
Data was symmetrically extended. Hollow and solid circles symbolized original
data and extended data, respectively.
(3)
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