A microfluidic platform for sequential ligand labeling and cell binding analysis
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Developing biochemical and cell biological assay for screening biomolecules, evaluating their characteristics in biological processes, and determining their pharmacological effects represents a key technology in biomedical research. A PDMS-based integrated microfluidic platform was fabricated and tested for facilitating the labeling of ligand on the nanogram scale and sequential cell binding analysis in a manner that saves both time and reagents. Within this microfluidic platform, ligand labeling, cell immobolization, and optical analysis are performed in a miniaturized, continuous and semi-automated manner. This microfluidic device for ligand labeling and cell analysis is composed of two functional modules: (i) a circular reaction loop for fluorophore-labeling of the ligand and (ii) four parallel-oriented incubation chambers for immobilization of cells, binding of ligand to different cell populations, and optical evaluation of interactions between the labeled ligand and its cell targets. Epidermal growth factor (EGF) as the ligand and different cell lines with various levels of EGF receptor expression have been utilized to test the feasiblity of this microfluidic platform. When compared to studies with traditional Petri dish handling of cells and tissues, or even microwell analyses, experiments with the microfluidic platform described here are much less time consuming, conserve reagents, and are programmable, which makes these platforms a very promising new tool for biological studies.
KeywordsMicrofluidics Ligand labeling PDMS EGF EGFR
This research was supported by the UCLA In Vivo Cellular and Molecular Imaging Center (P50 CA086306:06), the UCLA Prostate SPORE (P50 CA092131:04) and institutional “start up” funds to HRT. GS acknowledges the support from UCLA-SOMI (Scholars in Oncologic Molecular Imaging) program. The authors thank Prof. Stephen R. Quake from Stanford University for his kind help.
- A. Aharonov, R. Pruss, and H. Herschman, J. Cell. Biol. 75, A188 (1977).Google Scholar
- R.B. Bird, W.E. Stewart, and E.N. Lighttfoot, Transport Phenomena (John Wiley & Sons, 2002).Google Scholar
- P. Nair, Curr. Sci. 88, 890–898 (2005).Google Scholar
- T. Thorsen, S.J. Maerkl, and S.R. Quake, Science 298, 580–584 (2002).Google Scholar