KR101889297B1

KR101889297B1 - Apparatus for 3-dimension patterning of particles in a hydrogel and method thereof

Info

Publication number: KR101889297B1
Application number: KR1020160022730A
Authority: KR
Inventors: 이형석; 강병준; 류찬열
Original assignee: 연세대학교 산학협력단
Priority date: 2016-02-25
Filing date: 2016-02-25
Publication date: 2018-08-20
Also published as: KR20170100314A

Abstract

The present invention relates to a particle separating and aligning apparatus for separating and aligning two or more kinds of particles mixed in a fluid, comprising a fluid receiving channel for receiving a fluid containing two or more kinds of particles, And a second standing wave adding means for adding a second standing wave to the fluid receiving channel, wherein the second standing wave has a frequency corresponding to a multiple of the first standing wave . Still further, it may further comprise means for displacing the phase of the first standing wave.

Description

[0001] APPARATUS FOR 3-DIMENSION PATTERNING PARTICLES IN A HYDROGEL AND METHOD THEREOF [0002]

The present invention relates to an apparatus and a method for patterning particles in a hydrogel, and more particularly, to a method and apparatus for patterning fine particles such as cells mixed in a hydrogel in a state of fluidity before gelation of the hydrogel is three- The present invention relates to an apparatus and a method for aligning an object.

A hydrogel refers to a porous material made of a water-soluble polymer material. The porous material is a material having a pore structure therein. The porous material may have mechanical strength, permeability, electroconductivity, and the like depending on factors such as porosity, pore size, distribution, Properties have characteristics that change. These characteristics are applied to various applications such as filters, electrodes, gas sensors, gas seperators, scaffolds for wet tissue implantation, wet dressings, and mask pack materials have.

Hydrogels have fluidity before they are gelated. From an application point of view, it is necessary to align the fine particles mixed in the hydrogel three-dimensionally. For example, in the case of cells, they are subjected to three-dimensional adherence in the extracellular matrix (ECM), and the arrangement of cells in the tissue is known to have a great influence on the function. For this reason, studies are underway to identify various phenomena that cells show in an aligned state. However, there are limitations in simulating the environment of three-dimensional cell binding in a hydrogel, which is best known for cell culture.

Recently, cell sorting in a three-dimensional environment has been studied. Three-dimensional cell sorting methods include dielectric electrophoresis (DEP) and lithography, Methods have been studied. First, the method using dielectrophoresis (DEP) and lithography is a method of aligning the cells inside the hydrogel by dielectrophoresis and then fixing the position of the cells by curing the hydrogel using the lithography method. However, this approach has the problem of reducing cell viability by 5%. In addition, since the lithography method is used, there is a restriction that can be used only in hydrogels which are synthesized by light.

As one of the most powerful methods for aligning particles in a fluid, a method using a surface acoustic wave (SSAW) is known.

When a surface acoustic wave in a steady state is used, fine particles or cells can be fixed at desired positions in a fluid, and particles can be moved to desired positions through phase displacement of a surface acoustic wave. Through these devices, different particles or cells can be arranged in a three-dimensional structure or stacked in a layer structure to simulate living tissue, and promotion of growth and differentiation by cell sorting is possible.

FIG. 1 shows a conventional particle aligning apparatus using a surface acoustic wave. An IDT (Interdigital Transducer) electrode 3 for attaching a surface acoustic wave is attached to two points on the substrate 1, and a droplet 9 in which fine particles are mixed is placed between the two IDT electrodes 3. Surface acoustic waves of the same frequency are generated in the two IDT electrodes 3 and surface acoustic waves of the same frequency are superimposed to generate a standing wave. Then, the standing wave is transmitted as energy to the inside of the fluid, and the fine particles 302 in the droplet 9 are aligned to positions corresponding to the nodes of the standing waves as shown in FIG. In such a configuration, it is possible to adjust the position where the microparticles are aligned by changing the phase of the surface acoustic wave or changing the frequency.

However, the above-mentioned prior art is implemented only in a liquid such as water, and is not used for fine particle sorting such as a cell in a hydrogel. Only alignment in a two-dimensional plane is considered by standing wave, There is no technical perception of the use of dimensional alignment (alignment in the plane direction as well as height in the height direction).

U.S. Patent Publication No. 8,998,483 (Apr. U.S. Patent Publication No. 7,601,267 (Oct. 13, 2009)

Disclosure of Invention Technical Problem [8] Accordingly, the present invention has been made to solve the above-mentioned problems of the prior art, and it is an object of the present invention to provide a hydrogel capable of remarkably increasing cell survival probability And to provide a patterning apparatus and method.

An apparatus for three-dimensionally patterning particles in a hydrogel, comprising: a hydrogel receiving means for receiving a hydrogel containing particles; And a standing wave adding means for adding a horizontal standing wave to the hydrogel accommodated in the hydrogel receiving means, wherein particles in the hydrogel are arranged at positions corresponding to the nodes of the standing wave in the horizontal direction, And the vertical direction acoustic wave generated in the hydrogel is reflected by the horizontal standing wave to be superimposed on the vertical direction acoustic wave, And a lid is provided on the upper side of the hydrogel receiving means so that vertical standing waves are generated. Here, the particles may be cells or proteins, and the shape of the particles is not particularly limited, and the present invention can be applied to any type of particles.

In one embodiment of the present invention, the standing wave adding means may be a surface acoustic wave generating means or an ultrasonic transducer. Preferably, the standing wave adding means includes a substrate and a pair of IDT electrodes formed on the substrate And the surface acoustic wave generating means is a surface acoustic wave generating means.

Further, the hydrogel receiving means is disposed between the pair of IDT electrodes, and a coupling liquid may be interposed between the hydrogel receiving means and the substrate. .

The hydrogel receiving means includes a block that encloses the side surface of the chamber, which is a space where the hydrogel is located, and a lid that covers the upper surface of the chamber. And the vertical alignment interval of the particles in the hydrogel,

(Where V _liquid is the propagation velocity of the wave inside the hydrogel, V _SAW is the propagation velocity of the surface acoustic wave on the substrate, and? _SAW is the wavelength of the surface acoustic wave).

Further, the present invention provides a method for three-dimensionally patterning particles in a hydrogel, comprising: determining a particle alignment interval in the horizontal and vertical directions within the hydrogel; Determining an interval of the IDT electrodes from the horizontal alignment interval; Determining a material of the hydrogel and the substrate from the vertical alignment spacing; Disposing a hydrogel in the hydrogel receiving means; And applying AC power to the IDT electrode.

It is possible not only to align the particles such as proteins or cells in the hydrogel three-dimensionally but also to adjust the vertical alignment interval as well as the horizontal alignment interval according to need.

1 is a perspective view of a particle aligning apparatus using a surface acoustic wave device according to the prior art.
Fig. 2 shows a state in which particles are aligned by surface acoustic waves having normal fines.
3 is a side view of an apparatus for patterning particles in a hydrogel according to an embodiment of the present invention.
4 is a plan view of an apparatus for patterning particles in a hydrogel according to an embodiment of the present invention.
5 is a side view of an apparatus for patterning particles in a hydrogel according to another embodiment of the present invention.
6 illustrates a structure of an IDT electrode according to an embodiment of the present invention.
FIG. 7 shows a state in which particles are three-dimensionally patterned in the hydrogel.
8 shows a state in which a standing wave in the horizontal direction and a standing wave in the vertical direction are formed by the IDT electrode.
9 is a flowchart illustrating a method of patterning a hydrogel particle according to an embodiment of the present invention.

FIG. 3 is a side view of a particle patterning apparatus 100 for three-dimensionally aligning particles in a hydrogel according to a first embodiment of the present invention, FIG. 4 is a plan view of a particle patterning apparatus 100 according to the above- to be. Here, the meaning of three-dimensionally aligning the particles in the hydrazine gel means that the particles are aligned at regular intervals in the height (vertical) direction, apart from the arrangement of the particles in a line in a plane .

The particle patterning apparatus 100 in the hydrogel includes a substrate 110, a hydrogel 200 receiving means 120 formed on the substrate, and a substrate holder 120 symmetrically disposed on the substrate And IDT electrodes 130L and 130R formed on the substrate 110. FIG.

The substrate 110 can be mainly made of lithium niobate (LiNbO ₃ ), quartz (Quartz), or lithium tantalite (LiTaO ₃ ), but the material is not particularly limited if it is a material capable of generating surface acoustic waves.

On the substrate 110, a hydrogel receiving means 120 capable of receiving a hydrogel is formed. 3, a structure of the hydrogel receiving means 120 includes a first block 123 formed on a substrate 110 and a first cover 124 formed on an upper end of the first block 123 do. A second block 125 is formed on the upper end of the first cover 124 and a second cover 121 is formed on the upper end of the second block 125.

Here, the first block 123 and the second block 125 may be made of PDMS (polydimethylsiloxane), but any material that can serve as a wall structure for forming a space capable of accommodating a substance such as liquid or hydrogel It is available. As the first cover 122 and the second cover 121, a glass substrate may be preferably used. However, it is also possible to use any material such as a liquid or a hydrogel and a material that can serve as a structure capable of blocking the outside. It is possible.

The first block 123 is interposed between the substrate 110 and the first cover 122 to serve as a column for supporting the first cover 124. The first block 123 has an inner space The coupling liquid 124 may be filled. The coupling liquid 124 may be water (distilled water), ethylene glycol, glycerol, mercury, or the like, but it is preferable that the coupling energy of the wave energy of the surface acoustic wave transmitted through the substrate 110 Anything that can deliver liquid is possible.

The second block 125 is interposed between the first cover 122 and the second cover 121 to serve as a column for supporting the second cover 121. The second block 125 is disposed between the hydrogel 200 ) Serves as a wall structure defining the space in which it is received.

Since the coupling liquid 124 is interposed between the space in which the hydrogel 200 is accommodated and the substrate 110, the structure for accommodating the hydrogel 200 after the patterning process of the hydrogel 200 is separated from the substrate 110 So that it can be easily separated. In the embodiment shown in FIG. 3, the convenience of the process is improved by using the coupling liquid. However, as shown in FIG. 5, when the hydrogel 200 is in contact with the substrate 110 without interposing the coupling liquid The three-dimensional patterning of the hydrogel 200 can be performed.

Standing wave generating means may be formed on both sides of the hydrogel receiving means with the upper hydrogel receiving means 120 therebetween. In the embodiment of the present invention, the IDT electrodes 130L and 130R are used as the standing wave generating means, but it is also possible to use a stimulating means capable of generating a standing wave by generating standing waves, for example, an ultrasonic transducer using a piezoelectric element It is possible.

The IDT electrode is formed by attaching a piezoelectric material to the substrate 110. When an AC power source is applied to the IDT electrodes 130R and 130L, the IDT electrode generates surface acoustic waves on the substrate 110. [ The pair of IDT electrodes 130R and 130L disposed on both sides of the hydrogel receiving means 120 generate surface acoustic waves of the same frequency. Surface acoustic waves of the same frequency are superimposed on each other, Wave.

A standing wave is a concept that contrasts with a progressive wave, which travels in an arbitrary direction. It refers to a wave whose node of vibration is fixed at a certain position. When waves of the same amplitude and vibration move in opposite directions It is caused by the synthesis of waves.

FIG. 6 shows a basic structure of an IDT electrode. The IDT electrode is formed by forming a piezoelectric material on a substrate by a method such as evaporation. The IDT electrodes 130R and 130L are formed in such a manner that the upper electrode 131 and the lower electrode 132 are engaged with each other and the wavelength or frequency of the surface acoustic wave generated by adjusting the width and the interval of the electrode branches can be adjusted The width of one electrode branch and the interval between the electrode branch and the adjacent electrode branch are determined to be 1/4 of the wavelength (? SAW / 4)).

For convenience of explanation, the wavelength of the surface acoustic wave generated by each of the IDT electrodes 130R and 130L is set to "? _SAW " It should be noted.

As described above, the standing wave is a wave in which a nodal point of vibration is not moved but is maintained in a certain region, and the upper node is formed at intervals of? _SAW / 2.

The standing wave is formed along the surface of the substrate 110. When the coupling fluid 124 is contacted with the coupling fluid 124, a standing wave is propagated in the vertical direction, and a standing wave is transmitted to the hydrogel 200 through the first cover 122.

8, the standing wave 410 in the horizontal direction is formed by the IDT electrode and the upper standing wave is transmitted to the hydrogel 200. In the hydrogel 200, A standing wave 420 is generated in the direction of the arrow.

When a standing wave is formed in the hydrogel, the particles have a property of being aligned in the vicinity of the node of the standing wave, so that the particles in the hydrogel 200 are aligned not only in the horizontal direction but also in the height Three-dimensional alignment of particles is implemented.

At this time, the horizontal standing wave applied to the hydrogel 200 is the same as the standing waves generated by the IDT electrodes 130R and 130L, but the vertical standing waves formed in the hydrogel 200 are the IDT electrodes 130R and 130L ) Electrode having a wavelength different from that of the standing wave generated by the electrode, and can be derived specifically by the following equation.

Here,? _SAW is a wavelength of a surface acoustic wave generated from each of the IDT electrodes 130R and 130L. When a standing wave is formed, a node of a standing wave is formed at a half wavelength length interval of the surface acoustic wave. Since the particles are collected near the nodes of the standing waves, the horizontal interval (d _horizontal ) in which the particles are aligned is determined by the half wavelength of the surface acoustic wave (see Equation 1).

When the upper standing wave reaches the hydrogel or fluid, a standing wave in the vertical direction is generated as shown in FIG. 8, and the particles are aligned at regular intervals in the vertical direction by the standing wave in the vertical direction. The vertical spacing (d- _vertical ) in which the particles are aligned is equal to the half-wave length of the vertical standing wave, which is the velocity of wave propagation (V _liquid ; sound velocity) within the hydrogel, the surface acoustic wave propagation velocity (V _SAW ) , And the wavelength ( _SAW ) of the surface acoustic wave.

Referring to FIG. 9, first, horizontal and vertical alignment intervals of particles to be aligned in the hydrogel are determined (S100), and the interval of the horizontal standing wave and the wavelength of the surface acoustic wave to be applied from the horizontal alignment interval are determined . When the wavelength of the surface acoustic wave is determined, the interval of the IDT electrodes is determined as shown in FIG. 6 (S200). The vertical alignment interval is determined by the wave propagation velocity (V _liquid ; sound velocity) inside the hydrogel, the surface acoustic wave propagation velocity (V _SAW ) on the substrate, and the wavelength of the surface acoustic wave (? _SAW ) Since the wavelength of the surface acoustic wave is determined, the physical properties of the hydrogel and the physical properties of the substrate that affect the wave propagation speed can be determined (S300). When the hydrogel is disposed in the hydrogel receiving means of the device obtained through the above-described designing process (S400) and AC power is applied to the IDT electrode (S500), the particles in the hydrogel not only in the horizontal direction but also in the vertical direction So that they can be aligned at desired intervals.

As a reference material for the hydrogel used in the present invention, a variety of synthetic polymer materials and natural polymer materials can be used. Representative synthetic polymer materials include polyacrylamide (PAAM), hyaluronic acid (HA) ) Or Hyaluronic Acid Catechol (HACA), polyethylene glycol (PED), polyvinyl alcohol, polyethylene oxide and the like. Natural polymer materials include collagen and polysaccharides Chitosan, alginate, agarose), and the like can be used.

100: Particle patterning device in hydrogel 110:
120: hydrogel receiving means 121: first cover
122: second cover 123: first block
124: coupling liquid 125: second block
130L, 130R: IDT electrode 131: upper electrode
132: lower electrode 200: hydrogel

Claims

An apparatus for three-dimensionally patterning particles in a hydrogel,
A hydrogel receiving means for receiving a hydrogel containing particles; And
And a standing wave adding means for adding a horizontal standing wave to the hydrogel contained in the hydrogel receiving means,
The particles are aligned in a position corresponding to a node of a standing wave in the horizontal direction and aligned in a position corresponding to a node of a vertical standing wave generated in the hydrogel by the standing wave in the horizontal direction,
And a lid is provided on the upper side of the hydrogel receiving means to reflect the vertical acoustic waves generated in the hydrogel by the horizontal standing waves and to generate the vertical standing waves through superimposition with the vertical acoustic waves. An apparatus for three-dimensionally patterning particles in a hydrogel

The apparatus for three-dimensionally patterning particles in a hydrogel according to claim 1, wherein the stationary wave applying means is a surface acoustic wave generating means or an ultrasonic transducer.

The apparatus for three-dimensionally patterning particles in a hydrogel according to claim 2, wherein the standing wave adding means is a surface acoustic wave generating means including a substrate and a pair of IDT electrodes formed on the substrate.

4. The apparatus for three-dimensionally patterning particles in a hydrogel according to claim 3, wherein the hydrogel receiving means is provided between the pair of IDT electrodes.

The apparatus for three-dimensionally patterning particles in a hydrogel according to claim 3, wherein a coupling liquid is interposed between the hydrogel receiving means and the substrate.

4. The apparatus of claim 3, wherein the hydrogel receiving means comprises a block surrounding a side of the chamber which is a space in which the hydrogel is located.

The method according to claim 3, wherein the vertical alignment interval of the particles in the hydrogel

Wherein the particle size of the particles in the hydrogel is determined by the particle size distribution.
(Where V _liquid is the propagation velocity of the wave inside the hydrogel, V _SAW is the propagation velocity of the surface acoustic wave on the substrate, and? _SAW is the wavelength of the surface acoustic wave)

The apparatus for three-dimensionally patterning particles in a hydrogel according to claim 1, wherein the particles are proteins or cells.

A method for patterning using a device for three-dimensionally patterning particles in a hydrogel according to claim 3,
Determining a particle alignment interval in the horizontal and vertical directions within the hydrogel;
Determining an interval of the IDT electrodes from the horizontal alignment interval;
Determining a material of the hydrogel and the substrate from the vertical alignment spacing;
Disposing a hydrogel in the hydrogel receiving means; And
And applying AC power to the IDT electrode. A method for three-dimensionally patterning particles in a hydrogel.