The cell membrane, composed primarily of proteins and lipids, is a selectively permeable lipid bilayer in the scale of 10 nm or so. Molecules permeating through cell membranes play critical roles in the applications of drug delivery, cell therapy, and cryopreservation. Cryopreservation and banking of cells, such as umbilical cord bloods, female eggs, etc., are critical to facilitate practical and effective in vitro fertilization (IVF). The determination of molecule transport properties of cells, such as water and cryoprotectants (CPAs), is indispensable for developing optimal conditions for cryopreserving them. On the other hand, injection of material of interests, such as sperms and DNA segments, to female eggs or blastocysts, so-called intracytoplasmic sperm injection (ICSI) technique, are playing important roles on IVF and advanced gene knock-out. In this study, a novel micro-nano-fluidic system that allows perfusion and injection in nano-liter scale has been developed and fabricated by soft lithographic methods. A single cell in the microfluidic system is able to be trapped on site and then either be perfused by various solutions or injected with plain solutions or solutions with genetic materials. Our ongoing study will demonstrate that the micro-nano-fluidic system allows us to: 1) confine cells in a channel; 2) deliver drugs by perfusing the cell; 3) monitor osmotic behaviors of the cell by replacing its extracellular environment; and 4) perform ICSI with sperms or genetic materials.

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