Design and Fabrication of All-in-One Unified Microfluidic Chip for Automation of Embryonic Cell Manipulation

Abstract

We developed a microfluidic chip for automation of cloning process based on a new protocol. The protocol is based on removal of the zona pellucida outside the chip which contributes to simplify on-chip automation of cloning. Then, the oocytes are put into the chip. The design concept of the chip is summarized as follows. (1) The oocyte is cut into two parts. (2) The divided half oocyte is sorted with and without nucleus. (3) The half oocyte without nucleus is coupled with a donor cell, and (4) they are fused by an electrical field. For the current study, the all-in-one unified microfluidic chip was designed to execute (1) cutting, (2) sorting, and (3) coupling parts continuously for this process. Basic functions of these parts as well as fusion part are verified independently. Then, all-in-one unified microfluidic chip was successfully designed and fabricated.

3D-microfluidic device to remove zona pellucida fabricated by Mask-less exposure technology

“…Recently, automation of micromanipulations on a chip is reported in order to obtain efficient, accurate and high throughput cell manipulations by using magnetic actuators [13]- [15], microfluidic forces [16] and dielectrophoretic force [17]. Figure 3 shows the swine oocyte surrounded by zona pellucida which has a role to protect oocyte during culturing.…”

“…By removing of zona pellucida at the initial stage, the oocyte can be cut easily by microfluidic force [18] with an assist of the Cytochalasin because there is no zona pellucida. Also, the donor cell can be transported to the surface of the oocyte simply by a microfluidic and dielectrophoretic force to make a coupling and fusion simply because there is no zona pellucida on the surface of oocyte [17]. Removing of zona pellucida is effective not only for the automation of cloning technique as described above, but also it is effective in the field of fertilization treatment to recover the low quality of oocyte by removing of zona pellucida in termed of the embryo development.…”

Generation of rotational flow for formation of spheroid by using microfluidics device

“…Because of these features, PDMS has been used for micro fluidic systems [7]. Researches that various actuators are embed in PDMS microchannel have been conducted, i.e.…”

Simultaneous Loading and Injection Chip to Automate Single-cell Injections for Bovine Oocytes

“…(1) Unfortunately, the technique itself cannot keep up with the growth and demand of biological research fields owing to its need for highly trained operators and its time-consuming and invasive nature. (2)(3)(4)(5) This situation has encouraged the development of various other methods of intracellular delivery, such as electroporation and the use of photothermal nanoblades, viral vector-mediated systems, and bubble injectors. (6)(7)(8)(9) Among these methods, two of the most promising methods are electroporation and the use of bubble injectors, with electroporation being the better known owing to its generally high throughput and high efficiency.…”