JP2021069300A - Cell suction system, suction chip, and method for manufacturing suction chip - Google Patents

Abstract

PROBLEM TO BE SOLVED: To form a tip of a suction tip provided in a cell suction system 1 for sucking cells or at least a part of cells with a desired inner diameter. SOLUTION: The cell suction system 1 sucks a cell or at least a part of the cell as a suction target, and has a suction tip 3 provided with a take-in hole 306 capable of taking the suction target into an internal space. The operation of the suction device 4 capable of sucking the object to be sucked into the internal space of the suction tip with the suction tip attached, the transport unit 5 capable of moving the suction device in different three axial directions, and the operation of the transport unit. It is provided with a signal processing unit 6 that is controllable and can detect the position of the tip of the suction tip. The suction tip 3 has a fitting portion 305 that can be fitted with the suction portion of the suction device 4 while communicating with the intake hole 306, and is integrally molded with resin. [Selection diagram] Fig. 1

Description

The present invention relates to a cell aspiration system, a suction tip and a method for manufacturing the suction tip.

In research on life systems and the like, a work of identifying a characteristic cell from many cells in a cell culture vessel and sucking the cell or a part of the cell is frequently performed. Conventionally, a tubular suction tip is used for sucking cells or intracellular substances (see, for example, Patent Document 1).

The conventional suction tip is composed of a connector having a mounting port for a suction device and a glass tip tip, and the tip tip pulls the glass tube while heating the central part of the glass tube with a puller device. It is molded by tearing it while doing so. Therefore, it is difficult to form the tip of the suction tip into a desired inner diameter.

Japanese Unexamined Patent Publication No. 2019-33680

The present invention has been made in view of such conventional circumstances, and an object of the present invention is to form the tip of a suction tip into a desired inner diameter.

The cell suction system for solving the above-mentioned problems is a cell suction system that sucks cells or at least a part of the cells as the object to be sucked, and is provided with an intake hole capable of taking the object to be sucked into the internal space. A suction tip that can suck the object to be sucked into the internal space of the suction tip with the suction tip attached, and a transport unit that can move the suction device in different three axial directions. A signal processing unit capable of controlling the operation of the transport unit and detecting the position of the tip of the suction tip is provided, and the suction tip communicates with the suction hole and communicates with the suction unit of the suction device. It has a fitting portion that can be fitted and is integrally molded with a resin.

Further, the cell suction system includes a suction tip holder capable of holding the suction tip, and the suction tip is provided with a flange surface that comes into contact with the suction tip holder while being provided in the suction tip holder. Yes, the signal processing unit controls the operation of the transport unit, and by moving the suction device in one direction, the suction unit is pressed against the fitting portion, and the suction tip cage is pressed on the flange surface. The suction tip is attached to the suction device by obtaining a reaction force.

Further, in the cell suction system, the suction tip includes a base end portion provided with the fitting portion, a bottom portion connected to the base end portion, and a needle-shaped portion protruding from the bottom portion. The intake hole is provided so as to penetrate the bottom portion and the needle-shaped portion.

The suction tip for solving the above problems is a suction tip provided in the cell suction system according to any one of claims 1 to 3, and the suction tip is made of polypropylene, polycarbonate, or polystyrene. It is a feature.

The method for manufacturing a suction tip for solving the above problems is a method for manufacturing a suction tip, in which an outer mold for forming the outer shape of the suction tip and an inner mold for forming the inner shape of the suction tip are formed. The suction tip is integrally molded with a resin by using a mold and a rod-shaped core mold that is connected to the outer mold or the inner mold to form the intake hole.

According to the suction system, the suction tip, and the method for manufacturing the suction tip according to the present invention, the tip of the suction tip can be formed to have a desired inner diameter.

The figure which shows the structural example of the cell suction system which concerns on this invention. The figure which shows the structural example of the suction chip provided in the cell suction system which concerns on this invention. The figure which shows the structural example of the suction chip provided in the cell suction system which concerns on this invention. The figure which shows the structural example of the suction chip provided in the cell suction system which concerns on this invention. The figure which shows the structural example of the suction chip provided in the cell suction system which concerns on this invention. The figure which shows the structural example of the suction chip provided in the cell suction system which concerns on this invention. The figure which shows the application example of a light source device. The figure which shows the configuration example when there is a light receiving sensor in a confocal scanner.

The cell suction system according to the present invention sucks one cell or a part of the cell (intracellular substance). Hereinafter, as an example of the cell aspiration system according to the present invention, an embodiment in which one cell in a sample is aspirated will be described.

(Structure of Examples of Cell Aspiration System)
The cell aspiration system 1 shown in FIG. 1 includes a container (sample cage) 2 for storing the sample 201. Sample 201 contains cells to be aspirated and a culture medium for culturing the cells. The sample cage 2 is formed in a box shape having a bottom surface on which the sample 201 (cells) is placed and a side surface surrounding the bottom surface so that the sample 201 does not spill, and is installed so that the bottom surface is horizontal. ..

The cell suction system 1 includes a container (suction tip) 3 capable of taking in cells inside, a suction device 4 to which the suction tip 3 is mounted, and a transport unit 5 to which the suction device 4 is mounted. The cell suction system 1 includes a suction tip cage 300 (see FIG. 2) capable of individually holding a plurality of suction tips 3.

As shown in FIG. 2, the suction tip 3 has a base end portion 301 which is an end portion on the side to be mounted on the suction device 4, a tubular body portion 302 connected to the base end portion 301, and a body portion 302. A cone-shaped bottom portion 303 connected to the bottom portion 303 and a needle-shaped portion 304 protruding from the tip of the bottom portion 303 are provided.

The base end portion 301 includes a fitting hole (fitting portion) 305 into which the suction portion 401 of the suction device 4 is fitted, and the fitting hole 305 communicates with the internal space of the bottom portion 303 via the internal space of the body portion 302. To do. The suction tip 3 is included in the bottom portion 303 and the needle-shaped portion 304, and includes a thin tube hole (take-in hole) 306 provided so as to penetrate a part of the bottom portion 303 and the needle-shaped portion 304. Therefore, the fitting hole 305 communicates with the thin tube hole 306. The length of the needle-shaped portion 304 is about 1 mm. The length of the thin tube hole 306 in the longitudinal direction is 2 mm or less (for example, 1 mm to 2 mm). The suction chip provided in the cell suction system 1 is not limited to the one having the above configuration, and may have a configuration described in detail later as an application example.

The suction tip 3 is integrally molded with a resin (for example, a thermoplastic resin such as polypropylene, polycarbonate, polystyrene, etc.) using a mold. The mold used for molding the suction tip 3 includes an outer mold that forms the outer shape of the base end portion 301, the body portion 302, the bottom portion 303, and the needle-shaped portion 304, and the internal space of the fitting hole 305 and the body portion 302. It includes an inner mold that forms the inner space of the bottom portion 303 and an elongated rod-shaped core mold that is connected to the outer mold or the inner mold to form a capillary hole 306 (not shown). The suction tip 3 is preferably molded so that the thickness of each configuration of the base end portion 301, the body portion 302, the bottom portion 303, and the needle-shaped portion 304 is uniform. Further, the resin may be a thermosetting resin.

In a method of pouring resin into such a mold, that is, injection molding, a suction tip 3 having a thin tube hole 306 having a desired inner diameter can be molded by using core molds having different diameters. The capillary hole 306 has an inner diameter (about 1 μm to several hundred μm) capable of taking up cells from the opening on the tip surface of the needle-shaped portion 304. For example, the inner diameter of the capillary hole 306 may be 1 to several tens of μm, or may be 10 μm to several 100 μm. In the suction tip according to the present invention, cells or intracellular cells of various sizes are formed by forming a capillary hole 306 with an inner diameter smaller than the size of the object to be sucked, corresponding to the size of the object to be sucked. The substance can be properly taken into its internal space.

As shown in FIG. 1, the cell aspiration system 1 includes a signal processing unit 6 that is connected to the aspirator 4 so as to be able to transmit and receive, and the operation of the aspirator 4 is controlled by the signal processing unit 6. The aspirator 4 sucks cells into the suction tip 3 by using a change in pressure or the like, and the signal processing unit 6 sucks the cells in the sample holder 2 into the suction tip 3 and sucks the cells. The operation of ejecting cells from the suction chip 3 is controlled.

The transport unit 5 is connected to the signal processing unit 6 so as to be able to transmit and receive, and the cell suction system 1 controls the operation of the transport unit 5 by the signal processing unit 6. The transport unit 5 can move the suction tip 4 in different three axial directions by using a motor or the like (not shown), and the signal processing unit 6 moves the suction tip 3 housed in the suction tip holder 300 to the suction tip 4. The operation of attaching the cells to the aspirator 4 and the operation of moving the cells to a position where the cells can be aspirated by the aspirator 4 equipped with the suction tip 3 are controlled. Therefore, the signal processing unit 6 can move the suction tip 3 in different triaxial directions via the transport unit 5.

Further, the cell suction system 1 has a configuration (sample position detection mechanism) for detecting the position of the cell to be sucked in the sample 201 in order to suck the cell to be sucked, and the suction tip 3 It is provided with a configuration (chip tip position detection mechanism) for detecting the position of the tip (chip tip position). In the following, each of these configurations will be described in order.

First, in the cell aspiration system 1, the sample position detection mechanism detects the position of the cell by imaging the fluorescence emitted by the cell in the sample holder 2, and the light (excitation light) for the cell to emit fluorescence. ) Is provided as a light source device 7. The light source device 7 is installed below the sample holder 2 (the side opposite to the side where the suction tip 3, the suction device 4, and the transport unit 5 are installed), and three light sources having different wavelengths (internal light source 701). , 702, 703) and three mirrors 711, 712, 713 corresponding to the internal light sources 701 to 703.

The first mirror 711 is a dichroic mirror capable of reflecting the excitation light L1 emitted from the first internal light source 701 and transmitting light other than the excitation light L1. The second mirror 712 is a dichroic mirror capable of reflecting the excitation light L2 emitted from the second internal light source 702 and transmitting light other than the excitation light L2. The third mirror 713 is capable of reflecting the excitation light L3 emitted from the third internal light source 703. The mirrors 711 to 713 are installed so that the optical paths of the excitation lights L1 to L3 reflected by the reflection surfaces overlap. The mirrors 711 to 713 are installed so that the excitation lights L1 to L3 reflected by each reflecting surface are projected onto the sample 201.

The light source device 7 is connected to the signal processing unit 6 so as to be able to transmit and receive, and the excitation lights L1 to L3 output from the light source device 7 are controlled by the signal processing unit 6. Specifically, an internal light source capable of emitting excitation light having a wavelength suitable for the cells to fluoresce is selected, and the light emitted from the selected, for example, the internal light source 701 is used as the excitation light L1 through the mirror 711. Light is projected onto the sample 201 from the bottom surface side of the sample cage 2.

The cell suction system 1 includes a microscope 8 on the optical path from the sample holder 2 to the light source device 7. The microscope 8 includes an objective lens 801 and an imaging lens 802 in order from the one closest to the sample holder 2. The optical axes of the objective lens 801 and the imaging lens 802 coincide with each other, and the optical axes are arranged along the optical paths of the excitation lights L1 to L3. The objective lens 801 is provided so as to be movable in the optical axis direction thereof. Further, the microscope 8 is connected to the signal processing unit 6 so as to be able to transmit and receive, and the movement (position in the optical axis direction) of the objective lens 801 is independently controlled by the signal processing unit 6. That is, the focusing point by the objective lens 801 is adjusted to an appropriate position by the signal processing unit 6. By the adjustment by the signal processing unit 6, for example, the objective lens 801 collects the excitation lights L1 to L3 in the vicinity of the sample 201. The imaging lens 802 is installed at a predetermined position with respect to the confocal scanner 9 so as to collect fluorescence in the vicinity of the pinhole 901 described later.

The cell aspiration system 1 includes a Nipodisk-type confocal scanner 9 for obtaining a two-dimensional image of a cell to be aspirated. The confocal scanner 9 includes a disk-shaped pinhole disk 901 and a disk-shaped condensing disk 902 on the optical path from the light source device 7 to the microscope 8. A plurality of pinholes are spirally arranged on the pinhole disc 901, and the condensing disc 902 is provided with a plurality of microlenses in the same pattern as the pinholes on the pinhole disc 901. In the pinhole disk 901 and the condensing disk 902, the center (central axis) of each disk is coaxially connected, and each optical axis of the plurality of microlenses passes through each corresponding pinhole, and the objective lens 801 and the imaging are formed. It is parallel to the optical axis of the lens 802. The pinhole disk 901 and the condensing disk 902 are rotated by a motor (not shown) with the central axis as a rotation axis, and the excitation light L1 to L3 passes through two rotated disks (a plurality of microlenses and a plurality of pinholes). Is projected over a wide area (multiple locations) with respect to the projected object (cell).

Further, the confocal scanner 9 includes a dichroic mirror 903 on the optical path from the condensing disk 902 to the pinhole disk 901. The dichroic mirror 903 is capable of reflecting the fluorescence emitted by the cells through the excitation lights L1 to L3. Further, the confocal scanner 9 includes a camera 904 that captures the sample 201 on the fluorescent optical path reflected by the dichroic mirror 903. The camera 904 is installed so that the fluorescence reflected by the dichroic mirror 903 can be incident.

Further, the confocal scanner 9 includes a relay lens 905, a filter wheel 907 rotatably installed, and a relay lens 906 on the optical path from the dichroic mirror 903 to the camera 904. The filter wheel 907 includes four filters 917, 927, 937, 947 having different functions. The filters 917, 927, 937, 947 can be positioned on the optical path by the rotation of the filter wheel 907, and the light transmitted through the relay lens 905 is incident on the filter 917 or the filter 927. The filters 917, 927, 937, and 947 are all capable of transmitting light in different wavelength ranges. The filter 917 reflects or absorbs light other than the fluorescence due to the excitation light L1 reflected by the dichroic mirror 903, and can transmit the fluorescence. The filter 927 reflects or absorbs light other than the fluorescence due to the excitation light L2 reflected by the dichroic mirror 903, and can transmit the fluorescence. The filter 937 reflects or absorbs light other than the fluorescence due to the excitation light L3 reflected by the dichroic mirror 903, and can transmit the fluorescence. The filter 947 will be described later.

The relay lens 905 is arranged between the dichroic mirror 903 and the filter wheel 907 so that its optical axis is along the optical path and its focal point is aligned with the pinhole disk 901. The relay lens 906 is arranged between the filter wheel 907 and the camera 904 so that its optical axis coincides with the optical axis of the relay lens 905 and its focal point coincides with the imaging surface of the camera 904. The relay lens 906 collects the light transmitted through any of the filters (filters 917 to 947) provided on the filter wheel 907 to the camera 904.

The confocal scanner 9 having the above-described configuration is connected to the signal processing unit 6 so as to be able to transmit and receive. Therefore, in the confocal scanner 9, the signal processing unit 6 independently controls the rotation operation of the pinhole disk 901 and the condensing disk 902, the rotation operation of the filter wheel 907, the imaging operation of the camera 904, and the like. By rotating the filter wheel 907, filters (filters 917 to 947) located on the optical path of fluorescence reflected by the dichroic mirror 903 are selected. The image of the object (cell) to be sucked from the vertical direction captured by the camera 904 is recorded in the recording unit built in the camera 904. Then, the image is transferred to the signal processing unit 6 for analysis. The image is recorded not only in the recording unit inside the camera 904 but also in an external recording unit.

As described above, the cell suction system 1 is provided with the sample position detection mechanism, so that the fluorescence can be captured by the camera 904 to acquire an image of the cell, and the position of the cell can be detected from the analysis result of the image. .. In the present embodiment, the sample position detection mechanism includes a signal processing unit 6, a light source device 7, a microscope 8, and a confocal scanner 9.

Next, in the cell suction system 1, the chip tip position detection mechanism projects a predetermined light (detection light) to the tip of the suction tip 3 and reflects the detection light (detection reflected light) at the tip of the suction tip 3. The tip position of the chip is detected from the strength of. The tip tip position detecting mechanism includes a tip tip detecting unit 10 that acquires the position information of the tip of the suction tip 3. The chip tip detection unit 10 is provided on the optical path from the light source device 7 to the confocal scanner 9.

The chip tip detection unit 10 is a light source (chip tip detection light source) 101 that emits detection light.
A collimating lens 102 that makes the detected light parallel, a dichroic mirror 103 that has a function of reflecting the detected light and the detected reflected light and transmitting the excitation light L1 to L3, and the detected reflected light that is transmitted through the detected light. A half mirror 104 capable of reflecting light is provided. The dichroic mirror 103 is installed on an extension of the optical axis of the objective lens 801 and the imaging lens 802.

The chip tip detection light source 101 transmits the detection light emitted from the chip tip detection light source 101 through the half mirror 104 after being incident on the collimating lens 102 to become parallel light, and is reflected by the dichroic mirror 103 before being cofocal. The condensing disk 902 (microlens) in the scanner 9, the dichroic mirror 903 having the function of transmitting the detected light and the detected reflected light in addition to the above functions, and the pinhole disk 901 (pinhole) are passed through in order, and the objective lens 801 and the objective lens 801 are passed. It is installed along the optical axis of the imaging lens 802.

Further, the chip tip detection unit 10 includes a light receiving sensor 105 that measures the intensity of the detected reflected light reflected by the half mirror 104. The light receiving sensor 105 is installed so that the detected reflected light reflected by the half mirror 104 can be incident.

The chip tip detection unit 10 having the above-described configuration is connected to the signal processing unit 6 so as to be able to transmit and receive. Therefore, in the chip tip detection unit 10, the signal processing unit 6 independently controls the measurement operation of the light receiving sensor 105 and the like. The signal processing unit 6 detects the chip tip position (z position of the chip tip) in the vertical direction from the measurement result by the light receiving sensor 105.

Further, the chip tip detection unit 10 is provided with a zoom lens or an aperture mechanism on the optical path from the chip tip detection light source 101 to the collimating lens 102, and can adjust the luminous flux diameter of the detection light incident on the collimating lens 102, for example. (Not shown). By increasing the luminous flux diameter of the detection light incident on the collimating lens 102, the intensity of the light projected per unit area of the sample 201 can be reduced. On the other hand, by reducing the luminous flux diameter of the detection light incident on the collimating lens 102, the intensity of the detection light per unit area projected on the tip of the suction chip 3 increases, and the intensity of the detection reflected light increases. Therefore, the intensity ratio of the detected reflected light to noise (stray light from the outside, etc.) becomes large.

In the present embodiment, the dichroic mirror 903 has a function of reflecting the detected reflected light in addition to the above-mentioned function. That is, the dichroic mirror 903 can branch the detected reflected light into the reflected light and the transmitted light. The filter 927 is capable of reflecting or absorbing light other than the detected reflected light reflected by the dichroic mirror 903 and transmitting the detected reflected light. In the cell suction system 1, by positioning the filter 927 on the optical path by rotating the filter wheel 907, the tip of the suction tip 3 can be imaged from the vertical direction using the camera 904. The captured image is analyzed by the signal processing unit 6, and the tip position (x, y position of the tip tip) of the suction tip 3 in the horizontal direction is detected from the result.

As described above, the cell suction system 1 can detect the x, y, and z positions of the chip tip position by providing the chip tip position detecting mechanism. In the present embodiment, the z position of the chip tip may be detected by taking an image with a camera without detecting the z position of the chip tip by the chip tip position detecting mechanism. In this case, the z position of the tip of the suction tip 3 is detected based on the change in the contrast of the image showing the tip of the suction tip 3 and the image around it. The chip tip position detection mechanism includes a suction chip 3, a suction device 4, a transport unit 5, a signal processing unit 6, a microscope 8, a confocal scanner 9, and a chip tip detection unit 10. ..

(Action and operation of examples of cell aspiration system)
The operation and operation of the cell suction system 1 in the above configuration will be described below (see FIG. 1).

The cell aspiration system 1 detects the tip tip position and the cell position. Based on the position information of the tip of the suction tip 3 and the position information of the cells, the transport unit 5 moves the suction tip 3 together with the suction device 4 and sucks, for example, one cell in the sample holder 2 into the suction tip 3. The specific operation will be described below.

First, the cell suction system 1 attaches the suction tip 3 to the suction device 4. Specifically, the signal processing unit 6 controls the operation of the transport unit 5 to move the suction device 4 onto the hole of the suction tip holder 300, and then brings the suction tip 3 closer to the suction tip holder 300. Lower it. When the suction device 4 is lowered, the suction unit 401 is pressed against the fitting hole 304, and the reaction force from the sample cage 300 is on the surface (flange surface) 307 where the base end portion 301 and the suction tip holder 300 are in contact with each other. In response to this, the suction tip 3 is fitted into the suction portion 401.

After that, the signal processing unit 6 controls the operation of the transport unit 5 to move the suction chip 3 to the vicinity of the extension line of the optical axis of the objective lens 801 and the imaging lens 802.

Next, the cell aspiration system 1 detects the tip tip position. Specifically, the signal processing unit 6 controls the operation of the chip tip position detection mechanism to emit detection light from the chip tip detection light source 101 and receive the reflected light (detection reflected light) at the tip of the suction chip 3. By receiving it with the sensor 105, the z position at the tip of the chip is first detected. The detection light emitted from the chip tip detection light source 101 is made parallel light by the collimated lens 102 and is incident on the half mirror 104. Since the half mirror 104 has a function of transmitting the detected light and reflecting the detected reflected light, the detected light passes through the half mirror 104 as parallel light and is incident on the dichroic mirror 103.

Since the dichroic mirror 103 has a function of reflecting the detection light, the detection light of the parallel light transmitted through the half mirror 104 is reflected by the dichroic mirror 103, and the condensing disk 902 (microlens) in the confocal scanner 9 is used. , Dichroic mirror 903, and pinhole disk 901 (pinhole) are transmitted in this order.

Since the condensing disk 902 and the pinhole disk 901 are constantly rotated by the signal processing unit 6, the detection light condensed in the vicinity of the suction chip 3 by the objective lens 801 becomes a plurality of luminous fluxes on the suction chip 3. It is flooded. Therefore, even if the tip of the suction tip 3 is located at a position shifted in the horizontal direction with respect to the optical path of the detection light overlapping the optical axes of the imaging lens 802 and the objective lens 801, at least one of these plurality of luminous fluxes is 1. One light flux is projected onto the tip of the suction chip 3 to obtain the detected reflected light. That is, since each luminous flux of the detection light focused in the vicinity of the suction chip 3 is focused at a plurality of locations near the tip of the suction chip 3, the position of the tip of the suction chip 3 can be detected as compared with the prior art. Range is expanded.

The detection light (detection reflected light) reflected by the tip of the suction tip 3 passes through the objective lens 801 and the imaging lens 802 and the pinhole disk 901 (pinhole), and is incident on the dichroic mirror 903. Since the dichroic mirror 903 has a function of branching the detected reflected light into the reflected light and the transmitted light, the detected reflected light is branched into the reflected light and the transmitted light by the dicroic mirror 903.

The detected reflected light transmitted through the dichroic mirror 903 passes through the condensing disk 902, is reflected by the dichroic mirror 103 and the half mirror 104, and is incident on the light receiving sensor 105. The light receiving sensor 105 measures the intensity of the incident detected reflected light. The signal processing unit 6 measures the intensity of the detected reflected light while moving the suction chip 3 in the vertical direction by the transport unit 5, and determines the position of the suction chip 3 when the measured intensity of the detected light is maximum. The z position at the tip of the chip.

On the other hand, the detected reflected light reflected by the dichroic mirror 903 is incident on the relay lens 905. The detected reflected light that has become parallel light by the relay lens 905 is incident on the filter wheel 907. At this time, the filter wheel 907 is in a state of being rotated so that the filter 947 is located on the optical path by the signal processing unit 6, and the detected reflected light is incident on the filter 947. Then, only the light in a predetermined wavelength range, that is, the detected reflected light is incident on the relay lens 906 by the filter 947. The detected reflected light collected by the relay lens 906 is incident on the camera 904. The camera 904 images the tip of the suction chip 3 with the detected reflected light received, and the image is recorded in the recording unit of the camera 904 and sent to the signal processing unit 6. The signal processing unit 6 analyzes the image and detects the x and y positions of the tip of the chip.

The tip position is detected by the above procedure. Using the above-mentioned camera, the z position of the tip of the suction tip 3 may be detected based on the change in the contrast of the image showing the image of the tip of the suction tip 3 and its surroundings in the same manner as the above-mentioned procedure.

The cell aspiration system 1 then detects the location of the cells. Specifically, the signal processing unit 6 operates and controls the sample position detection mechanism to select the light output from the light source device 7, that is, any of the internal light sources 701 to 703. The light selected here is light having a wavelength suitable for the cells to fluoresce, and for example, the internal light source 701 is selected. The excitation light L1 emitted from the selected internal light source 701 is reflected by the mirror 711 and output from the light source device 7.

The excitation light L1 output from the light source device 7 is incident on the dichroic mirror 103. Since the dichroic mirror 103 has a function of transmitting the excitation light L1, the excitation light L1 transmits the dichroic mirror 103 and transmits the condensing disk 902 (microlens), the dichroic mirror 903, and the pinhole in the confocal scanner 9. The disc 901 (pinhole) is passed through in order.

The light collecting disk 902 and the pinhole disk 901 are constantly rotated by the signal processing unit 6. The excitation light L1 that has passed through the microlens and the pinhole is incident on the imaging lens 802 and the objective lens 801 in this order and is projected onto the cells. The excitation light L1 focused in the vicinity of the sample 201 by the objective lens 801 becomes a plurality of luminous fluxes and is projected onto the sample 201 by the rotation of the focusing disk 902 and the pinhole disk 901.

The fluorescence emitted by the cells by the excitation light L1 passes through the objective lens 801 and the imaging lens 802 and the pinhole disk 901 (pinhole), and is incident on the dichroic mirror 903. Since the dichroic mirror 903 has a function of reflecting fluorescence, the fluorescence is reflected by the dichroic mirror 903.

The fluorescence from the excitation light L1 reflected by the dichroic mirror 903 is incident on the relay lens 905. The fluorescence that becomes parallel light by the relay lens 905 is incident on the filter wheel 907. At this time, the filter wheel 907 is in a state of being rotated so that the filter 917 is located on the optical path by the signal processing unit 6, and the fluorescence is incident on the filter 917. Then, only light in a predetermined wavelength range, that is, fluorescence, is incident on the relay lens 906 by the filter 917. The fluorescence collected by the relay lens 906 is incident on the camera 904. The camera 904 images the cells by the received fluorescence, and the image is recorded in the recording unit of the camera 904 and sent to the signal processing unit 6. The signal processing unit 6 analyzes the image and detects the position of the cell. Since the luminous flux of the excitation light L1 focused in the vicinity of the sample 201 is focused at a plurality of locations in the sample 201, it is possible to obtain an image having sufficient brightness required for image analysis.

By the above procedure, the position of the cell is detected. Even when the signal processing unit 6 selects the internal light source 702 or the internal light source 703, the position of the cell is detected by the same method.

In the cell suction system 1, the signal processing unit 6 identifies the suction target (cell) based on the cell position information, controls the transport unit 5 based on the tip position information of the suction tip 3, and controls the tip of the suction tip 3. Is moved onto the position of the object to be sucked. The aspirator 4 sucks the cells in the sample holder 2 into the suction tip 3 by the signal processing unit 6.

According to the cell aspiration system 1 according to the present embodiment, the suction tip 3 is formed by forming a capillary hole 306 with an inner diameter smaller than the size of the object to be aspirated, corresponding to the size of the object to be aspirated. , It is possible to take up cells of various sizes or intracellular substances into its internal space. Further, the suction tip 3 is integrally molded with resin using a mold, and a suction tip 3 having a thin tube hole 306 having a desired inner diameter can be molded by using core molds having different diameters. Can be done.

(Application example of the embodiment of the present invention)
A modified example of the above configuration is shown below.

The cell suction system according to the present invention is not limited to the one provided with the suction tip 3 like the cell suction system 1 described in the above-described embodiment. For example, the following 3a to 3d are used instead of the suction tip 3. It may be provided with.

The suction tip 3a shown in FIG. 3 has a configuration similar to that of the suction tip 3 except that the bottom is a flat plate instead of a cone, as compared with the suction tip 3 shown in FIG. The suction tip 3a includes a base end portion 301a, a body portion 302a, a needle-shaped portion 304a, a fitting hole 305a, and a thin tube hole 306a. The suction tip 3a is included in the bottom portion 303a and the needle-shaped portion 304a, and includes a thin tube hole (take-in hole) 306a provided so as to penetrate a part of the bottom portion 303a and the needle-shaped portion 304a. Therefore, the fitting hole 305a communicates with the thin tube hole 306a.

The suction tip 3b shown in FIG. 4 has the same configuration as the suction tip 3 except that it does not have a needle-shaped portion as compared with the suction tip 3 shown in FIG. The suction tip 3b includes a base end portion 301b, a body portion 302b, a bottom portion 303b, a fitting hole 305b, and a thin tube hole 306b. Since the suction tip 3b does not have a needle-shaped portion, the thin tube hole 306b includes a thin tube hole (take-in hole) 306b that is contained only in the bottom portion 303b and is provided so as to penetrate the bottom portion 303b. Therefore, the fitting hole 305b communicates with the thin tube hole 306b. The tip of the bottom 303b is thicker than the bottom 303 shown in FIG. 2 so that the length of the capillary hole 306b is substantially the same as the length of the capillary hole 306 shown in FIG. 2 (length suitable for taking up cells). It is molded.

Compared with the suction tip 3 shown in FIG. 2, the suction tip 3c shown in FIG. 5 has the same configuration as the suction tip 3 except that the base end portion is directly connected to the bottom portion without passing through the body portion. Have. The suction tip 3c includes a base end portion 301c, a bottom portion 303c, a needle-shaped portion 304c, a fitting hole 305c, a thin tube hole 306c, and a thin tube hole 306c. The suction tip 3c is included in the bottom portion 303c and the needle-shaped portion 304c, and includes a thin tube hole (take-in hole) 306c provided so as to penetrate a part of the bottom portion 303c and the needle-shaped portion 304c. Therefore, the fitting hole 305c communicates with the thin tube hole 306c. Further, since the suction tip 3c does not have a body portion, the base end portion 301c and the tapered portion 303c are directly connected.

Compared to the suction tip 3 shown in FIG. 2, the suction tip 3d shown in FIG. 6 does not have a needle-shaped portion, and the base end portion is directly connected to the bottom portion without passing through the body portion. It has the same configuration as the suction tip 3 except that. The suction tip 3d includes a base end portion 301d, a bottom portion 302d, a fitting hole 305d, and a thin tube hole 306d. Since the suction tip 3d does not have a needle-shaped portion, it includes a thin tube hole (take-in hole) 306d that is included in the bottom portion 303d and is provided so as to penetrate a part of the bottom portion 303d. Therefore, the fitting hole 305d communicates with the thin tube hole 306d. Further, since the suction tip 3d does not have a body portion, the base end portion 301d and the tapered portion 303d are directly connected. The tip of the bottom 303d is thicker than the bottom 303 shown in FIG. 2 so that the length of the capillary hole 306d is substantially the same as the length of the capillary hole 306 shown in FIG. 2 (length suitable for taking up cells). It is molded.

The cell aspiration system according to the present invention is not limited to the one provided with the light source device 7 like the cell aspiration system 1 described in the above-described embodiment.

In the cell suction system according to the present invention, the light source device may be any as long as it can output appropriate light to be projected onto the sample, and the number of internal light sources provided in the light source device is not particularly limited. At this time, the light source device may include one or more mirrors having appropriate properties depending on the arrangement of the internal light sources and the like.

For example, the cell suction system according to the present invention may have the following configurations such as 7e instead of the light source device 7. As shown in FIG. 7, the light source device 7e may be provided with only two dichroic mirrors (711e, 712e) for three internal light sources (701e, 702e, 703e). The internal light sources 701e and 702e and the dichroic mirrors 711e and 712e are installed in the same manner as in the present embodiment, and the internal light source 703e is on a straight line connecting substantially the centers of the reflection surfaces of the dichroic mirrors 711e and 712a and is an objective lens. It is installed on an extension of the optical axis of the 801 and the imaging lens 802.

Further, in the cell suction system according to the present invention, the light source device may include, for example, a tunable light source in which the wavelength of the emitted light is variable, in place of or in addition to the internal light source. When only a tunable light source is provided, the light source device does not have to be provided with a mirror.

The cell suction system according to the present invention is not limited to the cell suction system 1 described in the above-described embodiment, in which one rotatable filter wheel 907 is provided with a filter 917 and a filter 927. For example, instead of the filter wheel 907, the following configuration may be provided.

In the cell suction system according to the present invention, the structure and mechanism of the filter provided in front of the camera are not particularly limited as long as the wavelength of the light incident on the camera can be selectively limited. For example, in the cell suction system according to the present invention, separate filter wheels may be provided with filters having different functions, and independently slidable sliders or the like may be provided with filters having different functions. Each may be provided. The type of filter provided in the filter wheel corresponds to the type of light having a wavelength output from the light source device.

The cell suction system according to the present invention is not limited to the one in which the chip tip detection light source 101 is provided in the chip tip detection unit 10 as in the cell suction system 1 described in the above-described embodiment, and the suction chip It may be provided outside the chip tip detection unit as long as it is provided at a position where light can be projected on the tip. For example, the chip tip detection light source may be provided in the light source device, that is, the light source device may output not only the excitation light but also the detection light. In such a light source device, the chip tip detection light source is installed so that the detection light emitted from the tip detection light source is projected onto the tip of the suction chip after passing through the microlens and the pinhole.

The cell suction system according to the present invention is not limited to the one in which the light receiving sensor 105 is provided in the chip tip detection unit 10 as in the cell suction system 1 described in the above-described embodiment, and the light receiving sensor is a suction chip. If the intensity of the light reflected by the tip of the chip can be measured, it may be provided outside the chip tip detection unit. For example, as shown in FIG. 8, the light receiving sensor 909f may be installed in the confocal scanner 9f. At this time, the confocal scanner 9f includes a dichroic mirror 908f on the optical path from the dichroic mirror 903f to the relay lens 905f. The dichroic mirror 908f can branch _{the light L R1} reflected by the dichroic mirror 903f into the reflected light L _R2 and the transmitted light L _R3. The light receiving sensor 909f is installed so that the _{light L R2 reflected by the dichroic mirror 908f can be incident.}

In the cell aspiration system according to the present invention, regardless of any of the above-described modifications, the suction tip corresponds to the size of the object to be aspirated, as in the embodiment. By forming a capillary hole with an inner diameter smaller than the size of an object, cells of various sizes or intracellular substances can be taken into the internal space thereof. Further, the cell aspiration system according to the present invention is capable of aspirating intracellular substances even if it is not one cell shown in the examples.

1 Cell suction system 2 Sample cage 201 Sample 3 Suction tip 300 Suction tip cage 301 Base end 302 Body 303 Bottom 304 Needle 305 Fitting hole (fitting part)
306 Thin tube hole (take-in hole)
307 Flange surface 4 Aspirator 401 Aspirator 5 Conveyor 6 Signal processing unit 7 Light source device 701, 702, 703 Internal light source 711, 712, 713 Mirror 8 Microscope 801 Objective lens 802 Imaging lens 9 Confocal scanner 901 Pinhole disk 902 Condensing disk 903 Dycroic mirror 904 Camera 905 Relay lens 906 Relay lens 907 Filter wheel 917, 927, 937, 947 Filter 10 Chip tip detection unit 101 Chip tip detection light source 102 Collimating lens 103 Dycroic mirror 104 Half mirror 105 Light receiving sensor

Claims (5)

A cell aspiration system that aspirates cells or at least a portion of cells as an object to be aspirated.
A suction tip with a suction hole that allows the object to be sucked to be taken into the internal space,
A suction device to which the suction tip is attached and capable of sucking an object to be sucked into the internal space of the suction tip,
A transport unit that can move the aspirator in different three axial directions,
It is provided with a signal processing unit capable of controlling the operation of the transport unit and detecting the position of the tip of the suction tip.
The cell suction system is characterized in that the suction tip has a fitting portion that can be fitted with the suction portion of the suction device while communicating with the intake hole and is integrally molded with a resin. A suction tip holder capable of holding the suction tip is provided.
The suction tip is provided with a flange surface that comes into contact with the suction tip holder while being provided on the suction tip holder.
The signal processing unit controls the operation of the transport unit, pushes the suction unit against the fitting portion by moving the suction device in one direction, and counteracts from the suction tip holder on the flange surface. The cell suction system according to claim 1, wherein the suction tip is attached to the suction device by obtaining a force. The suction tip includes a base end portion provided with the fitting portion, a bottom portion connected to the base end portion, and a needle-shaped portion protruding from the bottom portion.
The cell suction system according to claim 1 or 2, wherein the intake hole is provided so as to penetrate the bottom portion and the needle-shaped portion. A suction chip provided in the cell suction system according to any one of claims 1 to 3.
The suction tip is a suction tip made of polypropylene or polycarbonate or polystyrene. The chip manufacturing method for manufacturing the suction tip according to claim 4.
An outer mold that forms the outer shape of the suction tip, an inner mold that forms the inner shape of the suction tip, and a rod-shaped core mold that is connected to the outer mold or the inner mold to form the intake hole. A chip manufacturing method, characterized in that the suction tip is integrally molded with a resin.

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