Copyright © 2010 by ASME Proceedings of ASME 2010 First Global Congress on NanoEngineering for Medicine and Biology NEMB2010 February 7-10, 2010 Houston, TX, USA DEVELOPMENT OF A MICROFLUIDIC INJECTION AND PERFUSION DEVICE FOR SINGLE CELL STUDY Hsiu-hung Simon Chen, Zhiquan Shu, Lei Cheng and Dayong Gao Department of Mechanical Engineering University of Washington, Seattle WA USA Corresponding author: hhchen@u.washington.edu ABSTRACT 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. Keywords: intracytoplasmic sperm injection, cryopreservation, microfluidic, cell therapy. INTRODUCTION Osmotic response of cells toward their extracellular environment plays a critical role in the fundamental study of cryobiology [1-6]. Molecules permeating through cell membranes through perfusion allow cell behaviors be monitored in the applications of drug delivery, cell therapy, and cryopreservation. On the other hand, direct injection of materials to cells through tiny needles has also shown its potentials in transgenic research and genetic engendering [7-9]. Various approaches have been utilized to quantify the cell membrane transport properties, reviewed by McGrath [10], and to manipulate single cells [11-14]. On the other hand, intracytoplasmic sperm injection (ICSI) technique has performed its successful way in in vitro fertilization (IVF) and other transgenic experiments. In order to accelerate the improvement of gene engineering, as well as lower the cost of performing ICSI related experiment (over USD100,000 for equipment itself), a cost-effective and easy-to-use system needs to be developed. In this study, a micro- nano-fluidic system that allows perfusion and injection in nano-liter scale has been realized and fabricated by soft lithographic methods. A single cell in the microfluidic system will be able to be trapped on site and then either be perfused by various solutions or injected with plain solutions or solutions with genetic materials. Before, during and after the cell is manipulated, cell behavior towards its extracullular environment will be monitored under the microscope. Our ongoing study will demonstrate that the micro- nano-fluidic system should allows us to: 1) confine cells at a in a channel; 2) deliver drugs by perfusing the cell; 3) monitor osmotic behaviors of the cell when replacing its extracellular environment; and 4) perform ICSI with sperms or genetic materials. EXPERIMENTAL METHODS Proceedings of ASME 2010 First Global Congress on NanoEngineering for Medicine and Biology NEMB2010 February 7-10, 2010 Houston, TX, USA NEMB2010-13 1 Copyright © 2010 by ASME Downloaded From: http://proceedings.asmedigitalcollection.asme.org/ on 08/25/2014 Terms of Use: http://asme.org/terms