KR102036723B1 - Intracytoplasmic sperm injection system and method of sperm injection - Google Patents

Abstract

According to an aspect of the present invention, the microfluidic chip includes at least one channel into which the oocytes and sperm are injected, and an inner space to which the oocytes and the sperm bind; And at least one manipulator movably coupled to the microfluidic chip and capable of steering in the interior space.

Description

Intracellular sperm injection system and sperm injection method using the same {INTRACYTOPLASMIC SPERM INJECTION SYSTEM AND METHOD OF SPERM INJECTION}

The present invention relates to an intracellular sperm injection (ICSI) system and a method for sperm injection using the same, and more particularly, a robotic intracellular sperm injection system and a sperm injection method incorporating a manipulator driven by a microfluidic chip and an automated system. It is about.

Intracellular Sperm Direct Injection (ICSI) is a type of in vitro fertilization (IVF) that involves artificially injecting a single sperm into an egg cell. It is widely used in fertilization of animals, including humans, for the purpose of overcoming male factor infertility or for the study of the underlying mechanism of fertilization. Since the inception of ICSI for the first time, this approach has led to very high levels of modification success.

ICSI is not only a very useful tool for overcoming certain types of male infertility, but also as a tool for fertilization or sperm-mediated gene transfer (abbreviated as SMGT) in experimental animals.

For two decades after 1992, ICSI was performed manually by a highly skilled technician adjusting the micromanipulator and the like while looking directly through the microscope's eyepiece. However, in the case of such manual operation, the yield varies depending on the skill and condition of the technician and has a relatively low modification rate of 50% to 80%.

In the past decade, efforts have been made to automate cell injection using automation and robotics approaches, but only a few steps are automated and expensive manipulators are still required to manually control the manipulator or minimize manual steps. And the problem of using stages.

U.S. Patent Publication No. 6,376,743 (April 23, 2002)

It is an object of the present invention to provide an intracellular sperm injection system and a sperm injection method capable of automating the ICSI process to achieve a high and uniform fertilization rate.

It is also an object of the present invention to provide an intracellular sperm injection system and a sperm injection method having low manufacturing cost by employing a manipulator capable of steering with a magnetic field.

In addition, it is an object of the present invention to provide an intracellular sperm injection system and a sperm injection method which can prevent contamination by using a sealed disposable microfluidic chip and can be used repeatedly by replacing only the disposable microfluidic chip.

However, these problems are exemplary, and the scope of the present invention is not limited thereby.

According to an aspect of the present invention, the microfluidic chip includes at least one channel into which the oocytes and sperm are injected, and an inner space to which the oocytes and the sperm bind; And at least one manipulator movably coupled to the microfluidic chip and capable of steering in the interior space.

The microfluidic chip may further include a support member for supporting the microfluidic chip, and the microfluidic chip may be attached to or detached from the support member.

The support member may include at least one channel connection portion corresponding to the at least one channel and at least one opening into which the at least one manipulator is inserted.

It may further include a front and rear movement means coupled to the at least one manipulator outside the support member to move the at least one manipulator.

The apparatus may further include a magnetic field generator configured to steer the manipulator.

The magnetic field generating device may include a plurality of coils, and the plurality of coils may be embedded in a supporting member supporting the microfluidic chip.

The manipulator may comprise a magnet.

The microfluidic chip may include an intake unit for supporting the egg cells by controlling air pressure.

According to another aspect of the present invention, the microfluidic chip comprising at least one channel into which the oocytes and sperm are injected and an inner space to which the oocytes and the sperm bind, and the microfluidic chip movably coupled to the microfluidic chip, A sperm injection method using an intracellular sperm injection system comprising at least one manipulator capable of steering, comprising: creating an environment of the interior space of the microfluidic chip; Injecting the egg cells through the at least one channel; Determining the location of the egg cell; Injecting the sperm through the at least one channel; Capturing the sperm; And injecting the sperm into the egg cells.

The manipulator includes a first manipulator and a second manipulator, performing the step of determining the position of the egg cell using the first manipulator, capturing the sperm using the second manipulator and Injecting the sperm may be performed.

The method may further include forming a hole in the egg cell between determining the location of the egg cell and injecting the sperm through the at least one channel.

The at least one channel includes a plurality of channels, and the solution and gas, the egg cell and the sperm may be injected through different channels, respectively, to create the environment.

The intracellular sperm injection system may further include a support member for supporting the microfluidic chip, and the microfluidic chip may be attached to or detached from the support member.

The intracellular sperm injection system may further include a front and rear movement means coupled to the manipulator to move the manipulator and a magnetic field generating device for steering the manipulator.

The magnetic field generating device may include a plurality of coils, and the plurality of coils may be embedded in a supporting member supporting the microfluidic chip.

Other aspects, features, and advantages other than those described above will become apparent from the following detailed description, claims, and drawings.

According to one embodiment of the present invention made as described above, it is possible to provide an intracellular sperm injection system and sperm injection method that can achieve a high and uniform fertilization rate by automating the ICSI process.

In addition, by employing a manipulator capable of steering with a magnetic field, it is possible to provide an intracellular sperm injection system and a sperm injection method having a low manufacturing cost.

In addition, it is possible to provide an intracellular sperm injection system and a sperm injection method which can prevent contamination by using a sealed disposable microfluidic chip and can be repeatedly used by replacing only the disposable microfluidic chip.

Of course, the scope of the present invention is not limited by these effects.

1 is an exploded perspective view schematically showing the components of the intracellular sperm injection system according to an embodiment of the present invention.
FIG. 2 is a perspective view showing an intracellular sperm injection system to which the components of FIG. 1 are combined. FIG.
3 is a schematic perspective view of the intracellular sperm injection system of FIG. 2 further comprising a magnetic field generating device and a microscope system.
4 is a flow chart sequentially illustrating the steps of the sperm injection method.
5A to 5H are conceptual views schematically illustrating a state of the microfluidic chip corresponding to each of the steps of FIG. 4.
6 is a perspective view schematically showing the components of another intracellular sperm injection system according to another embodiment of the present invention.
Figure 7 is a perspective view schematically showing the components of the intracellular sperm injection system according to another embodiment of the present invention.

DETAILED DESCRIPTION Various embodiments of the present disclosure are described below in connection with the accompanying drawings. Various embodiments of the present disclosure may have various changes and various embodiments, and specific embodiments are illustrated in the drawings and described in detail. However, this is not intended to limit the various embodiments of the present disclosure to specific embodiments, it should be understood to include all modifications and / or equivalents and substitutes included in the spirit and scope of the various embodiments of the present disclosure. In the description of the drawings, similar reference numerals are used for similar elements.

Expressions such as “comprises” or “can include” that can be used in various embodiments of the present disclosure indicate the existence of the corresponding function, operation or component disclosed, and additional one or more functions, operations or It does not restrict the components. In addition, in various embodiments of the present disclosure, the terms "comprise" or "have" are intended to indicate that there is a feature, number, step, action, component, part, or combination thereof described in the specification, It should be understood that it does not exclude in advance the possibility of the presence or addition of one or more other features or numbers, steps, operations, components, parts or combinations thereof.

In various embodiments of the present disclosure, an expression such as “or” includes any and all combinations of words listed together. For example, "A or B" may include A, may include B, or may include both A and B.

Expressions such as “first”, “second”, “first”, or “second”, etc. used in various embodiments of the present disclosure may modify various elements of the various embodiments, but do not limit the corresponding elements. Do not. For example, the above expressions do not limit the order and / or importance of the corresponding elements. The above expressions may be used to distinguish one component from another. For example, both a first user device and a second user device are user devices and represent different user devices. For example, without departing from the scope of the various embodiments of the present disclosure, the first component may be named a second component, and similarly, the second component may also be named the first component.

When a component is referred to as being "connected" or "connected" to another component, the component may be directly connected to or connected to the other component, It is to be understood that there may be new other components between the other components. On the other hand, when a component is referred to as being "directly connected" or "directly connected" to another component, it may be understood that there is no new other component between the component and the other component. Should be able.

The terms used in various embodiments of the present disclosure are merely used to describe specific embodiments, and are not intended to limit the various embodiments of the present disclosure. Singular expressions include plural expressions unless the context clearly indicates otherwise.

Unless defined otherwise, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which various embodiments of the present disclosure belong.

Terms such as those defined in the commonly used dictionaries should be construed as having meanings consistent with the meanings in the context of the related art, and are ideally or excessively formal, unless expressly defined in the various embodiments of the present disclosure. It is not interpreted in the sense.

1 is an exploded perspective view schematically showing the components of the intracellular sperm injection system according to an embodiment of the present invention, Figure 2 is a perspective view showing an intracellular sperm injection system combined with the components of FIG.

1 and 2, an intracytoplasmic sperm injection (ICSI) system 1 according to an embodiment may include at least an egg cell (OC, FIG. 5B) and a sperm (SM, FIG. 5F) injected therein. One space (P1, P2, P3, P4, P5, P6) is connected to the channels (P1, P2, P3, P4, P5, P6) and the inner space in which the oocyte (OC) and sperm (SM) bind ( And a microfluidic chip 110 including a 113 and at least one manipulator 130 movably coupled to the microfluidic chip 110 and capable of steering in the internal space.

The microfluidic chip 110 is a chip used to process or manipulate a small amount of fluid using a channel having a small size, and can easily control the injection and discharge of the fluid.

Although FIG. 1 illustrates that the microfluidic chip 110 includes six channels P1, P2, P3, P4, P5, and P6, the present invention is not limited thereto, and the channels P1, P2, P3, P4, The number of P5 and P6) is not particularly limited. The channel not only functions as an "inlet" into which solution, egg (OC), sperm (SM), gas and the like are injected, but also discharges solution, egg (OC), sperm (SM), gas, and lens. "Outlet". That is, a solution to be injected through at least one channel P1, P2, P3, P4, P5, or P6 may be injected or discharged.

According to an embodiment, the microfluidic chip 110 includes a plurality of channels P1, P2, P3, P4, P5, and P6, and each of the plurality of channels P1, P2, P3, P4, P5, and P6. Through the different materials can be injected or discharged. This configuration prevents contamination that may occur as different materials share channels P1, P2, P3, P4, P5, and P6. This will be described later.

The microfluidic chip 110 may further include a suction part SP for supporting the egg cell OC by controlling air pressure and a manipulator coupling part MP to which the at least one manipulator 130 is coupled.

Oocytes (OC) injected through the channels (P1, P2, P3, P4, P5, P6) is positioned by the suction unit (SP), and the channels (P1, By injecting sperm (SM) injected through P2, P3, P4, P5, P6 it is possible to combine the ovum cells (OC) and sperm (SM). As such, the space in which the ova cells OC and the sperm SM are coupled may be defined as the internal space 113 of the microfluidic chip 110.

According to one embodiment, the intracellular sperm injection system 1 may include a support member 120 for supporting the microfluidic chip 110. The support member 120 is a channel connection port (PC1, PC2, PC3, PC4, PC5, PC6) connected to the frame 121 and the channels (P1, P2, P3, P4, P5, P6) of the microfluidic chip 110. ) May be included. The frame 121 has openings H1, H2, H3, H4, H5, and H6 corresponding to the channel connection ports PC1, PC2, PC3, PC4, PC5, and PC6, and the channel connection ports PC1 and PC2. , PC3, PC4, PC5, and PC6 may be passages extending from the openings H1, H2, H3, H4, H5, and H6 of the frame 121. External egg cell sources (not shown) and sperm sources (not shown) are connected to the channel connection ports (PC1, PC2, PC3, PC4, PC5, PC6), and channel connection ports (PC1, PC2, PC3, PC4, PC5). Through the PC6, the egg cell OC and the sperm SM may be injected into the channels P1, P2, P3, P4, P5, and P6 of the microfluidic chip 100.

The support member 120 includes a suction part connecting port SPC connected to the suction part SP of the microfluidic chip 110, and the suction part connecting port SPC may be connected to an external pressure adjusting device (not shown). Can be. That is, the pressure of the suction part SP may be adjusted through the suction part connection port SPC using a pressure adjusting device (not shown). An opening H7 corresponding to the suction part connecting port SPC is formed in the frame 121, and the suction part connecting port SPC may be a passage extending from the opening H7 of the frame 121.

In addition, the support member 120 includes openings H8, H9 and H10 into which the manipulator 130 may be inserted, and the openings H8, H9 and H10 may be manipulator coupling portions of the microfluidic chip 110. It is disposed at a position corresponding to MP) and may have a number corresponding to the number of manipulators 130. The manipulator 130 may be inserted into the openings H8, H9, and H10 to be able to move back and forth.

Positioning of the egg cell (OC) and injection of sperm SM are performed using the manipulator 130, which is movably coupled to the microfluidic chip 110 to perform this operation. Steering may be possible at 113.

Although FIG. 1 illustrates that the manipulator 130 includes three, namely, a first manipulator 131, a second manipulator 132, and a third manipulator 131, 132, and 133, the present invention is not limited thereto. . That is, the manipulator may be one or two, or may be four or more. The first to third manipulators 131, 132, and 133 may each perform different functions in the sperm injection method.

The manipulator 130 is disposed to enable forward and backward movement and steering, and according to an embodiment, the forward and backward movement of the manipulator 130 may be performed by the forward and backward movement means 140 disposed outside the support member 120. have. For example, the front and rear movement means 140 may be a micro stage, and the first manipulator 131, the second manipulator 132, and the third manipulator 133 may be the first stage 141 and the second stage 142, respectively. And the third stage 143, respectively. Referring to FIG. 2, the first to third stages 141, 142, and 143 may be disposed in the control box 150, and the control box 150 may include the first to third steps 141, 142, and 143. Circuit portion (not shown) for controlling the driving of the) may be further disposed. The control box 150 may be connected to an external host computer (not shown) that includes a central processing unit.

According to an embodiment, steering of the manipulator 130 may be performed by control of the magnetic field. For steering by the magnetic field, the first manipulator 131, the second manipulator 132, and the third manipulators 131, 132, and 133 are respectively the first magnet M1, the second magnet M2, and the third manipulator. It may include a magnet (M3). This will be described later.

The microfluidic chip 110 may be mounted on or detached from the support member 120. For example, the microfluidic chip 110 may be disposable and may be replaced with another microfluidic chip 110 after use. The microfluidic chip 110 includes a lower plate 111 and an upper plate 112, and the channels P1, P2, P3, P4, P5, and P6 between the lower plate 111 and the upper plate 112 and the internal space 113. ) Is formed. The internal space 113 of the microfluidic chip 110 is substantially mounted from the external environment by the support member 120, the manipulator 130, the lower plate 111, and the upper plate 112 in a state where the microfluidic chip 110 is mounted on the support member 120. It can be kept closed.

Therefore, the microfluidic chip 110 is not easily contaminated from the external environment, and the microfluidic chip 110 contaminated by performing a sperm injection method may be replaced with a new microfluidic chip 110. Through this configuration, it is possible to prevent contamination of the inner space 113 to which the egg cell (OC) and sperm (SM) are coupled.

In addition, the intracellular sperm injection system 1 according to the embodiment has a simple configuration that forms an integrated platform in which the microfluidic chip 110, the support member 120, and the manipulator 130 are integrated, and the manufacturing cost is low. Commercialization can be easy.

3 is a schematic perspective view of the intracellular sperm injection system of FIG. 2 further comprising a magnetic field generating device and a microscope system.

Referring to FIG. 3, the intracellular sperm injection system 1 further includes a magnetic field generating device 160 and an imaging system 170, and the magnetic field generating device in the process of performing the intracellular sperm injection process. Steering manipulator 130 (FIG. 2) using 160 and microfluidic chip 110, egg cell (OC, FIG. 5B), sperm (SM, FIG. 5F) and manipulator 130 using imaging system 170. ) Can be taken and observed in real time.

For example, the magnetic field generating device 160 may include eight coils. Each coil generates a magnetic field, and the magnetic fields generated by the respective coils overlap to finally generate a magnetic field in the region of interest, that is, the internal space 113, and include magnets M1, M2, and M3 by adjusting them. The manipulator 130 may be steered about one axis. Although FIG. 3 illustrates that the magnetic field generating device 160 includes eight coils, the present invention is not limited thereto.

According to an embodiment, the imaging system 170 may be a microscope based system including a camera, and an image captured by the imaging system 170 may be transferred to a host computer and processed through a predetermined image processing algorithm. have. Based on the image processed in real time, the manipulator 130 may be adjusted to track and capture sperm SM and to be injected into the OC.

4 is a flow chart sequentially showing the steps of the sperm injection method, Figures 5a to 5h is a conceptual diagram schematically showing the state of the microfluidic chip corresponding to each of the steps of FIG.

Referring to FIG. 4, in the sperm injection method according to an embodiment, the environment of the internal space 113 of the microfluidic chip 110 may be generated (S111), channels P1, P2, P3, P4, P5, and P6. Injecting the oocyte (OC) through the step (S112), the step of determining the position of the egg cell (OC) using the first manipulator 131 (S113), channels (P1, P2, P3, P4, P5, Injecting sperm SM through P6) (S115), capturing sperm using the second manipulator 132 (S116), and sperm to the ovum cell OC using the second manipulator 132. Injecting (SM) may include the step (S117).

According to another embodiment, a step S114 of forming an opening in the egg cell OC using the third manipulator 133 between determining the position of the egg cell (S113) and injecting the sperm (S115) is performed. May be performed further.

Referring to FIG. 5A, before implanting the egg cells OC, forming the internal space 113 of the microfluidic chip 110 into an environment suitable for fertilization of the egg cells OC and sperm SM (S111). Can be done.

For example, a predetermined solution may be injected through one of the channels P1, P2, P3, P4, P5, and P6, and a predetermined gas may be injected through the other channel P6. For example, the solution may be a substance having a viscosity, and the gas may be dioxide gas (CO ₂ ) that can control the acidity of the solution. Dioxide (CO ₂ ) can be continuously injected while the steps of the sperm injection method are performed to keep the acidity of the solution constant. Although not shown, the intracellular sperm injection device may further include a heating unit (not shown), and the heating unit (not shown) may maintain the internal space 113 of the microfluidic chip 110 at a constant temperature, for example, about 37 degrees. Can be.

Referring to FIG. 4B, an operation of injecting oocytes OC through one of the channels P3 among the plurality of channels P1, P2, P3, P4, P5, and P6 may be performed (S112).

Referring to FIGS. 4C and 4D, the implanted egg cell OC is handled by using the first manipulator 131 to determine the position in the suction part SP having a relatively low pressure. (S113) may be performed.

The first manipulator 131 includes a first magnet M1 and steers and positions the egg cell OC by steering the first manipulator 131 including the first magnet M1 by a magnetic field. can do. For example, the first manipulator 131 may move back and forth by the first stage 141, and may be steered in a desired direction by manipulating the magnetic field of the magnetic field generating device 160 (FIG. 3).

Referring to FIG. 4E, a step (S114) of forming a hole in the egg cell OC fixed using the third manipulator 133 may be performed. The third manipulator 133 may include a third magnet M3, and may be steered by the third stage 143 by forward and backward movement and by a magnetic field.

Egg cell (OC) comprises an egg (OC _a), egg (OC _a) the surrounding zona pellucida (OC _b) and core (OC _c) with. In order to inject sperm (SM) without forming a hole in the egg (OC), an impact must be applied to penetrate the zona pellucida of the egg (OC _b ). These shocks and invasion can damage the egg cells (OC _b ).

According to one embodiment, the step of preforming holes may be performed while minimizing the impact on the oocytes (OC), and by performing these steps, damage to the oocytes (OC) may be minimized.

However, the present invention is not limited to this. That is, according to another embodiment, the step (S114) of forming a hole in the egg cell (OC) can be omitted.

Referring to FIG. 4F, a step S115 of injecting sperm SM through one channel P2 of the plurality of channels P1, P2, P3, P4, P5, and P6 may be performed.

Referring to FIG. 4G, the step S116 of capturing the sperm SM may be performed using the second manipulator 132. The second manipulator 132 may include a second magnet M2, and may be steered by the second stage 142 by forward and backward movement and by a magnetic field. The sperm SM may be tracked through analysis of an image captured by the imaging system 170, and may capture the sperm SM by moving the second manipulator 132 based on the tracked information. The second manipulator 132 includes an inner hole for trapping sperm, and a predetermined solution may be present inside the hole. In addition, the pressure of the inner hole of the second manipulator 132 is adjustable, it is possible to capture the sperm (SM) by providing a suction force through the pressure adjustment of the hole.

Referring to FIG. 4H, the captured sperm SM may be injected into the egg cell OC using the second manipulator 132.

The intracellular sperm injection may be performed through the steps of FIGS. 4A to 4H, and the fertilized egg in which the ova cells (OC) and the sperm (SM) are coupled to the channels P1, P2, P3, P4, P5, P6) may be discharged through at least one.

Referring to FIGS. 5A through 5H, a solution, a gas, and the like may be used by injecting different channels to inject a solution, a gas, an egg cell (OC), and a sperm (SM) into the internal space 113 of the microfluidic chip 110. Contamination that may occur during the injection of ovarian cells (OC), and sperm (SM) was minimized. However, the present invention is not so limited, and at least some of the injection material may share a channel.

6 is a perspective view schematically showing the components of another intracellular sperm injection system according to another embodiment of the present invention.

Hereinafter, the intracellular sperm injection system 2 according to an embodiment will be described with focus on the difference from the intracellular sperm injection system 1 of FIG. 2.

Referring to FIG. 6, the intracellular sperm injection system 2 according to an embodiment may include at least one channel P1, P2, P3, and P4 into which egg cells (OC, FIG. 5B) and sperm (SM, FIG. 5F) are injected. , Microfluidic chip 210 including P5 and P6 and internal space 213, and at least one manipulator 230 movably coupled to the microfluidic chip 210 and capable of steering in the internal space 213. It includes.

The intracellular sperm injection system 2 may include a support member 220 for supporting the microfluidic chip 210 and a magnetic field generating device 260 for steering the manipulator 230. According to an embodiment, the magnetic field generating device 260 including the plurality of coils may be embedded in the support member 220. That is, the magnetic field generating device 260 is not located outside the support member 220 as shown in FIG. 3, but is disposed in the support member 220 and is integrated with the microfluidic chip 210 and the manipulator 230. Can be formed. Although FIG. 6 shows that the coils are two pairs, the present invention is not limited thereto, and additional coils may be further disposed inside and / or outside the support member 220 to increase the degree of freedom in manipulator 230 steering.

Through such a configuration, it is possible to miniaturize the intracellular sperm injection system 2 and to increase the degree of freedom for the place where the intracellular sperm injection system 2 can be used.

According to an embodiment, the manipulator 230 may include a first manipulator 231, a second manipulator 232, and a third manipulator 233. As described above, the manipulator 230 may include a hole in the egg cell OC (FIG. 5B). When the step S114 of FIG. 4 is omitted, the third manipulator 233 may be omitted. The first manipulator 231, the second manipulator 232, and the third manipulator 233 may include a first magnet M1, a second magnet M2, and a third magnet M3, respectively.

Figure 7 is a perspective view schematically showing the components of the intracellular sperm injection system according to another embodiment of the present invention.

Referring to FIG. 7, the intracellular sperm injection system 3 according to an embodiment includes at least one channel P1 and P2 and an inner space into which an egg cell (OC, FIG. 5B) and a sperm (SM, FIG. 5F) are injected. A microfluidic chip 310 including a 313 and at least one manipulator 330 movably coupled to the microfluidic chip 310 and capable of steering in the internal space 313.

The intracellular sperm injection system 3 includes a support member 320 for supporting the microfluidic chip 310, which supports the channel connections PC1 and PC2 corresponding to the channels P1 and P2. It may include. The manipulator 330 may include a magnet M, and the intracellular sperm injection system 3 may include a magnetic field generating device 160 (FIG. 3) for steering the manipulator 3.

Since the microfluidic chip 310 of FIG. 7 includes fewer channels P1 and P2 than the microfluidic chip 110 of FIG. 2 and also includes only one manipulator 330, the microfluidic chip 310 may have a simpler configuration. In this case, at least some of the solution, gas, egg cells (OC, FIG. 5B) and sperm (SM, FIG. 5F) for the environmental composition may share the channels P1 and P2.

Manipulator 330 is to determine the location of the egg (OC, Figure 5b) (S113, Figure 4), forming a hole in the egg (OC, Figure 5b) (S114), capturing and injecting sperm (S116) It may be one manipulator 330 having a form capable of performing all of S117, or may be a generic name of another type of manipulator 330 that may be replaced according to a step.

Through the intracellular sperm injection system (1, 2, 3) and sperm injection method according to the embodiments described above, it is possible to automate the ICSI process to achieve a high and uniform fertilization rate. In addition, the manufacturing cost can be reduced by employing the manipulators 130, 230, and 330 that can steer with a magnetic field. In addition, by using the sealed disposable microfluidic chips 110, 210, and 310, contamination may be prevented and repeated use may be possible by replacing only the disposable microfluidic chip.

As described above, the present invention has been described with reference to the embodiments illustrated in the drawings, which are merely exemplary, and those skilled in the art will understand that various modifications and equivalent other embodiments are possible. . Therefore, the true technical protection scope of the present invention will be defined by the technical spirit of the appended claims.

1, 2, 3: intracellular sperm injection system
110, 210, 310: microfluidic chip
120, 220, 320: support member
130, 230, 330: manipulator
140: back and forth means of transportation
160, 260: magnetic field generating device
170: imaging system
P1, P2, P3, P4, P5, P6: Channel
SP: suction
MP: Manipulator Coupling
M1, M2, M3, M: Magnet

Claims (15)

A microfluidic chip comprising at least one channel into which the egg cells and sperm are injected and an inner space to which the egg cells and the sperm bind; And
At least one manipulator movably coupled to the microfluidic chip and capable of steering in the interior space;
Further comprising a support member for supporting the microfluidic chip,
The microfluidic chip may be attached to or detached from the support member.
The support member includes at least one channel connection portion corresponding to the at least one channel and at least one opening into which the at least one manipulator is inserted.
The at least one channel includes a plurality of channels,
Intracellular sperm injection system, the solution and gas for creating the environment of the inner space, the egg cell and the sperm are each injected through different channels. delete delete The method of claim 1,
Intracellular sperm injection system coupled to the at least one manipulator on the outside of the support member, further comprising a forward and backward movement means for moving the at least one manipulator. The method of claim 1,
And a magnetic field generating device for steering said manipulator. The method of claim 5,
The magnetic field generating device includes a plurality of coils,
And the plurality of coils are embedded in a support member for supporting the microfluidic chip. The method of claim 5,
And the manipulator comprises a magnet. The method of claim 1,
The microfluidic chip includes an inhalation portion that controls the air pressure to support the egg cells. A microfluidic chip including at least one channel into which the oocytes and sperm are injected, and an inner space in which the egg cells and the sperm are coupled, and at least one manipulator movably coupled to the microfluidic chip and capable of steering in the inner space. In the sperm injection method using an intracellular sperm injection system,
Creating an environment of the interior space of the microfluidic chip;
Injecting the egg cells through the at least one channel;
Determining the location of the egg cell;
Injecting the sperm through the at least one channel;
Capturing the sperm; And
Injecting the sperm into the egg cell,
The intracellular sperm injection system further includes a support member for supporting the microfluidic chip,
The microfluidic chip may be attached to or detached from the support member.
The support member includes at least one channel connection portion corresponding to the at least one channel and at least one opening into which the at least one manipulator is inserted.
The at least one channel includes a plurality of channels,
Solution and gas for the environment, the egg cell and the sperm are each injected through a different channel, sperm injection method.
The method of claim 9,
The manipulator includes a first manipulator and a second manipulator,
And determining the position of the egg cells using the first manipulator, capturing the sperm using the second manipulator, and injecting the sperm into the egg cells. The method of claim 9,
Between determining the location of the egg cells and injecting the sperm through the at least one channel,
Sperm injection method further comprising the step of forming a hole in the egg cell. delete delete The method of claim 9,
The intracellular sperm injection system further comprises a back and forth movement means coupled to the manipulator to move the manipulator and a magnetic field generating device for steering the manipulator. The method of claim 14,
The magnetic field generating device includes a plurality of coils,
And the plurality of coils are embedded in a support member for supporting the microfluidic chip.

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