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Sensors and Actuators B 136 (2009) 265274

Contents lists available at ScienceDirect

Sensors and Actuators B: Chemical


journal homepage: www.elsevier.com/locate/snb

Versatile programmable eight-path-electrode power supply for automatic


manipulating microuids of a microuidic chip
Qingling Li, Hui Zhang, Yan Wang, Bo Tang , Xin Liu, Xiaocong Gong
College of Chemistry, Chemical Engineering and Materials Science, Key Laboratory of Molecular and Nano Probes, Ministry of Education, Shandong Normal University,
Jinan 250014, China

a r t i c l e

i n f o

Article history:
Received 11 April 2008
Received in revised form 14 August 2008
Accepted 21 October 2008
Available online 17 November 2008
Keywords:
Microuids
Microuid manipulation
Programmable control
Multi-path power supply
Microuidic chip

a b s t r a c t
A versatile programmable eight-path-electrode power supply (PEPS) system for manipulating microuids
of a complex microuidic chip has been developed. The PEPS system consisted of a single chip microprocessor as a central control unit and a personal computer (PC) as an upper computer, and the program
could be operated under Windows98/2000/XP. The voltage output of each electrode was in the range
of 0 to +8000 V (0.1% precision) while the current output was in the range of 0 to +999 A. The voltage
of eight electrodes could be operated either independently or synchronously by random combination of
any electrodes through switching. The voltage output modes were switch-off/oating, switch-on and
grounded and fast switched at ms-level between these modes, and run time (0.1 s precision) of these
modes could be controlled as desired. The PEPS system was conveniently for controlling ow rate and
direction of electroosmotic ow (EOF) in a chip network. Six electrodes were chosen to control the repeated
injection and separation of 1.0 105 M uorescein isothiocyanate (FITC) in a six-reservoir glass-based
chip. The relative standard deviation (R.S.D., n = 4, S/N = 10) of the repeated operation was 0.9% for the
reservation time (tR ) and 2.3% for the peak height, respectively.
2008 Published by Elsevier B.V.

1. Introduction
Microuidic chip or micrototal analysis system (TAS) provides
a new technology platform for the research in chemistry, biology, microengineering and other related microsystem elds [1,2].
Microuidic chip devices offer many attractive benets such as
reducing device size, reducing sample requirements and reagent
volumes which can reduce overall cost. Combining different functions on a single microchip is an important step toward maintaining
a completely closed system that can be fully automated, reduce
contamination and eliminate human intervention and error [3].
However, in order to realize all above the benets, it is imperative to solve innovatively the problem of automatic control of
the microuids. The microuid control was dependent on two
categories of micropump techniques, mechanical displacement
micropump technique and non-mechanical micropump technique
[46]. Mechanical micropumps [4] such as syringe pumps [7], pneumatic pumps [8,9] and piezoelectric pumps [10]were dened as
those that exert oscillatory or rotational pressure forces on the
working uid through a moving soliduid (vibrating diaphragm,
peristaltic, rotary pumps), or uiduid boundary (ferrouid, phase

Corresponding author. Tel.: +86 531 86180010; fax: +86 531 86180017.
E-mail address: tangb@sdnu.edu.cn (B. Tang).
0925-4005/$ see front matter 2008 Published by Elsevier B.V.
doi:10.1016/j.snb.2008.10.066

change, gas permeation pumps). Non-mechanical micropump, i.e.


continuous ow kinetic micropump [5,6], such as magnetic pumps
[11] and electroosmotic pumps [46,1214]were dened as those
that provide a direct energy transfer to pumping power and
generate constant/steady ows (electrokinetic, magnetokinetic,
electrowetting, etc.). Valveless continuous dynamic ow micropump technique has now become the rapidly developed research
area for its simple structure and powerful function [6].
One of the development trends of TAS was the chip with more
complicated channel network and more functional units. More
concern to the chip were the uids delivery and the ow rate
in the range of 100 pL/min to several L/min (without regard to
the pressure) and the effective control of the magnitude, compose
and direction of the uids in the microchannel network [15,16].
The electroosmotic pumps (EOP) based on the electroosmotic ow
(EOF) principle are still the most used techniques for microscale
microuid delivery in the chip because of its capability to perform automatic microuidic manipulation with stable ow rate
and appropriate ow quantity [6,16]. The EOPs are convenient
to control the uid ow and direction by regulating the voltage
applied to the microchannel network and electric eld direction.
For example, in chip-based capillary electrophoresis (CE) process,
the sampling volume could be controlled by regulating the voltage of the sampling channel, forming a steady sample zone, and
then switching the voltage applied to the separation channel for

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Q. Li et al. / Sensors and Actuators B 136 (2009) 265274

separation. The switch of microuids direction was convenient


since no valve was needed in the whole operation process. Therefore, the programmable multi-channel voltage sequencer is urgent
for complex microuids automatic manipulation. Presently, the
most reported power supplies were 4-path electrode which could
be used for cross or double T chip with four reservoirs. Garcia et al. [17] reported a three-path-electrode sequencer which
could complete the CE injection and separation on a double-T
chip. Fletcher et al. [18] described a kind of four-path-electrode
sequencer which could complete the injection under electrokinetic
force on a double-T chip. Wu et al. [19] reported an electricaldriven instrument with a single power supply connected to a
controlled eight-position relay box. The voltage obtained by the
sub-voltage from attenuation and channel electric resistance could
not be automatically adjusted. The disadvantage of the reported
voltage sequencer used for chip was that the limited-path voltage output which could not be used for multi-channel microuidic
chip with ve to eight reservoirs. Obviously, a programmable multipath power supply was often required for electrokinetic driving in
microuid multichannel network of these chips. In this work, a programmable eight-path-electrode power supply (PEPS) system was
developed for effectively controlling the ow rate and direction of
microuids of ve to eight reservoirs microuidic chip in the multichannel network [20]. The voltages of eight electrodes, which could
be operated either independently or synchronously by random
combination any electrodes under the control of program, could
conveniently and automatically control ow rate and direction of
microscale uids. Compared with commercial voltage sequencer
such as LabSmith Tools for Science (CA, USA), the PEPS system could
meet the microuidic automation and control requirements for its
multi-channel output (eight-channel), wide-range voltage output
(08000 V), high-stable switch (ms-level), simple operation, compatible software with peripheral equipments, small size and cheap,
etc. The major characteristics of the PEPS system were studied by a
series of experiments.
2. Experimental
2.1. Electric units for PEPS
The main electric units for constructing PEPS system were
one single chip microprocessor (SCM) (AT89S52, Atmel Company,
USA), one communication serial port (RS-485, Taiwan Linghua
Company, China), two 12 bit digital-to-analog converters (D/A,
DAC7614, BB, USA), six ampliers (LM324, National Semiconductor, USA), eight DCDC high-voltage modules (DW-P802-1.0D,
Tianjin Dongwen Power Supply Factory, China), one input/output
(I/O) extension circuit (8255A, Intel, USA), two Darlington drivers
(ULN2803, Sprague, USA), four photocouplers (TLP521-4, Toshiba,
Japan), eight ground switch high-voltage relays (K81C235, Kilovac,

Cll Technologies, USA), eight oating switch high-voltage relays


(LRL-102-100PCV, Okita, Japan), four sampling circuits (CD4052,
TI, USA), four 12 bit analog-to-digital converters (A/D, ADS7824,
BB, USA), one over current protection relay (943H-1C-3DS, Shanghai Eighth Radio Factory, China), and one watchdog monitor circuit
(X5045, Intersil, USA). The other units such as the resistances and
capacitances were purchased from local electronic shop. Three
sizes of PEPS, the 15 cm 30 cm 26 cm, 15 cm 28 cm 24 cm
and 12 cm 22 cm 18 cm for 8000, 5000 and 2000 V voltage output, respectively, were constructed and mainly evaluated on 8000 V
voltage output in this work.
2.2. Performance evaluation of PEPS
The voltage output load of eight-electrode were experimental
performed by using the 100  standard electric resistance (SER,
Shanghai First Radio Factory, China) was connected in series with
8 M/10 kV, and 25 M/10 kV standard electric resistance (Shanghai First Radio Factory, China), respectively. As shown in Table 1,
each voltage output mode for each electrode was set as six different time intervals and repeated-running 10 times. The run time
precision, the switching time among different voltage mode and the
anti-interference ability were detected by TDS2004 oscillograph
(Tektronix, USA) connected in parallel with 100  SER. The stability
and display precision of voltage output was detected by 149-10A
digital high-voltage voltmeter (HIOKI, Japan) connected in parallel with output load. The display precision of current output was
detected by eight 44C2-A direct current micro-ammeter (Shanghai Pujiang Electrometer Factory, China) connected in series with
output load.
2.3. Microuidic chip and reagents
As shown in Fig. 1, Fig. 1a was a schematic diagram of channel design of the microuidic chip and Fig. 1b was the chip-based
CE experimental set-up used in this research. In Fig. 1a of the
glass-based microuidic chip (Dalian Institute of Chemical Physics,
Chinese Academy of Sciences, China), the injection channel, SSW,
and the separation channel, BBW, formed a cross, and the distance was 35 mm between the detection point and the cross, C. The
angle between the sample channel, SSW, and the two offset focus
channels, i.e. F1 C and F2 C, was 45 . The cross node of the two offset channels with separation channel, BBW, was in the distance of
100 m from position C. Channel cross-section was close to a rectangle structure (70 m width 25 m depth). The six reservoirs
were the same column-structure with 3 mm diameter and 1.5 mm
depth. In Fig. 1b, PEPS and chip-based CE laser induced uorescence
detector (LIFD) were homely made. Optics collection system was
confocal optics mode structure [21]. The semiconductor doublepumped solid state laser was used as the exciting light source

Table 1
Voltage output mode and time interval.
Time interval

Run time/s

Voltage output/V
V01

V02

V03

V04

V05

V06

V07

V08

1st
2nd
3rd
4th

2.5
5.0
10
20

1
300
1000
3000

1
300
1000
3000

10
500
1500
3500

10
500
1500
3500

50
700
2000
4000

50
700
2000
4000

100
900
2500
4500

100
900
2500
4500

5th

40
60

5000
Off

5000
Off

5500
Off

5500
Off

6000
Off

6000
Off

7000
Off

7000
Off

6tha

60
60

8000
G

8000
G

8000
G

8000
G

8000
G

8000
G

8000
G

8000
G

Voltage output mode is switch-off (off) switch-on (on) grounded (G).

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267

Microuids manipulations were carried out according to the


following steps. The microchannels of the chip were rstly rinsed
with 1 M NaOH for 15 min, de-ionized water for 15 min, and BBS
for 10 min; Then, 10 L BBS were lled into the reservoirs of B, F1 ,
F2 , BW and SW, respectively, and 10 L sample solution was lled
into the sample reservoir, S; and then placed the chip on the LIFD
worktable, regulated the three dimensions manipulator, observed
by CCD to focus the laser bean at 35 mm position in the separation channel near the injection cross, C. Six Pt-electrodes randomly
chosen from PEPS were inserted into the corresponding reservoirs
according to Table 2 (refer Fig. 1b). The sample injection (the sample
uid entered from reservoir, S, to the sample waste reservoir, SW),
separation (the sample uid direction was changed to ow into the
separation channel, CBW, by switching the PEPS) and sample volume were manipulated by regulating the voltage output of the PEPS
for each reservoir. The optimal voltage output applied in each reservoir in the FITC sample injection and separation was obtained by
the CCD observation of sample ow shape and direction at position,
C, and the LIFD detection sample volume of focusing voltage. While
the PEPS was operated to control the sampling and the CT-22 data
acquisition card trigger of the LIFD was operated at the same time.
Finally, the continuously four times repeated sample injection and
separation of FITC (1.0 105 M) by using PEPS and LIFD, according to the voltage program in Table 2, were carried out to evaluate
the performance of PEPS for continuously delivery and controlling
microuids in the microchannel network.
3. Results and discussion
3.1. PEPS design and construction
Fig. 1. Schematic diagrams of (a) channels design of the microuidic chip and (b) the
chip-based CE experimental set-up. S, sample reservoir; SW, sample waste reservoir;
B, buffer reservoir; BW, buffer waste reservoir; F1 and F2 , focus reservoir.

(SDPSS, MBL-20, ex = 473 nm, Changchun Xinchanye Guangdianjishu Co. Ltd., China); emission light lter was narrowband lter
with the wavelength in the range of 525 10 nm (Omega Optical,
Brattleboro, VT, USA); photomultiplier (PMT, R928, Hamamatsu,
Japan) was used as the uorescence detector; refrigeration digital
camera CCD (DS-5Mc-U2, Nikon, Japan) was used for the imaging
observation. The sampling frequency of CT-22 data acquisition card
(Shanghai Qianpu Shuju Co. Ltd., China) was 20 Hz. The connecting interface between the PEPS and the chip was also shown in
Fig. 1b, by dipping the 6-Pt-electrode (0.1 mm) of the PEPS into the
six-reservoir of the microuidic chip, respectively.
A stock solution of uorescein isothiocyanate (FITC, Sigma
Aldrich) was rst prepared in borax buffer solution (BBS,
20 103 M, pH 9.2) as 1.0 103 M, and then diluted to be
1.0 105 M sample solution by BBS. The BBS (20 103 M, pH 9.2)
was used as the running buffer solutions. All chemicals were of analytical reagent grade and doubly distilled water (Milli-Q, Millipore)
was used throughout. All the solutions were prepared by ltrating through membranes with 0.22 m sieve. The experiments were
performed at room temperature.

3.1.1. Overall design


The overview design of PEPS system was shown in Fig. 2, which
consisted of an AT89S52 SCM as a central control unit [22] and
a personal computer (PC) as an upper computer connected with
other periphery integrants. The two-directional data communication was realized through RS-485 serial port interfaces between
SCM and PC and the communication baud rate was 19,200 bit/s.
Independently eight-path-electrode voltage output was realized by
connected SCM in sequential with D/A, amplier, DCDC module
and oating/grounded relay switch touch. The switch of different
voltage output mode for each electrode was realized through initialize voltage parameter by extension circuit, Darlington driver and
photocoupler to control the switch of high-voltage relays between
grounded and oating. The display circuit was realized by connected in sequential the electrode voltage output, current (DCDC
module offer itself) with amplier, sampling circuit, A/D circuit and
SCM. Over current protection circuit was constructed by linking
the SCM with Darlington driver, over current protection relay and
DCDC power supply. When the current output monitoring value
for one or several electrodes was over the preset overload current
value, the DCDC voltage supply circuit was quickly cut off to
avoid the damage to apparatus from the overload current under the
control of SCM circuit in less than 15 ms. Finally, the working condition of SCM circuit was monitored and protected by watch-dog
circuit.

Table 2
Typical voltage output program for sample injection and separation.
Time section

1st (injection)
2nd (separation)
a

Switch-off.

Run time/s

20
70

Voltage applied/V
V01 (S)

V02 (SW)

V03 (B)

V04 (BW)

V07 (F1 )

V08 (F2 )

400
1400

0
1400

250
2000

380
0

240
Offa

240
Offa

268

Q. Li et al. / Sensors and Actuators B 136 (2009) 265274

Fig. 2. Overview of PEPS system.

The reported design took full usage of software for controlling the PEPS system, the process and the display functions of PC.
All the operations and displays would be completed within the
PC display windows, and only an on/off and a reset button were
remained in the exterior of power supply. The reported design
improved the instrumental stability, reduced the usage of hardware
and decreased the power supply and instrumental size.
3.1.2. Electrode voltage generation
The generating of electrode voltage was schematically shown
in Fig. 3. The preset voltage parameters were input through the
keyboard of the PC, and the input value was memorized into the
random access memory (RAM). The input value was transformed
to be binary numeral (for example, 110101100110) according to the
preset voltage range of PEPS program software, and the number was
transferred to SCM by serial port. Then, the numeral memorized in
the RAM was transferred to the D/A. Based on the reference voltage
value, D/A transformed the number to be relevant standard 05VDC
simulate signal at s level. The signal could control the DCDC module to output the corresponding voltage (Vo ) at ms level after it
was adjusted using amplier (equal to the voltage input value from
keyboard). These DCDC converters modules used an input voltage range between 0 and +5 V and voltage output between 0 and
+8000 V and a maximum current output of 1000 A. These specications are able to meet the power requirements at the microchip
scale.

3.1.3. Voltage output mode


As shown in Fig. 4, Fig. 4a was schematic diagram of the switch of
voltage output mode for each electrode, and Fig. 4b was schematic
diagram of the time sequence for SCM controlling the voltage output.
In Fig. 4a, J and K were grounded and oating high-voltage
relays within the power supply, respectively. J was the continuous
opening/shutting switch relay and K was the continuous opening
switch relay. The electrode was switched onto the positive port, +,
of DCDC module and the electrode voltage output was switched
on standby, when the continuous opening switch (1, 2) of K and
the continuous shutting switch (3, 2) of J were both switched on;
The electrode was switched onto the negative port, , of DCDC
module and the electrode voltage output was 0 V (grounded), when
the continuous opening switches (1, 2) of K and (1, 2) of J were both
switched on; and electrode was on the status of switch-off/oating
when the continuous opening switch (1, 2) of K was switched off.
In Fig. 4b, the voltage output mode for each electrode (switchoff/oating, on and grounded) was set according to time
section and the oating/grounded relays was controlled and
switched by SCM program. Here, it is important to prevent the
interfere of the grounded/oating switching relays from highvoltage switching, and the ground switching relays switched
50 ms behind schedule preset and oating switching relays
switched 50 ms ahead of schedule. Output time section indicated the control procedures for different operations such as

Fig. 3. Schematic diagram of the electrode voltage generation.

Q. Li et al. / Sensors and Actuators B 136 (2009) 265274

269

Fig. 4. (a) Schematic diagram of voltage output mode for each electrode; (b) time sequence for SCM controlling the voltage output. Vo , voltage output; DCDC, high-voltage
module; W1 W2 , W3 W4 , control winding of grounded and oating of voltage switching relays, respectively; VIN , input voltage of DCDC; VJ , VK voltage control of W1 W2 ,
W3 W4 , respectively.

sample injection, separation and detection during microuidic


electric-driven analysis. The automatic output sequence for each
electrode was rst time section (switch-off/run time on/run
time grounded/run time) second time section (switch-off/run
time on/run time grounded/run time), and so on. When a
voltage mode need not be output, the parameters of corresponding
mode were not set or its run time was set as 0 and then the next
mode was automatically output jumping over the set voltage mode.

3.1.4. Display output of voltage and current


The display output of voltage and current was schematically
shown in Fig. 5. The real-time voltage output monitoring value
was implemented by transforming the DCDC module from the
higher voltage to the corresponding proportional lower voltage signal using a resistance R1 (set inside of the DCDC module), and
the signal was transferred to A/D for transition by amplier 1 and
sampling circuit. The real-time current output monitoring value

Fig. 5. Schematic diagram of display of voltage output and current output.

270

Q. Li et al. / Sensors and Actuators B 136 (2009) 265274

Table 3
The experimental results for PEPS output.
Electrode
output

Voltage
range (V)

Current
range (A)

Voltage
stability (%)

Run time
precision (s)

V01
V02
V03
V04
V05
V06
V07
V08

08002
08003
08008
08007
08006
08006
08005
08003

0999
0999
0999
0999
0999
0999
0999
0999

0.025
0.038
0.1
0.088
0.075
0.075
0.063
0.038

0.097
0.096
0.099
0.098
0.097
0.093
0.096
0.091

Voltage mode switch time (ms)a


Off

On

13
12
12
11
13
14
14
13

47
46
48
44
47
48
49
43

9.7
9.2
10
9.1
10
10
9.8
10

Voltage display
error (%)
0.13
0.14
0.16
0.18
0.19
0.17
0.15
0.17

Current display
error (%)
0.53
0.55
0.57
0.58
0.59
0.56
0.54
0.59

Off, switch-off; On, switch-on; G, grounded.

was implemented by transforming the current passing through the


sampling resistance R2 (set inside of the DCDC module) in the
output circuit to the corresponding voltage signal, and the signal
was transferred to A/D for transition by amplier 2 and sampling
circuit. The two rheostats of W1 and W2 were used to inching regulate the two ampliers, amplier 1 and amplier 2 (to make the
output value equal to the display value). After being memorized by
the RAM of SCM, the value completed from the A/D module was
transmitted to CPU for logic process and real-time displayed on PC.
3.1.5. Program software
The program software for PEPS system consisted of a SCM control program and a PC application program, which were compiled
by assembly language and C++ [23], respectively. The SCM control program could realize the function of date processing, over
current protection, time sequence control between the circuits,
series/parallel drive, 12 bit D/A, 12 bit A/D and I/O bus-mastering,
etc. The PC application program included multiple operation interfaces such as serial port initializing, new experiment setting up,
voltage parameter setting (adjustable voltage ranges of 2000, 5000
and 8000 V for each electrode, and 8000 V was chosen in this work),
voltage parameter presetting, parameter application and running,
real-time data/curve displaying, etc. The PC application program
could be operated under Windows98/2000/XP with friendly windows. When the experiment was ended, the program software
would save automatically all the parameters set in the full running
process in CE operation including electrode number, electrode voltage output mode, running time, over current protection value and
repeated running times and so on, and these parameters would not
be set again under the same experimental conditions. This software
could be compatible with multiple application programs under
Windows system and all the chip-based electrophoresis detectors
operated under Windows system could be coupled with the PEPS
system for a certain usage.
3.1.6. Anti-interference strategy
In multi-channel voltage sequencer, it is easy to give rise to
interference due to the increased numbers of the voltage output
channels. The possible interference of the voltage output exists in
both the control system and channel-to-channel of the voltage outputs. The main strategies of PEPS to solve the interference problem
and to make sure a high-stable switch were listed as follows:
(1) use the photoelectron coupling technique to separate the SCM
and high-voltage relays to avoid interferences between the
modules;
(2) optimize the grounded wire distribution reasonably and separate the digital grounded and the analog grounded to avoid the
interference of the digital signals on the analog signals;

(3) add de-dipole capacitance at the input power supply of the PEPS
system to decrease high-frequency interferences;
(4) avoid the grounded/oating switching relays from highvoltage switching (Fig. 4b) and cancel the onoff switching
interferences;
(5) use a watch-dog circuit to avoid the SCM from status of a dead
running cycle and/or a departure and to prevent SCM from
out of control status that possibly lead illegal operation of the
controlled bit;
(6) moreover, optimize the distribution of the electric board, for
example, to separate the SCM and relay as far as possible that
decrease the conduct interference.
3.2. Evaluation of PEPS
The experimental results for PEPS system were shown in Table 3.
The results indicated that the experiment parameters such as
electrode path (18 path), voltage output mode for each electrode (switch-off/oating, on and grounded), time section
(16 section), run time (09999 s) and repeated running time
(09999 times) could be randomly preset as desired. The voltage
output of the eight electrodes could be operated synchronically
according to time section preset and the voltage mode for each electrode could be automatically controlled and quickly transformed
without any interference. For each electrode, the parameters such
as the status of voltage mode, the voltage output, curves of current output vs time were all real-time displayed at the same time.
When the output load of resistance changed with 200%, the voltage output and current output were in the range of 0 to +8000 V
and 0 to +999 A, respectively. The precision of voltage output and
the run time for PEPS were less than 0.1% and 0.1 s, respectively,
the display precision of voltage output and current output were
less than 0.2% and 0.6%, respectively, and the switching time of
switch-off/oating, on and grounded output mode was less
than 15 ms, 50 ms and 10 ms, respectively.
The characteristics such as eight electrodes (operated independently for single electrode or output synchronously for multiple
electrodes combination), programmable control the voltage output
and running time, and output stable, indicated that the PEPS system
could conveniently and automatically control the ow rate and the
direction of uids in the microchannel network, which could be
easily realized by randomly regulating the electric eld strength
and direction applied in the channel network.
3.3. PEPS applied for chip CE
3.3.1. Choice of electrode voltage
The optimal voltage output of the electrode applied in the reservoirs could control the process of chip CE injection and separation
with high efciency [24]. The pinched sample injection, electroki-

Q. Li et al. / Sensors and Actuators B 136 (2009) 265274

271

According to Eqs. (1) and (2), the voltage on cross node C, VC , is


VC =

VB /RB + VF1 /RF1 + VS /RS + VF2 /RF2 + VBW /RBW + VSW /RSW
1/RB + 1/RF1 + 1/RS + 1/RF2 + 1/RBW + 1/RSW
(3)

and assumed that the cross-sectional area and the electrolyte in the
channel are the same, and Eq. (3) is
VC =

VB /LB + VF1 /LF1 + VS /LS + VF2 /LF2 + VBW /LBW + VSW /LSW
1/LB + 1/LF1 + 1/LS + 1/LF2 + 1/LBW + 1/LSW
(4)

The electric eld strength, Ei , in each channel is


Ei =

Vi VC
Li

(5)

The electroosmotic velocity, vieo , electrophoresis velocity, viep , and


apparent electromigration velocity, vi , of the sample in each channel
can be expressed as [27]

vieo = eo Ei =

eo (Vi VC )
Li

(6)

viep = ep Ei =

ep (Vi VC )
Li

(7)

vi = vieo + viep = (eo + ep )Ei =

Fig. 6. (a) Schematic diagram of voltages applied to the six reservoirs of the glassbased chip at injection step and the movement of EOF. (b) EER of microchannel
network. Thin lines, the external electrical connections and applied voltages; S,
sample reservoir; SW, sample waste reservoir; B, buffer reservoir; BW, buffer waste
reservoir; F1 , F2 , focus reservoir.

netic focusing injection method was used in this experiment [25]


to demonstrate the applicable of the PEPS system. In order to obtain
the ideal results in injection and separation steps, the circuit analysis and the CCD observation had been used for evaluating the uid
shape and the uid direction in the cross, C. When the voltages
applied to the six reservoirs and the equivalent electric resistance
(EER) of microchannel network were shown in Fig. 6.
Based on the Ohm Law, the current, Ii , of each channel in Fig. 6b
is
Ii =

Vi VC
Ri

(1)

where Vi is the applied voltage in each reservoir, VC is the voltage


on cross node C, Ri (Ri = Li /kAi ) is the EER of the electrolyte solution
between each reservoir and C, of each channel [26]. In Ri = Li /kAi ,
Li is the length between each reservoir and C of each channel, k is
the conductance of the electrolyte solution in the channel, Ai is the
cross-sectional area of each channel, and i is B, F1 , S, F2 , BW and
SW, respectively. Based on the Kirchhoff Current Law, the current
on cross node C, IC , is
IC = IB + IF1 + IF2 + IS + IBW + ISW = 0

(2)

(eo + ep )(Vi VC )


Li

(8)

where eo is electroosmosis mobility of electrolyte solution


(eo = /, where is dielectric permittivity,  is zeta potential in
the electrical double layer (EDL) of liquidsolid interface,  is viscidity); and ep is electrophoresis mobility of sample (ep =  e /,
where  e is the zeta potential of the sample). The magnitude of
the eo is normally much larger than the ep of charged species
(however, there are some exceptions to this rule), and eo is always
larger than ep in glass-based materials at pH >6.0 [6]. In this work,
the experimental conditions, including the capillary zone electrophoresis (CZE), glass-based chip and pH 9.2 (BBS), the sample
molecules of FITC could be assumed as spherical particles with partial negative charges, and the ep /eo was estimated about 3.89%.
The EOF was dominant in this experiment. So, the ow velocity of
sample is similar to EOF velocity with minor difference [28].
In the sample injection step, as shown in Fig. 7a, the ideal
pinched injection could be obtained when set the voltage of
sample reservoir, S, as 400 V, sample waste reservoir, SW, as
grounded, buffer reservoir, B, as 250 V, buffer waste reservoir, BW,
as 360400 V, the focus voltage of F1 and F2 as both 240 V, and
voltage on the cross, C, as about 230 V (according to Eq. (4)). In the
sample separation step, as shown in Fig. 7b, the satised separation was obtained when set the voltage of S, B, SW and BW as 1600,
2000, 1600 and 0 V, respectively, and kept F1 and F2 as oating.
However, when injection was repeatedly only one time the sample solution would leak out. When the applied voltage of S and SW
was decreased to 1400 V, the above problem could be solved. The
series manipulation of the uids sample volume formation, sample zone stabilization and sample un-leakage can be realized with
easy by six channels of the PEPS system. The apparent electromigration velocity (separation channel, from cross to detection point) is
estimated about 0.080 cm/s by measuring the migration distance
of uorescent band in a given time period using the sequential
images. This is further proved by the chip-based electrophoresis
experiments in Fig. 9.
3.3.2. Control of sampling volume
In the sample injection step (Figs. 1a and 7), the sampling
volume was controlled by regulating the focusing voltage value
applied on the reservoirs, F1 and F2 , on two offset microchannels.

272

Q. Li et al. / Sensors and Actuators B 136 (2009) 265274

Fig. 7. The sample streamline in cross, C, using pinched injection method. (a) Injection step; (b) separation step.

The preset parameters of PEPS system for controlling FITC chipbased CE operations such as injection, separation were
(1) injection step, the voltages value applied on reservoirs of S, SW,
B and BW (V01 , V02 , V03 , V04 ) was kept invariable according to
the program listed in Table 2, and the focusing voltage value of
F1 and F2 (V07 , V08 , V07 = V08 ) was set as 240, 260, 280 and 300 V,
respectively; In this work, the volume of cross C was estimated
as 140.6 pL, as shown in Fig. 7a, the sampled volume was one
third of the above volume, i.e. 46.9 pL;
(2) separation step, the voltage value applied on reservoirs of S, SW,
B, BW, F1 and F2 was kept invariable according to the program
listed in Table 2.
The results detected by LIFD were shown in Fig. 8. From Fig. 8a
and b, the voltage change applied on the reservoirs of two offset
channel could affect the sample volume and nal sample peak (peak
height and peak area). The reason of the focus voltage affect the
sample volume was that the focused voltage change would change
the ow rate and thereby change the volume ratio as well the ow
shape of the two phases (the running buffer and the sample solution) [29], and further change the stored sample volume in the
cross, C, and nally the width and location of sample in the sampling
channel could be controlled indirectly. The higher of the voltage
applied and the narrower sample ow. Under these conditions, the
sampling volume was the function of focused voltage, sampling
time, sampling volume (in the cross, C) and running buffer ow
rate. Hence, combining the optimal design of the microuidic structure with the precise control and steady output and run time could
adjust the sample volume. The voltage output program of the PEPS

system in Table 2 was used for controlling the operation of the FITC
CE sample injection and separation based on above observation and
the detection of LIFD.
3.3.3. Continuous control injection/separation operation
According to the voltage program listed in Table 2, PEPS system was used for continuously controlled FITC injection/separation
operation four times and detected by LIFD, the results were shown
in Fig. 9. We could conclude that the voltage with synchronous
switch and steady output of PEPS system in each reservoir (S, SW,
B, BW, F1 and F2 ) can automatically realize the continuous injection and separation of chip-based CE operation, and also ensure
the relative standard deviation (R.S.D., n = 4, S/N = 10) of the analysis results with 0.90% for the reservation time (tR ) and 2.3% for the
peak height, respectively.
The main reason for this was, in the sample injection step,
the voltage in the reservoirs (S, B, BW, F1 and F2 ) was all
higher than that at the cross node, C (about 230 V according
to Eq. (4)) and the voltage in the sample waste reservoir, SW,
was the lowest (V02 = 0 V, grounded). The distribution of electric eld strength between the six reservoirs and cross, C, was:
ECSW > ESC > EBC /EBWC > EF1C /EF2C . Hence, the conuent ows
of the major ow from the sample reservoir, S, and minor ows
running buffer reservoirs, F1 and F2 , forming three-layer parallel
laminar ow, and then the uid owed into SW after passing the
cross; the buffer from B and BW compressed the sample ow at
the cross, C, which reduced the pervasion accumulation when the
sample was injected into the separation channel (Fig. 7a). It should
be pointed out that the liquid at lower ow velocity in microchannel formed a laminar ow, and the advection effect at the interface

Q. Li et al. / Sensors and Actuators B 136 (2009) 265274

273

voltage in the reservoirs of sample, S, and sample waste, SW, was


lower than that in the cross node, C, and the voltage in the reservoir
of the buffer waste, BW, was the lowest (V04 = 0 V, grounded) during
which the distribution of electric eld strength at each reservoir and
cross node, C, was EBC > ECBW > ECS /ECSW . The buffer ow from
reservoir, B, pushed the sample at the cross, C, to the separation
channel (CBW), and formed sample section (Fig. 7b); at the same
time a small amount of running buffer ow at low ow rate from
the analysis channel, BC, entered the sample injecting channel
border upon the cross node so that the sample in the sample channel owed backwards (Fig. 7b) to decrease or avoid the interference
on the electrophoresis analysis effect when the sample leaks entering the separation channel. As a result, the detection background
signal decreased and baseline became steady (Fig. 9). It is obvious
that the other sample injection modes of the chip-based CE will
be easy obtained by using PEPS system and setting the voltages
distribution on the reservoirs, which have been demonstrated by
CE analysis using PEPS for different chip-based network as well as
different-type detections [28,3133].
4. Conclusion

Fig. 8. (a) Inuence of focusing voltage on sample volume and corresponding sample peak; (b) variations of sample peak height and sample peak area vs focusing
voltage.

between the sample and running buffer ow can be ignored at a


short-time period [30]. In the separation step, i.e. the step of changing the direction of the sample, the voltage in the reservoir, B, of
running buffer was higher than that at the cross node, C, while the

In this paper, a versatile programmable PEPS system was


constructed and evaluated. The performances such as capability, accuracy and programmability of the developed PEPS system
were investigated and evaluated in detail through the injection
and separation microuidic experiments, including voltage output generation, and voltage output mode (oating/switch-off, on
and grounded), running time. The presented system could control the connection among the six-reservoir and manipulate the
microuids in any direction and the ow rate as desired using
the system program software. PEPS system gave good performance
and met the requirements sufciently for microuidic applications
especially multichannel chip network (for example, ve to eight
reservoirs). The convenience, stable, and simple of the PEPS system
will nd its potential applications in controlling and manipulating
the microuids in the future.
Acknowledgements
This work was supported by Natural Science Foundation of China
(Nos. 20475034 and 20875058), National Basic Research Program
of China (973 Program, 2007CB936000), Natural Science Funds
for Distinguished Young Scholar of China (No. 20725518), Major
Program of Natural Science Foundation of China (No. 90713019),
Natural Science Foundation of Shandong Province in China (No.
Y2008B15).
References

Fig. 9. Electropherogram for continuously injection-separation operation. Chip,


glass-based microuidic chip; sample concentration, 1.0 105 M FITC; running
buffer concentration: 20 105 M BBS (pH 9.2); detector, LIFD at ex = 473 nm,
em = 525 nm.

[1] G.M. Whitesides, The Origins and the future of microuidics, Nature 442 (2006)
368373.
[2] P.S. Dittrich, K. Tachikawa, A. Manz, Micro total analysis systems. Latest
advancements and trends, Analytical Chemistry 78 (2006) 38874908.
[3] A. Wolff, I.R. Perch-Nielsen, U.D. Larsen, P. Friis, G. Goranovic, C.R. Poulsen,
J.P. Kutter, P. Telleman, Integrating advanced functionality in a microfabricated
high-throughput uorescent-activated cell sorter, Lab on a Chip 3 (2003) 2227.
[4] D.J. Laser, J.G. Santiago, A review of micropumps, Journal of Micromechanics
and Microengineering 14 (2004) R35R64.
[5] L.X. Chen, J. Choo, B. Yan, Microfabricated electrokinetic ow pump and its
related techniques: a potential promising drug delivery technique, Expert Opinion on Drug Delivery 4 (2007) 119129.
[6] L.X. Chen, S. Lee, J. Choo, E.K. Lee, Continuous dynamic ow micropumps for
microuid manipulation, Journal of Micromechanics and Microengineering 18
(2008) 013001.
[7] J. Jung, L.X. Chen, S. Lee, S. Kim, G.H. Seong, J. Choo, E.K. Lee, C.H. Oh, S. Lee,
Fast and sensitive DNA analysis using changes in the FRET signals of molecular
beacons in a PDMS microuidic channel, Analytical and Bioanalytical Chemistry
387 (2007) 26092615.

274

Q. Li et al. / Sensors and Actuators B 136 (2009) 265274

[8] M.A. Unger, H.P. Chou, T. Thorsen, A. Scherer, S.R. Quake, Monolithic microfabricated valves and pumps by multilayer soft lithography, Science 288 (2000)
113116.
[9] J.W. Munyan, H.V. Fuentes, M. Draper, R.T. Kelly, A.T. Woolley, Electrically actuated, pressure-driven microuidic pumps, Lab on a Chip 3 (2003)
217220.
[10] S. Kar, S. McWhorter, S.M. Ford, S.A. Soper, Piezoelectric mechanical pump with
nanoliter per minute pulse-free ow delivery for pressure pumping in microchannels, Analyst 123 (1998) 14351441.
[11] J. Atencia, D.J. Beebe, Magnetically-driven biomimetic micro pumping using,
Lab on a Chip 4 (2004) 598602.
[12] L.X. Chen, J.P. Ma, F. Tan, Y.F. Guan, Generating high-pressure sub-microliter ow
rate in packed microchannel by electroosmotic force: potential application in
microuidic systems, Sensors and Actuators B 88 (2003) 260265.
[13] L.X. Chen, H.L. Wang, J.P. Ma, C.X. Wang, Y.F. Guan, Fabrication and characterization of a multi-stage electroosmotic pump for liquid delivery, Sensors and
Actuators B 104 (2005) 117123.
[14] B. Anders, S. Detlef, P.K. Jrg, B. Henrik, AC electroosmotic pump with bubblefree palladium electrodes and rectifying polymer membrane valves, Lab on a
Chip 6 (2006) 280288.
[15] P. Woias, Micropumpspast, progress and future prospects, Sensors and Actuators B 105 (2005) 2838.
[16] E.S. Roddy, H.W. Xu, A.G. Ewing, Sample introduction techniques for microfabricated separation devices, Electrophoresis 25 (2004) 229242.
[17] C.D. Garca, Y. Liu, A. Paul, C.S. Henry, Versatile 3-channel high-voltage
power supply for microchip capillary electrophoresis, Lab on a Chip 3 (2003)
324328.
[18] P.D.I. Fletcher, S.J. Haswell, X.L. Zhang, Electrokinetic control of a chemical
reaction in a lab-on-a-chip micro-reactor: measurement and quantitative modeling, Lab on a Chip 2 (2002) 102112.
[19] Z.Y. Wu, N. Xanthopoulos, F. Reymond, J.S. Rossier, H.H. Girault, Polymer microchips bonded by O2 -plasma activation, Electrophoresis 23 (2002)
782790.
[20] Q.L. Li, B. Tang, H.X. Tian, Chinese Patent 200510104343.3, 2005.
[21] X.M. Zhou, D.Y. Liu, Y.T. Zhong, Z.P. Dai1, D.P. Wu, H. Wang, Y.G. Du, Z.A. Xia, L.P.
Zhang, X.D. Mei, B.C. Lin, Determination of SARS-coronavirus by microuidic
chip system, Electrophoresis 25 (2004) 30323039.
[22] Y.C. Sun, Application of AT89 Catena Single Chip Microcomputer, Tsinghua University Press, Beijing, 2005.
[23] D.S. Malik, Data Structures Using C++, Tsinghua University Press, Beijing, 2003.
[24] L.M. Fu, C.H. Lin, Numerical analysis and experimental estimation of a lowleakage injection technique for capillary electrophoresis, Analytical Chemistry
75 (2003) 57905796.
[25] L.M. Fu, R.J. Yang, G.B. Lee, Electrokinetic focusing injection methods on
microuidic devices, Analytical Chemistry 75 (2003) 19051910.
[26] P.D.I. Fletcher, S.J. Haswell, X. Zhang, Electrical currents and liquid ow rates in
micro-reactors, Lab on a Chip 1 (2001) 115121.
[27] M.B. Christopher, H.D. Robert, Electroosmotic ow in channels with step
changes in zeta potential and cross section, Journal of colloid and Interface
Science 270 (2004) 242246.
[28] L.X. Chen, G.A. Luo, K.H. Liu, J.P. Ma, B. Yao, Y.C. Yan, Y.M. Wang, Bonding of glassbased microuidic chips at low- or room-temperature in routine laboratory,
Sensors and Actuators B 119 (2006) 335344.

[29] P.J.A. Kenis, R.F. Ismagilov, G.M. Whitesides, Microfabrication inside capillaries
using multiphase laminar ow patterning, Science 285 (1999) 8385.
[30] B.H. Weigl, P. Yager, Microuidicsmicrouidic diffusion-based separation and
detection, Science 283 (1999) 346347.
[31] B. Ma, X.M. Zhou, G. Wang, H.Q. Huang, Z.P. Dai, J.H. Qin, B.C. Lin, Integrated
isotachophoretic preconcentration with zone electrophoresis separation on
a quartz microchip for UV detection of avonoids, Electrophoresis 27 (2006)
49044909.
[32] W. Wang, X.Q. Xu, Q. Bin, J.M. Ling, G.N. Chen, A new method for fabrication of
an integrated indium tin oxide electrode on electrophoresis microchips with
amperometric detection and its application for determination of synephrine
and hesperidin in pericarpium citri reticulatae, Electrophoresis 27 (2006)
41744181.
[33] B. Ma, X.M. Zhou, G. Wang, Z.P. Dai, J.H. Qin, B.C. Lin, A hybrid microdevice with
a thin PDMS membrane on the detection window for UV absorbance detection,
Electrophoresis 28 (2007) 24742477.

Biographies
Qingling Li received his B.Sc. in electrical engineering from Shandong Industrial
University in 1984. From 1984 to 2005, he was an engineer and senior engineer in
Shandong Chemical Engineering Institute. Since 2005, he has been a senior engineer in College of Chemistry at Shandong Normal University. His research interests
include capillary electrophoresis, microuids delivery & control and microuidic
detection.
Hui Zhang received her B.Sc. in chemistry education from Shandong Normal University in 2001, and a M.Sc. in analytical chemistry from Shandong Normal University
in 2004. Now, she is a Ph.D. candidate in Shandong Chemical Engineering Institute.
Her research interest is focused on ow injection analysis.
Yan Wang received her B.Sc. and M.Sc. in optical analytical chemistry from Shandong
University in 1990 and in 1993, respectively. Since 1993, she is an associate professor
in College of Chemistry at Shandong Normal University. Her research interest is
focused on capillary electrophoresis.
Bo Tang received his B.Sc. degree in Applied Chemistry from China University of
Geosciences in 1988, and a M.Sc. and a Ph.D. in optical analytical chemistry from
Nankai University in 1991 and in 1994, respectively. Since 1994, he is an associate
professor and professor in College of Chemistry at Shandong Normal University. His
research interests include optical analysis, uorescence nanoprobe and molecular
identication.
Xin Liu received her B.Sc. in Environment Engineering from North University of
China in 2006. Since 2006, she is a Ph.D. candidate in College of Chemistry at Shandong Normal University. Her research interests include microchip electrophoresis
and microuidic detection.
Xiaocong Gong received her B.Sc. in chemistry education from Linyi Normal University in 2006. Since 2006, she is a graduate student in College of Chemistry at
Shandong Normal University. Her research interest is focused on micrototal analysis.

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