KR102057788B1 - Continuously sedimentation-based microfluidic separation and cell culture system and method using hydraulic jump phenomenon - Google Patents

Abstract

The present invention relates to a continuous sedimentation-based microfluidic separation and culture system using hydraulic leap phenomenon and to a method thereof. The continuous sedimentation-based microfluidic separation and culture system using hydraulic leap phenomenon according to an embodiment comprises: an inlet port for inducing a sample; an outlet port through which the sample is discharged; and a capture space portion for capturing at least one target particle included in the sample disposed between the inlet port and the outlet port so as to have height higher than that of the inlet port and the outlet port, and flowing from the inlet port toward the outlet port using the hydraulic leap phenomenon.

Description

CONTINUOUSLY SEDIMENTATION-BASED MICROFLUIDIC SEPARATION AND CELL CULTURE SYSTEM AND METHOD USING HYDRAULIC JUMP PHENOMENON}

The description below is a technique for separating and culturing microscale particles (hereinafter, the particles include cells), and more specifically, selectively separating at least one target particle using a hydraulic jump phenomenon. The present invention relates to a system and a method for continuously separating and culturing by culturing at least one target particle separated and captured.

Techniques for separating target particles using hydraulic leap phenomena (Appl. Phys. Lett. 97, 154101 (2010); doi: 10.1063 / 1.3479052, Biomicrofluidics 8, 064108 (2014); doi: 10.1063 / 1.4902906 , Hamidreza Shirinkami et al lab on a chip, 2018 (submitted) Large-scale microchip for size-selective particle and cell sorting based on hydraulic jump phenomenon.

One embodiment utilizes a hydraulic jump phenomenon to selectively separate at least one particle (hereinafter referred to as at least one target particle) of desired physical properties (size and density) among a plurality of particles included in a sample. And systems and methods for capturing.

More specifically, one embodiment provides a system and method for selectively capturing at least one target particle by placing at least one capture medium at the dropping point of the at least one target particle in view of the settling velocity of the at least one target particle. To provide.

At this time, one embodiment provides a system and method for continuously separating and culturing by culturing at least one target particle captured using at least one capture medium.

In addition, one embodiment provides a system and method for selectively separating and capturing at least one target particle by determining and adjusting the length of the capture space based on the settling velocity of the at least one target particle.

In addition, one embodiment provides a system and method for maximizing throughput for capturing at least one target particle by determining and adjusting the width of the capture space.

According to one embodiment, a continuous sedimentation-based microfluidic separation and culture system using a hydraulic leap phenomenon, the sample inlet; An outlet through which the sample is discharged; And at least one target particle included in the sample disposed between the inlet and the outlet so as to have a height higher than that of the inlet and the outlet, and flowing from the inlet toward the outlet. And a capture space portion for capturing.

According to one aspect, the capture space portion, the height of the capture space portion is sharply higher than the height of the inlet port can be captured by seizing the at least one target particles as the movement speed of the sample is lowered in the capture space portion. .

According to another aspect, the capture space portion, as the particles have different falling points and settling speed based on the size or density of each of the particles included in the sample, the at least one target particle of the particles Can be selectively captured.

According to another aspect, the capture space portion, at least one of capturing and culturing the at least one target particle while being disposed at the dropping point of the at least one target particle in consideration of the settling velocity of the at least one target particle Capture media.

According to another aspect, the continuous sedimentation-based microfluidic separation and culture system may further include an optical sensor for observing at least one target particle cultured in the at least one capture batch.

According to another aspect, the length of the capture space portion may be determined based on the sedimentation velocity of the at least one target particle.

According to another aspect, the width of the capture space portion may be determined based on the throughput of capturing the at least one target particle.

According to one embodiment, the continuous sedimentation-based microfluidic separation and culture method using a hydraulic leap phenomenon, the step of introducing a sample through the inlet; At least one target particle included in the sample flowing from the inlet toward the outlet through a capture space formed between the inlet and the outlet through which the sample is discharged, and having a height higher than that of the inlet and the outlet. Capturing using a hydraulic jump phenomenon; And discharging the sample from which the at least one target particle is captured through the discharge port.

According to one aspect, the step of capturing, by seizing and capturing the at least one target particle as the height of the capture space is sharply higher than the height of the inlet port so that the moving speed of the sample is lowered in the capture space portion Can be.

According to another aspect, the capturing may include the at least one target of the particles as the particles have different drop points and settling speeds based on the size or density of each of the particles included in the sample. Selectively capturing the particles.

According to another aspect, the capturing may include at least one target particle through at least one capture medium disposed at a dropping point of the at least one target particle in consideration of the settling velocity of the at least one target particle. It may include the step of capturing the culture.

According to one embodiment, a continuous sedimentation-based microfluidic separation and culture system using a hydraulic leap phenomenon, the sample inlet; An outlet through which the sample is discharged; And a plurality of capture space portions disposed in series or in parallel between the inlet and the outlet, wherein each of the plurality of capture spaces is formed to have a height higher than that of the inlet and the outlet, and flows from the inlet to the outlet. Different target particles contained in the sample are captured using a hydraulic jump phenomenon.

According to one aspect, each of the plurality of capture spaces, the height of each of the plurality of capture spaces is sharply higher than the height of the inlet port so that the movement speed of the sample is lowered in each of the plurality of capture spaces Other target particles can be sedimented and captured.

According to another aspect, each of the plurality of capture spaces, the particles having a different drop point and the settling speed based on the size or density of each of the particles included in the sample, the said one of the particles Other target particles can be selectively captured.

According to another aspect, each of the plurality of capture spaces, capture medium for capturing and incubating the different target particles while being disposed at the falling point of the different target particles in consideration of the settling speed of the different target particles It may include.

According to another aspect, the continuous sedimentation-based microfluidic separation and culture system may further include a plurality of optical sensors provided corresponding to each of the plurality of capture spaces to observe the target particles cultured in the capture batch. have.

According to another aspect, each of the plurality of capture spaces may be formed in different lengths based on the settling velocity of the different target particles.

According to one embodiment, the continuous sedimentation-based microfluidic separation and culture system using a hydraulic leap phenomenon, the first inlet for the first sample is introduced; A first outlet through which the first sample is discharged; A second inlet through which a second sample is introduced; A second outlet through which the second sample is discharged; And a height higher than that of the first inlet, the second inlet, the first outlet, and the second outlet, disposed between the first inlet and the first outlet and between the second inlet and the second outlet. And at least one first target particle included in the first sample and flowing from the second inlet toward the second outlet based on a hydraulic leap phenomenon. A capture space portion for capturing each of the at least one second target particle included in the second sample is included.

One embodiment can provide a system and method for using hydrodynamic leap to selectively separate and capture at least one target particle of desired physical properties (size and density) among a plurality of particles included in a sample. .

More specifically, one embodiment provides a system and method for selectively capturing at least one target particle by placing at least one capture medium at the dropping point of the at least one target particle in view of the settling velocity of the at least one target particle. Can be provided.

At this time, one embodiment may provide a system and method for continuously separating and culturing by culturing the captured at least one target particle using at least one capture medium.

In addition, one embodiment may provide a system and method for selectively separating and capturing at least one target particle by determining and adjusting the length of the capture space based on the settling velocity of the at least one target particle.

In addition, one embodiment may provide a system and method for maximizing throughput for capturing at least one target particle by determining and adjusting the width of the capture space.

1A-1C illustrate a continuous sedimentation based microfluidic separation and culture system according to one embodiment.
2A-2B illustrate a continuous sedimentation based microfluidic separation and culture system with at least one capture medium added in accordance with one embodiment.
3 is a flow chart illustrating a method of continuous sedimentation based microfluidic separation and culture according to one embodiment.
4 is a diagram illustrating a continuous sedimentation-based microfluidic separation and culture system in which a plurality of capture spaces are connected in series, according to one embodiment.
5 is a diagram illustrating a continuous sedimentation-based microfluidic separation and culture system in which a plurality of capture spaces are connected in parallel, according to one embodiment.
6A-6B illustrate a continuous sedimentation based microfluidic separation and culture system according to another embodiment.

Hereinafter, exemplary embodiments of the present invention will be described in detail with reference to the accompanying drawings. However, the present invention is not limited or limited by the embodiments. Also, like reference numerals in the drawings denote like elements.

Also, terms used in the present specification are terms used to properly express preferred embodiments of the present invention, which may vary depending on the intention of a viewer, an operator, or customs in the field to which the present invention belongs. Therefore, the definitions of the terms should be made based on the contents throughout the specification. For example, in this specification, the singular also includes the plural unless specifically stated otherwise in the text. Also, as used herein, “comprises” and / or “comprising” refers to one or more other components, steps, operations and / or elements. It does not exclude the presence or addition of devices.

In addition, it should be understood that various embodiments of the present invention are different from one another, but need not be mutually exclusive. For example, specific shapes, structures, and characteristics described herein may be implemented in other embodiments without departing from the spirit and scope of the invention with respect to one embodiment. In addition, it should be understood that the position, arrangement, or configuration of individual components in each of the exemplary embodiments presented may be changed without departing from the spirit and scope of the present invention.

The target particles described below are particles having desired physical properties (size and density) among the plurality of particles included in the sample, and mean particles to be separated and captured.

1A-1C illustrate a continuous sedimentation based microfluidic separation and culture system according to one embodiment. In more detail, Figure 1a is a top view showing a continuous sedimentation-based microfluidic separation and culture system according to an embodiment, Figure 1b is a cross-sectional view showing a continuous sedimentation-based microfluidic separation and culture system shown in Figure 1a, Figure 1c is a view for explaining a hydraulic leap phenomenon used by the continuous sedimentation-based microfluidic separation and culture system according to an embodiment.

1A to 1C, a continuous sedimentation-based microfluidic separation and culture system (hereinafter, referred to as a system) 100 according to an embodiment includes an inlet 110 through which a sample is introduced and an outlet through which a sample is discharged ( 120 and a capture space 130 disposed between the inlet 110 and the outlet 120.

The capture space 130 is formed to have a height higher than that of the inlet 110 and the outlet 120, so that at least one target particle 140 included in the sample flowing from the inlet 110 toward the outlet 120 is included. Capture using hydraulic leap phenomena.

Specifically, as shown in FIG. 1C, the sample flowing at a very high speed from the inlet 110 rapidly decreases in movement speed due to a rapid height change (H = h / h ₀ ) when entering the capture space 130. (Where h represents the height of the capture space 1300 and h ₀ represents the height of the inlet 110 and outlet 120), ie for the particles 140, 150 included in the sample. In the inlet 110, the drag force toward the outlet 120 acts more than gravity, and in the capture space 130, the gravity acts more than the drag toward the outlet 120. Particles (140, 150) included in the sample is settled in the capture space 130, bar system according to an embodiment the height of the capture space 130 is greater than the height of the inlet 110 dramatically increased the speed of the sample V _T decreases in (Traveling speed of velocity in the longitudinal direction) are trapped space 130 At least one target particle 140 using the sedimentation due to the hydraulic jump phenomenon separating at least one target particle 140 from the sample according to and can be trapped in the trapping recess 130.

At this time, when the initial velocity of the sample flowing into the inlet 110 is V ₀ , the average moving velocity is calculated as V ₀ * H ⁻¹ , and the sedimentation velocity V _S is each of the particles 140 and 150 included in the sample. It has a relation proportional to the size and density of and inversely proportional to the viscosity of the sample. Thus, as the size and density of each of the particles 140 and 150 are different, the settling velocity and the falling point of each of the particles 140 and 150 are also different. For example, the settling speed of the particles 140 having a larger size and density may be faster than the settling speed of the particles 150 having a smaller size and density, and the dropping points of the particles 140 having a larger size and density may have a larger size and density. It may be closer from the inlet 110 on the capture space 130 than the drop point of the small particles 150. Therefore, the system 100 according to an exemplary embodiment may consider the settling velocity and the dropping point of each of the particles 140 and 150 included in the sample, thereby providing at least one target particle of the plurality of particles 140 and 150. Only 140 may be selectively captured and separated.

More specifically, the settling time t _S at which each of the particles 140 and 150 included in the sample is settled may be calculated as in Equation 1 below, and the capture space of each of the particles 140 and 150 included in the sample may be calculated. The pass time t _T of the unit 130 may be calculated as in Equation 2.

<Equation 1>

h/2*V_S t _S

h / 2 * V _S

<Equation 2>

l*H/V₀ t _T

l * H / V ₀

Accordingly, the system 100 according to an embodiment may determine the length l of the capture space 130 in consideration of the settling velocity of at least one target particle 140 to be separated and captured among the particles 140 and 150. By determining and adjusting, only at least one target particle 140 can be selectively captured and separated. For example, the system 100 may include a capture space such that the settling time according to the settling velocity of the at least one target particle 140 satisfies a condition t _S <t _T less than the passage time of the at least one target particle. The length of the 130 may be determined and adjusted to separate and capture the at least one target particle 140 in the capture space 130.

At this time, the system 100 determines and adjusts the length of the capture space 130 in consideration of the settling velocity of the at least one target particle 140, and at least one of the particles 140 and 150 included in the sample. It is also possible to consider the settling velocity of the remaining particles 150 except for one target particle 140. For example, the system 100 may satisfy the condition t _S <t _T where the settling time according to the settling speed of the at least one target particle 140 is less than the passage time, while at the same time each of the remaining particles 150 The length of the capture space 130 is determined and adjusted so that the settling time according to the settling velocity satisfies a condition (t _S > t _T ) greater than the passage time, so that the at least one target particle 140 is captured. While separating and capturing in 130, the remaining particles 150 may be discharged through the outlet 120.

In addition, the system 100 according to an exemplary embodiment may include a length of the capture space 130 having a length of a drop point of at least one target particle 140 to be separated and captured among the particles 140 and 150. By determining and adjusting l , only at least one target particle 140 can be selectively captured and separated. In particular, the system 100 may further include at least one capture medium (not shown) for selectively capturing the at least one target particle 140. Detailed description thereof will be described with reference to FIGS. 2A to 2B.

Although not shown in the drawings, if the plurality of target particles are to be separated and captured, the system 100 determines the length of the capture space 130 in consideration of the settling velocity and the falling point of each of the plurality of target particles. And can be adjusted. For example, the system 100 satisfies the condition that the settling time according to the settling speed of each of the plurality of target particles is less than the passage time of each of the plurality of target particles, and includes a drop point of each of the plurality of target particles. The length of the capture space 130 may be determined and adjusted to the length to be.

In consideration of the settling velocity and the falling point of the plurality of target particles, if the capture space 130 cannot be captured and separated from the plurality of target particles only by adjusting the length of the capture space 130, the system 100 may include one capture space. It may be implemented to include a plurality of capture space portions other than the portion 130. Detailed description thereof will be described with reference to FIGS. 4 to 5.

Here, the throughput of capturing at least one target particle 140 may be improved when the average moving speed of the sample is increased, and at least one target particle 140 is reduced when the velocity of the sample in the capture space 130 is reduced. The capture rate to capture the bar can be improved, the system 100 according to an embodiment increases the speed of the sample at the inlet 110 to increase the average moving speed of the sample at the same time in the capture space 130 The sample rate can be reduced to improve both throughput and capture rate. Specifically, the system 100 according to an embodiment adaptively determines and adjusts the volume of the capture space 130 and the volume of the inlet 110 associated with the average moving speed of the sample, thereby providing at least one target particle ( It is possible to improve the throughput and capture rate to capture. For example, the system 100 reduces the volume of the inlet 110 to increase the speed of the sample at the inlet 110, and increases the volume of the capture space 130 to increase the volume of the sample at the capture space 130. By reducing the speed of the substrate, the throughput and the capture rate of capturing the at least one target particle 140 may be improved. At this time, since each of the volume of the capture space 130 and the volume of the inlet 110 is related to the width of the capture space 130 and the width of the inlet 110, the system 100 is at least one target particle ( The width of the capture space 130 and the width of the inlet 110 may be determined and adjusted based on the throughput and the capture rate for capturing 140.

The system 100 according to the exemplary embodiment may not only improve the throughput of capturing the at least one target particle 140, but also use platelet samples by widely determining and adjusting the width of the capture space 130. When used in the platelet functional test, since the shear stress is lowered in the capture space 130, the effect of capturing plasma without activating platelets can be achieved.

The at least one target particle 140 thus separated and captured may be observed by an optical sensor (not shown) further included in the system 100. Hereinafter, the at least one target particle 140 separated and captured is described as being observed by an optical sensor based on an optical method, but various sensors for observing and monitoring through an electrical method or a chemical method are not limited thereto. It can be used instead of the optical sensor.

2A-2B illustrate a continuous sedimentation based microfluidic separation and culture system with at least one capture medium added in accordance with one embodiment. More specifically, FIG. 2A is a cross-sectional view showing a state prior to culture of at least one target particle captured in a continuous sedimentation-based microfluidic separation and culture system to which at least one capture medium is added, and FIG. 2B shows at least one capture A cross-sectional view showing the state after at least one target particle captured in the continuous sedimentation-based microfluidic separation and culture system to which the medium is added is cultured.

2A to 2B, a continuous sedimentation-based microfluidic separation and culture system (hereinafter referred to as a system) 200 according to one embodiment may include at least one of the settling speeds of at least one target particle 220. By further including at least one capture medium 210 disposed at the dropping point of one target particle 220, it is possible to selectively capture at least one target particle 220.

In particular, the system 200 further includes at least one capture medium 210, thereby culturing at least one target particle 220 captured as described above with reference to FIGS. 1A-1C to continuously separate and culture. Can be done. In this case, the capture space 230 may be further provided with a light source 240 for culturing the at least one target particle 220 in the at least one capture medium 210.

For example, the size of particles of lung cancer CTC is 12.5um to 30.6um, which is much larger than 10um white blood cells and 5um red blood cells, but is present in very low concentrations in the blood. Has very difficult limitations to separate the CTCs to be observed. Therefore, in order to overcome the limitation, the system utilizing the existing size-based separation method increases the number of incoming CTCs to increase the number of captured CTCs or observes a small number of CTCs with a high performance optical sensor. It has been improved to apply a method to support the, but all of the methods have the disadvantage that additional costs are consumed.

On the other hand, the system 200 according to an embodiment separates a small number of CTCs in at least one capture medium 210 and then incubates the number of small numbers of CTCs by changing only the culture medium on the at least one capture medium 210. Increasing may support observation through an optical sensor (not shown) of pervasive performance without further processing of the sample (eg, fluorescence staining, antibody labeling, etc.). Thus, a system 200 including at least one capture medium 210 can minimize damage and loss of captured particles and the cost of particle observation. In addition, various environmental requirements are required depending on the type of sample (eg, a particle such as vascular endothelial cells requires a microenvironment requiring shear stress in culture), and the system 200 according to an embodiment By appropriately determining and adjusting the volume and width of the capture space 230 in which the at least one capture medium 210 is disposed, the microenvironment can be adaptively controlled to easily meet the environmental requirements of the sample. It has an effect that can be made.

Although the system 200 has been described as including one capture medium 210, the present invention is not limited thereto, and the system 200 is provided to correspond to the plurality of target particles if it is to separate and capture the plurality of target particles. It may comprise a plurality of capture media. In this case, the plurality of capture media may be disposed in consideration of the falling points of each of the plurality of target particles to be captured. For example, the first capture medium may be disposed at the dropping point of the first target particle, and the second capture medium may be disposed at the dropping point of the second target particle.

3 is a flow chart illustrating a method of continuous sedimentation based microfluidic separation and culture according to one embodiment. Hereinafter, the continuous sedimentation-based microfluidic separation and culture method according to one embodiment is performed by the continuous sedimentation-based microfluidic separation and culture system described below with reference to FIGS. 1A to 2B.

Referring to FIG. 3, the system according to an embodiment introduces a sample through an inlet at step S310.

Subsequently, the system includes at least one target included in the sample flowing from the inlet to the outlet through a capture space formed to have a height higher than the inlet and the outlet while being disposed between the inlet and the outlet through which the sample is discharged in step S320. Particles are captured using a hydraulic jump phenomenon. In step S320, the system captures the at least one target particle by using the hydraulic leap phenomenon, wherein the height of the capture space is drastically higher than the height of the inlet so that the velocity of the sample is lowered in the capture space. By sedimentation and capture of particles.

More specifically, the system utilizes the property that the particles have different drop points and settling rates based on the size or density of each of the particles included in the sample, so that at least one target particle of the particles in step S320 is used. Can be selectively captured. For example, the system can selectively capture only at least one target particle by determining and adjusting the design based on the sedimentation velocity and the dropping point of the at least one target particle to capture the length of the capture space.

In particular, the system may capture and incubate at least one target particle through at least one capture medium disposed at the dropping point of the at least one target particle in consideration of the settling velocity of the at least one target particle in step S320. .

The system then discharges the sample from which the at least one target particle has been captured through the outlet in step S330.

As such, the system may support observation of at least one target particle using the optical sensor by capturing at least one target particle in the capture space through steps S310 to S330. In particular, by performing culturing of at least one target particle captured in step S320, the system may support observation of the at least one target particle that has been cultured.

4 is a diagram illustrating a continuous sedimentation-based microfluidic separation and culture system in which a plurality of capture spaces are connected in series, according to one embodiment.

Referring to FIG. 4, a continuous sedimentation-based microfluidic separation and culture system (hereinafter, referred to as a system) 400 according to an embodiment may include a plurality of capture spaces 410 and 420 connected in series. have.

In this case, the length of each of the plurality of capture spaces 410, 420 may be determined and adjusted based on the settling velocity and the dropping point of each of the plurality of target particles to be captured by the system 400. For example, the first capture space 410 may set the sedimentation rate of the first target particles 430 to be separated and captured among the particles 430 and 440 included in the sample, and the second target particles to be passed ( By determining and adjusting the length in consideration of the settling velocity of 440, only the first target particles 430 may be selectively captured and separated, and the second capture space 420 may include particles 430, included in the sample. Since the length is determined and adjusted in consideration of the settling speed of the second target particles 440 to be separated and captured in the 440, only the second target particles 440 may be selectively captured and separated. More specifically, for example, the length of the first capture space 410 may include a condition t _1S <t _1T in which the settling time t _1S corresponding to the settling speed of the first target particle 430 is smaller than the passage time t _1T. The settling time t _2S according to the settling speed of the second target particle 440 is determined and adjusted to satisfy the condition t _2S > t _2T greater than the passage time t _2T , and the length of the second capture space 420 Is determined and adjusted so that the settling time t _2S according to the settling speed of the second target particle 440 satisfies a condition t _2S <t _2T less than the passage time t _2T , thereby making the first target particle 430 the first target particle 430. The first capture space 410 may be separated and captured, and the second target particles 440 may be separated and captured in the second capture space 420.

In this case, each of the plurality of capture spaces 410 and 420 may further include a capture medium (not shown) described above with reference to FIGS. 2A to 2B to continuously capture and culture the target particles. In addition, each of the plurality of capture spaces 410 and 420 may be further provided with an optical sensor provided to observe the captured and cultured target particles.

In addition, the system 400 instead of capturing and separating the plurality of target particles 430 and 440 into the plurality of capture spaces 410 and 420 connected in series as shown in FIG. It is also possible to capture and separate a plurality of target particles in the capture spaces. Detailed description thereof will be described with reference to FIG. 5.

5 is a diagram illustrating a continuous sedimentation-based microfluidic separation and culture system in which a plurality of capture spaces are connected in parallel, according to one embodiment.

Referring to FIG. 5, a continuous sedimentation-based microfluidic separation and culture system (hereinafter referred to as a system) 500 according to one embodiment includes a plurality of capture space portions 510, 520, and 530 connected in parallel. can do.

In this case, the length of each of the plurality of capture spaces 510, 520, 530 can be determined and adjusted based on the settling velocity and the dropping point of each of the plurality of target particles that the system 500 is intended to capture. For example, the first capture space 510 may set the sedimentation rate of the first target particle to be separated and captured among the particles included in the sample, and the sedimentation rate of each of the second and third target particles to pass through. By determining and adjusting the length in consideration of the above, only the first target particles may be selectively captured and separated, and the second capture space 520 may be configured to separate and capture the second target particles from among the particles included in the sample. The length is determined and adjusted in consideration of the sedimentation rate and the sedimentation rate of each of the first target particle and the third target particle to be passed, so that only the second target particle can be selectively captured and separated, and the third capture space portion ( 530 is a length in consideration of the sedimentation rate of the third target particles to be separated and captured among the particles included in the sample and the sedimentation speed of each of the first and second target particles to pass through. It is determined and adjusted, only the target particles 3 can be selectively captured and isolated from each other. More specifically, for example, the length of the first capture space 510 may be defined by a condition in which the settling time t _1S corresponding to the settling velocity of the first target particle is smaller than the passage time t _1T (t _1S <t _1T ) and the second target. Settling time t _2S according to the settling velocity of the particles is greater than the passing time t _2T (t _2S > t _2T ) and settling time t _3S is greater than the passing time t _3T according to the settling velocity of the third target particle (t _3S > t _3T ), and the length of the second capture space 520 is determined such that the settling time t _2S according to the settling velocity of the second target particle is smaller than the passing time t _2T (t _2S <t _2T ), the settling time t _1S according to the settling speed of the first target particle is greater than the passing time t _1T (t _1S > t _1T ) and the settling time t _3S according to the settling speed of the third target particle t _3T It is determined and adjusted to satisfy a larger condition (t _3S > t _3T ), the length of the third capture space 530 is the length of the third target particles Settling time t _3S according to the settling velocity is smaller than the passing time t _3T (t _3S <t _3T ), and settling time t _1S is larger than the passing time t _1T according to the settling velocity of the first target particle (t _1S > t _1T ) and the settling time t _3S according to the settling velocity of the third target particle may be determined and adjusted to satisfy a condition (t _3S > t _3T ) greater than the passage time t _3T .

In this case, each of the plurality of capture spaces 510, 520, and 530 may further include a capture medium (not shown) described above with reference to FIGS. 2A to 2B to continuously capture and culture the target particles. . In addition, each of the plurality of capture spaces 510, 520, and 530 may further include an optical sensor provided to observe the captured and cultured target particles.

As described above, a case in which each of the plurality of capture spaces 510, 520, and 530 captures and separates different target particles has been described. However, the present disclosure is not limited thereto, and the system 500 includes a plurality of capture spaces 510, 520. 530 may be implemented such that each captures the same target particle. This is to improve the throughput of capturing the target particles, in which case the length of each of the plurality of capture spaces 510, 520, 530 can all be determined and adjusted equally.

4 to 5, the length of each of the plurality of capture spaces 410, 420, 510, 520, and 530 is determined based on the settling velocity of each of the target particles 430 and 440 to be separated and captured. As described above with reference to FIGS. 1A-1C, it may be determined based on the drop points of each of the target particles 430 and 440 to be separated and captured. Detailed description thereof has been described above with reference to FIGS. 1A to 1C and will be omitted.

6A-6B illustrate a continuous sedimentation based microfluidic separation and culture system according to another embodiment. Specifically, FIG. 6A is a top view showing a continuous sedimentation-based microfluidic separation and culture system when the first inlet and the first outlet are opened, and FIG. 6B is a continuous settling-based microstructure when the second inlet and the second outlet are opened. Top view showing a fluid separation and culture system.

6A-6B, a continuous sedimentation-based microfluidic separation and culture system (hereinafter referred to as a system) 600 according to another embodiment is one such as the system 200 shown in FIGS. 2A-2B. Including the capture space portion 610, there is a difference in that the inlet and outlet are provided with two each.

More specifically, the system 600 is disposed in the first direction, the first inlet 620 through which the first sample flowing in the first direction is introduced, the first outlet 621 through which the first sample is discharged, and the first direction. The capture space 610 includes a second inlet 630 disposed in a second direction orthogonal to the second sample flowing in the second direction and a second outlet 631 through which the second sample is discharged. The first inlet 620, the second inlet 630, and the first outlet are disposed between the first inlet 620 and the first outlet 621, and between the second inlet 630 and the second outlet 631. At least one of the first sample is formed to have a height higher than the 621 and the second outlet 631, and flows from the first inlet 620 toward the first outlet 621 based on the hydraulic leap phenomenon At least one second target included in the second sample flowing from the first target particles and the second inlet 630 toward the second outlet 631. Each particle can be captured.

For example, when capturing at least one first target particle included in the first sample, the system 600 maintains the first inlet 620 and the first outlet 621 open as shown in FIG. 6A. The first inlet 630 and the second outlet 631 are maintained in the closed state to introduce the first sample into the capture space 610 so as to firstly utilize the hydraulic jumping phenomenon described above with reference to FIGS. 1A to 1C. The first target particles included in the sample may be captured and cultured in the first capture media 622 and 623. In this case, the first-first capture medium 622 of the first capture mediums 622 and 623 captures and incubates target particles having a high sedimentation rate and a closest drop point among the first target particles, and the first-second capture medium 622. The capture medium 623 may capture and culture target particles having a slow sedimentation rate and far drop points among the first target particles.

For another example, when capturing at least one second target particle included in the second sample, the system 600 maintains the second inlet 630 and the second outlet 631 open as shown in FIG. 6B. And the first inlet 620 and the first outlet 621 are kept closed so that the second sample is introduced into the capture space 610 so as to use the hydraulic jump phenomenon described above with reference to FIGS. 1A to 1C. The second target particles included in the second sample may be captured and cultured in the second capture media 632 and 633. At this time, the second-first capture medium 632 of the second capture mediums 632 and 633 captures and incubates the target particles having a high sedimentation rate and near the drop point among the second target particles, and second-2-2. The capture medium 633 may capture and culture target particles having a slow sedimentation rate and far drop points among the second target particles.

Accordingly, the length 611 of the capture space 610 in the first direction can be determined and adjusted based on the sedimentation velocity and the dropping point of the at least one first target particle included in the first sample, and the capture space The length 612 of the portion 610 in the second direction may be determined and adjusted based on the settling velocity and the dropping point of the at least one second target particle included in the second sample.

The system 600 having such a structure may have an advantage of miniaturization than a system including a plurality of capture spaces.

In the foregoing, a structure in which the system 600 includes capture media 622, 623, 632, and 633 has been described. In some embodiments, the capture media 622, 623, 632, and 633 may be omitted.

Although the embodiments have been described by the limited embodiments and the drawings as described above, various modifications and variations are possible to those skilled in the art from the above description. For example, the described techniques may be performed in a different order than the described method, and / or components of the described systems, structures, devices, circuits, etc. may be combined or combined in a different manner than the described method, or other components. Or, even if replaced or substituted by equivalents, an appropriate result can be achieved.

Therefore, other implementations, other embodiments, and equivalents to the claims are within the scope of the claims that follow.

Claims (18)

In the continuous sedimentation-based microfluidic separation and culture system using a hydraulic leap phenomenon,
An inlet through which a sample is introduced;
An outlet through which the sample is discharged; And
It is disposed between the inlet and the outlet and has a height higher than that of the inlet and the outlet, and captures at least one target particle included in the sample flowing from the inlet toward the outlet using a hydraulic jump phenomenon. Capture space part to say
Including,
The width of the inlet,
Determined to increase the rate of sample at the inlet to improve throughput of capturing the at least one target particle,
The width of the capture space portion,
Determined to reduce the velocity of the sample in the capture space to improve the capture rate for capturing the at least one target particle,
The hydraulic leap phenomenon,
When the sample flowing through the inlet enters the capture space portion includes the phenomenon that the at least one target particles rise and settle due to the rapid height change between the inlet and the capture space portion,
The capture space portion,
At least one capture medium for capturing and culturing the at least one target particle while being disposed at a dropping point of the at least one target particle in consideration of the settling velocity of the at least one target particle,
The volume and width of the capture space portion,
A continuous sedimentation based microfluidic separation and culture system determined based on microenvironmental requirements for culturing the at least one target particle in the at least one capture medium. The method of claim 1,
The capture space portion,
The sedimentation-based microfluidic separation and culture system of seizing the at least one target particles as the height of the capture space is sharply higher than the height of the inlet, the moving speed of the sample is lowered in the capture space. The method of claim 2,
The capture space portion,
Continuous sedimentation-based microparticles, which selectively capture the at least one target particle of the particles as the particles have different drop points and sedimentation rates based on the size or density of each of the particles included in the sample Fluid Separation and Culture System. delete The method of claim 1,
The continuous sedimentation-based microfluidic separation and culture system,
And further comprising an optical sensor for observing at least one target particle incubated in said at least one capture medium. The method of claim 3,
The length of the capture space portion,
And a sedimentation based microfluidic separation and culture system based on the sedimentation rate of the at least one target particle. delete In the sedimentation-based microfluidic separation and cultivation method using a hydraulic leap phenomenon,
Introducing a sample through an inlet;
At least one target particle included in the sample flowing from the inlet toward the outlet through a capture space formed between the inlet and the outlet through which the sample is discharged to have a height higher than that of the inlet and the outlet. Capturing using a hydraulic jump phenomenon; And
Discharging the sample captured by the at least one target particle through the outlet;
Including,
The introducing step is
Introducing the sample through an inlet whose width is determined to increase the speed of the sample at the inlet in order to improve the throughput of capturing the at least one target particle,
The capturing step,
Capturing the at least one target particle through the capture space portion whose width is determined to reduce the velocity of the sample in the capture space portion to improve the capture rate for capturing the at least one target particle,
The hydraulic leap phenomenon,
When the sample flowing through the inlet enters the capture space portion includes the phenomenon that the at least one target particles rise and settle due to the rapid height change between the inlet and the capture space portion,
The capturing step,
Capturing and incubating the at least one target particle through at least one capture medium disposed at a dropping point of the at least one target particle in consideration of the settling velocity of the at least one target particle
Including,
The volume and width of the capture space portion,
And a method for continuous sedimentation based microfluidic separation and culturing determined on the basis of microenvironmental requirements for culturing the at least one target particle in the at least one capture medium. The method of claim 8,
The capturing step,
Continuous sedimentation-based microfluidic separation and culture method of the step of seizing the at least one target particles as the height of the capture space is sharply higher than the height of the inlet, the moving speed of the sample is lowered in the capture space. . The method of claim 9,
The capturing step,
Continuous sedimentation, which selectively captures the at least one target particle of the particles as the particles have different drop points and settling rates based on the size or density of each of the particles included in the sample Based microfluidic separation and culture methods. delete In the continuous sedimentation-based microfluidic separation and culture system using a hydraulic leap phenomenon,
An inlet through which a sample is introduced;
An outlet through which the sample is discharged; And
A plurality of capture spaces disposed in series or in parallel between the inlet and the outlet, each of the plurality of capture spaces being formed to have a height higher than that of the inlet and the outlet and flowing toward the outlet from the inlet; Capture different target particles included in the ship using hydraulic leap
Including,
The width of the inlet,
Determined to increase the rate of sample at the inlet to improve throughput of capturing the different target particles,
The width of each of the plurality of capture spaces,
And to reduce the velocity of the sample in each of the plurality of capture spaces in order to improve the capture rate for capturing the different target particles,
The hydraulic leap phenomenon,
And when the sample flowing through the inlet enters each of the plurality of capture spaces, the different target particles rise and settle due to a sudden height change between the inlet and each of the plurality of capture spaces. ,
Each of the plurality of capture spaces,
A capture medium for capturing and culturing the different target particles while being disposed at falling points of the different target particles in consideration of the settling speeds of the different target particles;
The volume and width of each of the plurality of capture spaces,
Continuous sedimentation based microfluidic separation and culture system determined based on microenvironmental requirements for culturing the different target particles in the capture medium. The method of claim 12,
Each of the plurality of capture spaces,
A continuous sedimentation-based microfluid fluid that seizes and captures different target particles as the height of each of the plurality of capture spaces is sharply higher than the height of the inlet so that the moving speed of the sample decreases in each of the plurality of capture spaces. Separation and Culture System. The method of claim 13,
Each of the plurality of capture spaces,
Continuous sedimentation-based microfluid, which selectively captures the different target particles among the particles as the particles have different drop points and sedimentation rates based on the size or density of each of the particles included in the sample. Separation and Culture System. delete The method of claim 12,
The continuous sedimentation-based microfluidic separation and culture system,
And a plurality of optical sensors provided in correspondence with each of the plurality of capture spaces to observe the target particles incubated in the capture medium. The method of claim 14,
Each of the plurality of capture spaces,
Continuous sedimentation-based microfluidic separation and culture system is formed of different lengths based on the sedimentation rate of the different target particles. In the continuous sedimentation-based microfluidic separation and culture system using a hydraulic leap phenomenon,
A first inlet through which the first sample is introduced;
A first outlet through which the first sample is discharged;
A second inlet through which a second sample is introduced;
A second outlet through which the second sample is discharged; And
It is formed to have a height higher than the first inlet, the second inlet, the first outlet and the second outlet while being disposed between the first inlet and the first outlet and between the second inlet and the second outlet. And at least one first target particle included in the first sample flowing from the first inlet toward the first outlet and the second flow from the second inlet toward the second outlet based on a hydraulic jump phenomenon. Capture space portion for capturing each of the at least one second target particles contained in the second sample
Including,
The width of the first inlet,
A value that increases the speed of the first sample at the first inlet to improve throughput of capturing the at least one first target particle,
The width of the second inlet,
Determined to increase the speed of the second sample at the second inlet to improve throughput of capturing the at least one second target particle,
The width of the capture space portion,
And to decrease the speed of each of the first sample and the second sample in the capture space to improve the rate of capture of each of the at least one first target particle and the at least one second target particle. ,
The hydraulic leap phenomenon,
When the sample flowing through the first inlet enters the capture space, the at least one first target particle rises and sinks due to a sudden height change between the first inlet and the capture space; And when the sample flowing through the second inlet enters the capture space, the at least one second target particle rises and sinks due to a sudden height change between the second inlet and the capture space. ,
The capture space portion,
At least one first capture medium for capturing and culturing the at least one first target particle while being disposed at a dropping point of the at least one first target particle in consideration of the settling velocity of the at least one first target particle; At least one second capture medium for capturing and culturing the at least one second target particle while being disposed at a dropping point of the at least one second target particle in consideration of the settling velocity of the at least one second target particle Including,
The volume and width of the capture space portion,
Microenvironmental requirements for culturing the at least one first target particle in the at least one first capture medium and microenvironmental requirements for culturing the at least one second target particle in the at least one second capture medium. Continuous sedimentation based microfluidic separation and culture system determined on the basis of.

Images