Name: capillary-based centrifugal microfluidic device for size-controllable formation of monodisperse microdroplets
Uploaded: 2016-02-22T15:00:00
Description: Watch this Scientific Journal Video about Capillary-based Centrifugal Microfluidic Device for Size-controllable Formation of Monodisperse Microdroplets at JoVE.com

This work was supported by the PRESTO "Design and Control of Cellular Functions" research area of the Japan Science and Technology Agency (JST), a Grant-in-Aid for Scientific Research of Innovative Areas "Molecular Robotics" (Project No. 24104002) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan, Grant-in-Aid for Young Scientists (A) (Project No. 24680033) and Scientific Research (B) (Project No. 26280097) from the Japan Society for the Promotion of Science (JSPS), and the Creative Design for Bioscience and Biotechnology course of the School of Bioscience and Biotechnology at Tokyo Tech.

1. Fabrication of a Capillary-based Microfluidic Device
<ol>
	<li>Set up of the holders 
	Note: The holder design is presented in Figure 2A.
	<ol>
		<li>Cut out each of the four discs of the holders (Figure 2A(i)-(iv)) from 2-mm-thick polyacetal plastic plate using a milling machine. Use the following dimensions for each of the four discs of the holder: (i) disc 1 diameter 8.5 mm, capillary hole (CH) diameter 1.3 mm, screw hole (SH) diameter 1.8 mm; (ii) disc 2 di...

<ol>
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Using this device, the monodisperse W/O microdroplets were generated by Plateau-Rayleigh instability of a jet-flow17. Microscopic examination did not reveal the presence of satellite droplets. In the fabrication of the device, three critical steps are essential to successfully generate monodisperse W/O microdroplets. First, to supply a straight flow of oil containing surfactant and aqueous solution, the capillary holes of four discs must be arranged in a concentric pattern. Second, the inner capillary was care...

W/O microdroplets1-5 have many important applications for the study of biochemistry and bioengineering, including protein synthesis6, protein crystallization7, emulsion PCR8,9, cell encapsulation10, and construction of artificial cell-like systems5,6. To produce W/O microdroplets for these applications, important criteria are control of size and monodispersibility of the W/O microdroplets. Microfluidic devices for making monodisperse, size-controllable W/O microdroplets11 are based on the co-flowing method12,13, flow-focusing method14,15, and the T-junction method16 in microchannels. Although these methods produce highly monodisperse W/O microdroplets, the microfabrication process requires complicated handling and specialized techniques for the preparation of microchannels, and also requires a large amount of sample solution (at least several hundred &#181;l) because of the inevitable dead volume in the syringe pumps and tubes that conduct the sample solution to the microchannels. Thus, an easy-to-use and low-dead-volume method to generate monodisperse W/O microdroplets is needed.
This paper, along with videos of experimental procedures, describes a centrifugal capillary-based axisymmetric co-flowing microfluidic device17 for generating cell-sized, monodisperse W/O microdroplets (Figure 1). This simple method achieves size monodispersity and size controllability. It requires just a tabletop mini-centrifuge and a capillary-based axisymmetric co-flowing microfluidic device fixed in a sampling microtube. Our method needs only a very small volume (0.1 &#181;l), and does not waste any significant volume of the sample.

<table border="0" cellpadding="0" cellspacing="0" width="747">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">2-mm-thick polyacetal plastic plate</td>
			<td style="width:71px;">Tool</td>
			<td style="width:119px;">Nikkyo Technos, Co., Ltd. (Japan)</td>
			<td style="width:167px;">244-6432-08</td>
			<td style="width:175px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Milling machine</td>
			<td style="width:71px;">Tool</td>
			<td style="width:119px;">Roland DG Co., Ltd. (Japan)</td>
			<td>MDX-40A</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">End Mill RSE230-0.5*2.5</td>
			<td style="width:71px;">Tool</td>
			<td style="width:119px;">NS Tool Co., Ltd. (Japan)</td>
			<td>01-00644-00501</td>
			<td></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;">M2*40 screws</td>
			<td style="width:71px;">Tool</td>
			<td style="width:119px;">Jujo Synthetic Chemistry Labo. (Japan)</td>
			<td>0001-024</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Glass Capillry Puller</td>
			<td style="width:71px;">Tool</td>
			<td style="width:119px;">Narishige (Japan)</td>
			<td>PC-10</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Microforge</td>
			<td style="width:71px;">Tool</td>
			<td style="width:119px;">Narishige (Japan)</td>
			<td>MF-900</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Inner Glass Capillary</td>
			<td style="width:71px;">Tool</td>
			<td style="width:119px;">Narishige (Japan)</td>
			<td>G-1</td>
			<td></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Outer Glass Capillary</td>
			<td style="width:71px;">Tool</td>
			<td style="width:119px;">World Precision Instruments Inc. (USA)</td>
			<td>1B200-6</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">1.5 ml Sample tube</td>
			<td style="width:71px;">Tool</td>
			<td style="width:119px;">INA OPTIKA CO.,LTD (Japan)</td>
			<td>ST-0150F</td>
			<td></td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">Hexadecane</td>
			<td style="width:71px;">Reagent</td>
			<td style="width:119px;">Wako Pure Chemical Industries Ltd. (Japan)</td>
			<td>080-03685&#160;</td>
			<td></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Sorbitan monooleate (Span 80)</td>
			<td style="width:71px;">Reagent</td>
			<td style="width:119px;">Tokyo Chemical Industry Co., Ltd. (Japan)</td>
			<td>S0060</td>
			<td></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Milli Q system</td>
			<td style="width:71px;">Reagent</td>
			<td style="width:119px;">Merck Millipore Corporation (Germany)</td>
			<td>ZRQSVP030</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Swinging-out-type Mini-centrifuge</td>
			<td style="width:71px;">Tool</td>
			<td style="width:119px;">Hitech Co., Ltd. (Japan)</td>
			<td>ATT101</td>
			<td></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Digital Microscope</td>
			<td style="width:71px;">Tool</td>
			<td style="width:119px;">KEYENCE Corporation (Japan)</td>
			<td>VHX-2001</td>
			<td></td>
		</tr>
	</tbody>
</table>

capillary-based centrifugal microfluidic device for size-controllable formation of monodisperse microdroplets

Here, we demonstrate a simple method for the rapid production of size-controllable, monodisperse, W/O microdroplets using a capillary-based centrifugal microfluidic device. W/O microdroplets have recently been used in powerful methods that enable miniaturized chemical experiments. Therefore, developing a versatile method to yield monodisperse W/O microdroplets is needed. We have developed a method for generating monodisperse W/O microdroplets based on a capillary-based centrifugal axisymmetric co-flowing microfluidic device. We succeeded in controlling the size of microdroplets by adjusting the capillary orifice. Our method requires equipment that is easier-to-use than with other microfluidic techniques, requires only a small volume (0.1-1 &#181;l) of sample solution for encapsulation, and enables the production of hundreds of thousands number of W/O microdroplets per second. We expect this method will assist biological studies that require precious biological samples by conserving the volume of the samples for rapid quantitative analysis biochemical and biological studies.

In this study, we present a simple method for the generation of cell-sized W/O microdroplets by using a capillary-based centrifugal microfluidic device (Figure 1). The microfluidic device was composed of a capillary holder (Figure 2B), two glass capillaries (inner and outer glass capillaries in Figure 3C), and a microtube containing an oil including surfactant. We injected 0.1 &#181;l of sample solution into the inner glass capillary and ...

Watch this Scientific Journal Video about Capillary-based Centrifugal Microfluidic Device for Size-controllable Formation of Monodisperse Microdroplets at JoVE.com

Capillary-based Centrifugal Microfluidic Device for Size-controllable Formation of Monodisperse Microdroplets

Here, we demonstrate a simple production method for size-controllable, monodisperse, water-in-oil (W/O) microdroplets using a capillary-based centrifugal microfluidic device. This method requires only a small sample volume and enables high-yield production. We expect this method will be useful for rapid biochemical and cellular analyses.

Here, we demonstrate a simple method for the rapid production of size-controllable, monodisperse, W/O microdroplets using a capillary-based centrifugal ...

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