Download LONG-TERM CELL IMAGING IN A MICROCHAMBER WITH

LONG-TERM CELL IMAGING IN A MICROCHAMBER WITH VARIABLLY
CONTROLLED CONCENTRATION GRADIENT GENERATION
Ji-Yen Cheng1,*, Meng-Hua Yen1,2, Ching-Te Kuo1, Tai-Horng Young2
Research Center for Applied Sciences, Academia Sinica Taiwan
2
Institute of Biomedical Engineering, National Taiwan University, Taiwan
E-mail: [email protected]
KEY WORDS: long-term cell imaging, concentration gradient, microfluidic chamber,
chemotaxis, electrotaxis
1
1. INTRODUCTION
We report a micro device suitable cell culturing on a microscope so that long-term
observation on cell behavior, e.g. chemotaxis, can be performed outside of an incubator.
The device consists of an integrated transparent heater and a variably controlled
concentration gradient generator. Successful culturing was performed inside the device.
Continuous long-term (> 10 days) observation on cell growth was achieved. In this work,
the flow field, medium replacement, and chemical gradient build-up in the microchamber
and its performance as an independent cell culture device is elaborated.
2. EXPERIMENT AND RESULT
Microfluidic chamber and transparent ITO heater were fabricated by our laser ablation
technologies(1,2). The microchamber consists of three connectors (figure 1(a)). Fresh
medium was introduced via left connector and waste out via the right connector. The
center connector is for introducing chemical attractant (or repellant) of cell so that
concentration gradient was generated inside the cell culture chamber (figure 1(b). By
controlling the relative pumping ration between the medium flow and center chemical
flow the concentration gradient can be changed (figure 19(c)). Such device is very useful
for long-term cell behavior studies, e.g. chemotaxis and electrotaxis. A microscopic
image of cells cultured for 122 hours in this device is shown in figure 1(d).
(a)
(b)
(c)
(d)
Figure 1. (a) Photo picture of the cell culturing device attached on a microscope. (b)
Concentration gradient generated by 10 μL/h lateral medium flow and 10 μL/h center chemical
flow. (c) Concentration gradient generated by 50 μL/h medium flow and 10 μL/h center flow. (d)
Microscopic picture of living CL1-5 lung cancer cell cultured after 122 hours in the device.
References:
1. Cheng, J.-Y., Wei, C.-W., Hsu, K.-H. and Young, T.-H. (2004) Direct-write Laser Micromachining and
Universal Surface Modification of PMMA for Device Development. Sensors and Actuators B:Chemistry,
99, 186-196.
2. Cheng, J.-Y., Yen, M.-H., Hsu, W.-C., Jhang, J.-H. and Young, T.-H. (2007) ITO patterning by a low
power Q-switched green laser and its use in the fabrication of a transparent flow meter. J. Micromech.
Microeng., 17, 2316-2323.