Isotropically etched radial micropore for cell concentration, immobilization, and picodroplet generation

• Published in: Lab on a chip 2009-01, Vol.9 (4), p.507-515
• Main Authors: Perroud, Thomas D, Meagher, Robert J, Kanouff, Michael P, Renzi, Ronald F, Wu, Meiye, Singh, Anup K, Patel, Kamlesh D
• Format: Article
• Language: English
• Subjects: Animals; Cell Line; Cytological Techniques; Equipment Design; Escherichia coli - cytology; Macrophages - cytology; Mice; Microfluidic Analytical Techniques - instrumentation; Microfluidic Analytical Techniques - methods; Porosity; Reproducibility of Results
• ISSN: 1473-0197; 1473-0189
• DOI: 10.1039/b817285d

To enable several on-chip cell handling operations in a fused-silica substrate, small shallow micropores are radially embedded in larger deeper microchannels using an adaptation of single-level isotropic wet etching. By varying the distance between features on the photolithographic mask (mask distance), we can precisely control the overlap between two etch fronts and create a zero-thickness semi-elliptical micropore (e.g. 20 microm wide, 6 microm deep). Geometrical models derived from a hemispherical etch front show that micropore width and depth can be expressed as a function of mask distance and etch depth. These models are experimentally validated at different etch depths (25.03 and 29.78 microm) and for different configurations (point-to-point and point-to-edge). Good reproducibility confirms the validity of this approach to fabricate micropores with a desired size. To illustrate the wide range of cell handling operations enabled by micropores, we present three on-chip functionalities: continuous-flow particle concentration, immobilization of single cells, and picoliter droplet generation. (1) Using pressure differentials, particles are concentrated by removing the carrier fluid successively through a series of 44 shunts terminated by 31 microm wide, 5 microm deep micropores. Theoretical values for the concentration factor determined by a flow circuit model in conjunction with finite volume modeling are experimentally validated. (2) Flowing macrophages are individually trapped in 20 microm wide, 6 microm deep micropores by hydrodynamic confinement. The translocation of transcription factor NF-kappaB into the nucleus upon lipopolysaccharide stimulation is imaged by fluorescence microscopy. (3) Picoliter-sized droplets are generated at a 20 microm wide, 7 microm deep micropore T-junction in an oil stream for the encapsulation of individual E. coli bacteria cells.