A microfluidic platform for sequential ligand labeling and cell binding analysis.
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.
Sui
Guodong
G
Department of Molecular Medical Pharmacology and Crump Institute for Molecular Imaging, University of California, Los Angeles, 700 Westwood Plaza, Los Angeles, CA 90095, USA.
Lee
Cheng-Chung
CC
Kamei
Ken-Ichiro
K
Li
Hua-Jung
HJ
Wang
Jin-Yi
JY
Wang
Jun
J
Herschman
Harvey R
HR
Tseng
Hsian-Rong
HR
D000818:Animals; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods; D002462:Cell Membrane / Q000378:metabolism*; D004815:Epidermal Growth Factor / Q000378:metabolism*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D066246:ErbB Receptors / Q000378:metabolism*; D005434:Flow Cytometry / Q000295:instrumentation* / Q000379:methods; D017022:Flow Injection Analysis / Q000295:instrumentation / Q000379:methods; D008024:Ligands; D051379:Mice; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D041681:NIH 3T3 Cells; D025941:Protein Interaction Mapping / Q000379:methods*; D013194:Staining and Labeling / Q000379:methods
D008024:Ligands; D004815:Epidermal Growth Factor; D066246:ErbB Receptors