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Anal. Chem. 2004, 76, 483-488

Transport, Location, and Quantal Release
Monitoring of Single Cells on a Microfluidic Device
Wei-Hua Huang,† Wei Cheng,† Zhen Zhang,† Dai-Wen Pang,† Zong-Li Wang,† Jie-Ke Cheng,*,† and
Da-Fu Cui‡

Department of Chemistry, Wuhan University, Wuhan 430072, China, and State Key Laboratories of Transducer Technology,
The Institute of Electronics, Chinese Academy of Science, Beijing 100080, China

A novel microfluidic device has been developed for onchip transport, location, and quantal release monitoring
of single cells. The microfluidic device consists of a plate
of PDMS containing channels for introducing cells and
stimulants and a glass substrate into which a cell microchamber was etched. The two tightly reversibly sealed
plates can be separated for respective cleaning, which
significantly extends the lifetime of the microchip that is
frequently clogged in cell analysis experiments. Using
hydraulic pressure, single cells were transported and
located on the microfluidc chip. After location of a single
PC12 cell on the microfluidic chip, the cell was stimulated
by nicotine that was also introduced through the microchannels, and the quantum release of dopamine from the
cell was amperometricly detected with our designed
carbon fiber microelectrode. The results have demonstrated the convenience and efficiency of using the microfluidic chip for monitoring of quantal release from
single cells and have offered a facile method for the
analysis of single cells on microfluidic devices.
Research on biological cells has been deepening into a more
diminutive scale, from cellular clusters to single cells, even to
subcellular compartments. Due to the small size of single cells
(diam 7-200 µm, vol femtoliters to nanoliters), trace amounts of
sample (zeptomoles to femtomoles), complex components, and
millisecond scale of reaction time, single-cell analysis offers a
severe challenge for development of analytical instrumentation.
Since the first assay of a single neuron from Helix aspersa by
capillary liquid chromatography and capillary electrophoresis
(CE),1 CE combining detection techniques of electrochemistry,
laser-induced fluorescence, mass spectrometry etc.2 has been
widely employed for single-cell analysis. Recently, other advanced
analytical methods involving microfluidic chip,3 imaging analysis,4
and temporal and spatial dynamic monitoring with microeletrodes5
* Corresponding author. Phone: +86-27-87682291. Fax: +86-27-87647617.

Wuhan University.

Chinese Academy of Science.
(1) Kennedy, R. T.; Oates, M. D.; Cooper, B. R.; Nickerson, B.; Jorgenson, J.
W. Science 1989, 246, 57-63.
(2) Stuart, J. N.; Sweedler, J. V. Anal. Bioanal. Chem. 2003, 375, 28-29.
(3) Auroux, P. A.; Iossifidis, D.; Reyes, D. R.; Manz, A. Anal. Chem. 2002, 74,
2637-2652.
(4) Swedlow, T. R.; Goldberg, I.; Brauner, E.; Sorger, P. K. Science. 2003,
300, 100-102.
10.1021/ac035026y CCC: $27.50
Published on Web 12/10/2003

© 2004 American Chemical Society

have also increasingly been developed and widely applied to singlecell analysis.
Because of its unique advantages, such as rapid speed and
low cost, the microfluidic device has attracted significant attention
and also has displayed its great potential in the research of singlecell analysis. The scale of microchannels in the microfluidic chip
is just fitted to the size of most cells, and the ability to decrease
both the consumption of analytes and the duration of analyses
allows high time-resolved single-cell analysis. Previous studies
have mainly focused on cell flow cytometry,6-8 sorting,9,10 fusion,11
and on-chip cell analysis involving cell lysis and consequent
intercellular reaction, for example, protein extraction,12 PCR
amplification,13 intracellular enzyme reaction,14 and intercellular
substance measurement in a cell cultured microchip.15 So far, only
a few studies have carried out the manipulation and analysis of
living cells on various functional microfluidic devices. Li and
Harrison16 first performed the manipulation and transport of
various cells, such as red blood cells, yeast cells, and E.coli,
throughout a channel network using electroosmotic and electrophoretic pumping, and demonstrated cell lysis process by SDS at
a T-junction on a silicon-based microfluidic chip. Yang et al.17 have
constructed a dam structure in a microfluidc chip for location and
alignment of cells. It allows the fragile cells to move in the
microfluidic channels and to be immobilized in controllable
numbers in the desired locations under fluid pressure. The
calcium uptake reaction of HL-60 cells was monitored after cell
(5) Huang, W. H.; Hu, S.; Pang, D. W.; Wang, Z. L.; Cheng, J. K. Chin. Sci.
Bull. 2000, 45, 289-295.
(6) McClain, M. A.; Culbertson, C. T.; Jacobson, S. C.; Ramsey, J. M. Anal.
Chem. 2001, 73, 5334-5338.
(7) Schrum, D. P.; Culbertson, C. T.; Jacobson, S. C.; Ramsey, J. M. Anal. Chem.
1999, 71, 4173-4177.
(8) Gawad, S.; Schild, L.; Renaud, Ph. Lab Chip 2001, 1, 76-82.
(9) Fiedler, S.; Shirley, S. G.; Schnelle, T.; Fuhr, G. Anal. Chem. 1998, 70,
1909-1915.
(10) Fu, A. Y.; Spence, C.; Scherer, A.; Arnold, F. H.; Quake, S. R. Nat. Biotechnol.
1999, 17, 1109-1111.
(11) Stromberg, A.; Karlsson, A.; Ryttsen, F.; Davidson, M.; Chiu, D. T.; Orwar,
O. Anal. Chem. 2001, 73, 126-130.
(12) Schilling, E. A.; Kamholz, A. E.; Yager, P. Anal. Chem. 2002, 74, 17981804.
(13) Waters, L. C.; Jacobson, S. C.; Kroutchinina, N.; Khandurina, J.; Foote, R.
S.; Ramsey, J. M. Anal. Chem. 1998, 70, 158-162.
(14) Heo, J.; Thomas, K. J.; Seong, G. H.; Crooks, R. M. Anal. Chem. 2003, 75,
22-26.
(15) Tamaki, E.; Sato, K.; Tokeshi, M.; Sato, K.; Aihara, M.; Kitamori, T. Anal.
Chem. 2002, 74, 1560-1564.
(16) Li, P. C. H.; Harrison, D. J. Anal. Chem. 1997, 69, 1564-1568.
(17) Yang, M.; Li, C. W.; Yang, J. Anal. Chem. 2002, 74, 3991-4001.

Analytical Chemistry, Vol. 76, No. 2, January 15, 2004 483


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