Abstract

Droplet microfluidics has appealed to many interests for its capability to epitomize cells in a microscale environment and it is also a forceful technique for high-throughput single-cell epitomization. A dielectrophoretic microfluidic system imitates the oviduct of mammals with a microchannel to achieve fertilization in vitro (IVF) of an imprinting control-region (ICR) mouse. We applied a microfluidic chip and a positive dielectrophoretic (p-DEP) force to capture and to screen the sperm for the purpose of manipulating the oocyte. The p-DEP responses of the oocyte and sperm were exhibited under applied bias conditions (waveform AC 10 Vpp, 1 MHz) for trapping 1 min. The insemination concentration of sperm nearby the oocyte was increased to enhance the probability of natural fertilization through the p-DEP force trapping. A simulation tool (CFDRC-ACE+) was used to simulate and to analyze the distribution of the electric field. The DEP microfluidic devices were fabricated using poly (dimethylsiloxane) (PDMS) and ITO (indium tin oxide)-glass with electrodes. We discuss the requirement of sperm in a DEP microfluidic chip at varied concentrations to enhance the future rate of fertilization in vitro for an oligozoospermia patient. The result indicates that the rate of fertility in our device is 17.2 ± 7.5% (n = 30) at about 3000 sperms, compatible with traditional droplet-based IVF, which is 14.2 ± 7.5% (n = 28).

Highlights

  • A biochip combining microelectromechanical systems (MEMS) and biomedical technologies has the advantages of biocompatibility, high precision, and processability

  • Using the dielectrophoretic microfluidic device, the fertilization rate increased to exceed the level of DEP treatment in traditional In vitro fertilization (IVF), and more embryos developed to the blastocyst stage, with a low sperm/oocyte ratio [7]

  • To confirm that oocytes and sperm will not lyse under the applied electric field, we calculated the transmembrane voltage of oocytes and sperm as spherical cells as follows [21,26]

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Summary

Introduction

A biochip combining microelectromechanical systems (MEMS) and biomedical technologies has the advantages of biocompatibility, high precision, and processability. The droplet microfluidic technology has attracted a lot of interest because of its ability to miniaturize cells and many reagents in the microscopic environment It is an effective technique for high-throughput single-cell miniatures [1]. Using the dielectrophoretic microfluidic device, the fertilization rate increased to exceed the level of DEP treatment in traditional IVF, and more embryos developed to the blastocyst stage, with a low sperm/oocyte ratio [7]. We present a dielectrophoretic chip with ITO-glass electrodes that can serve to capture and to screen the sperm for the purpose of manipulating the oocyte of the ICR (Institute of Cancer Research) mouse, and to concentrate the sperm at the same locations for embryo formation. We tested whether a poorly conducting DEP buffer solution is usable for mouse fertilization in vitro, and we compared the rate of fertilization in vitro between a control group and an experimental group at varied concentrations of sperm

Preparation of ICR Mouse Oocytes and Sperm
AC Dielectrophoresis
Experimental Process for Standard IVF as a Control Group
Experimental Process Using the Microfluidic Chip
Findings
Conclusions

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