JP2018153142A - Nanopipette - Google Patents

Abstract

PROBLEM TO BE SOLVED: To easily and surely collect a desired object without imposing a load on the object. SOLUTION: A pipette body 2 having a first suction passage 10 capable of sucking an object C is formed inside, and a Co—Ni group alloy containing Co, Ni, Cr and Mo is formed, and the base end is formed on the pipette body. A pipette portion 3 to which the portion 21 is fixed is provided, and the pipette portion is formed inside with a second suction passage 22 capable of sucking an object communicating with the first suction passage and opening to the tip portion 23. Provided is a nanopipette 1 provided with a suction portion 20 and a protrusion 25 formed so as to protrude from the tip portion of the suction portion and elastically deformable in the radial direction orthogonal to the central axis. [Selection diagram] Fig. 6

Description

  The present invention relates to a nanopipette.

  In general, in the fields of biology (molecular biology, biochemistry, cell biology, etc.), drug discovery, drug development, etc., a biological sample obtained from a human body, an animal, a plant, etc., for example, a living cell can be cultured. Has been done. That is, separating cells from multicellular organisms and culturing the cells in vitro is performed. In performing such culture, for example, various culture methods are selected according to the type of cell, the purpose of culture, and the like. For example, the cells may be grown while the cells are attached to the culture vessel. . Such a cell is known as a so-called cultured cell, more specifically, an adherent culture cell, but a colony-like cell mass in which a plurality of cells are aggregated by the cells being connected to each other as they proliferate. It is easy to become.

Therefore, when cell passage is performed or when it is desired to separate only desired cells from the cell mass, it is required to break the cell mass apart and separate it into individual cells. In this case, as a conventional method, for example, a method of crushing a cell mass using the tip of a nanopipette and collecting a desired cell with the nanopipette is known.
For example, Patent Document 1 below discloses a nanopipette that can break a cell mass in a nanopipette by collecting a relatively small cell mass in a nanopipette and then repeatedly performing a suction operation and a discharge operation. Yes.

Japanese Patent No. 6004149

  However, in the conventional nanopipette, the cell mass is crushed by repeatedly performing the suction operation and the discharge operation, so that a load (damage) is likely to act on the cells. Therefore, even if the cell mass is crushed and separated into individual cells, loads are accumulated in these cells, and for example, it becomes difficult to collect the cells alive. Furthermore, even if the cell mass can be crushed, it is difficult to select and separate only desired cells, and it is difficult to reliably collect only desired cells with high reproducibility.

  The present invention has been made in view of such circumstances, and an object thereof is to provide a nanopipette capable of easily and reliably collecting a desired object without applying a load to the object. It is to be.

(1) A nanopipette according to the present invention includes a cylindrical pipette main body that extends along a central axis and has a first suction passage capable of sucking an object formed therein, and Co, Ni, Cr, and Mo. A pipette part formed of a Co-Ni base alloy and extending along the central axis and having a base end fixed to the pipette body, wherein the pipette part is capable of sucking an object. A suction passage is communicated with the first suction passage, and is formed in a cylindrical suction portion formed inside in a state where the suction passage is open to the tip portion, and is formed so as to protrude from the tip portion of the suction portion. And a protruding piece that can be elastically deformed in a radial direction perpendicular to the central axis.

  According to the nanopipette according to the present invention, the pipette portion is made of a Co—Ni base alloy that is high strength, high corrosion resistance, high heat resistance, non-magnetic material, and further has high elasticity. Therefore, from the viewpoint that the Co—Ni-based alloy is a highly elastic material, for example, by contacting the protruding piece against the substrate on which the object is placed, the protruding piece is elastic so as to bend in the radial direction. Can be deformed. Thereby, even if the objects are connected to each other, for example, the connection between the objects can be disconnected using the elastic deformation of the protrusion pieces, and the desired object can be separated from the surroundings. Therefore, a desired object can be sucked into the second suction passage and collected through the first suction passage.

In particular, the external force is not directly applied to the desired object, but the desired object is separated and separated from the surroundings using the protruding pieces, so that the load on the desired object can be suppressed. it can. Therefore, the object can be collected in a state where the load acting on the desired object is suppressed. Furthermore, since the projecting piece is formed in the suction portion, it is possible to collect the target object easily, reliably and promptly after separating the desired object from the surroundings.
Note that since the Co—Ni-based alloy is a highly elastic material, by releasing the pressing of the protruding piece, the protruding piece can be elastically restored and restored to its original state. Therefore, the protruding piece can be repeatedly elastically deformed, and the object can be collected stably and repeatedly. In addition, from the viewpoint that the Co—Ni-based alloy is a high-strength material, even if the protruding piece is strongly pressed against the substrate, problems such as deformation, cracks, and defects are unlikely to occur in the protruding piece. Therefore, it can be set as the high quality nanopipette which can be used over a long period of time.

(2) The suction portion may be gradually reduced in diameter from the proximal end portion toward the distal end portion.

  In this case, since it can be set as a tapered suction part, it is easy to grasp the target object, and it becomes easy to accurately approach the protruding piece to the target position. Accordingly, it is easy to separate the object using the protruding pieces.

(3) The protruding piece protrudes away from the tip portion along the central axis from the tip portion, and gradually protrudes radially from the center axis as the distance from the tip portion increases. In addition, it may be inclined outward.

  In this case, since the protruding piece is inclined outward, when the protruding piece is pressed against the substrate on which the object is placed, the protruding piece is elastically deformed so as to bend in the radial direction with less resistance. be able to. Thereby, the connection between the objects can be smoothly disconnected, and the desired object can be easily separated quickly.

(4) A plurality of the projecting pieces may be formed at intervals along a circumferential direction around the central axis.

  In this case, when the projecting piece is pressed, a plurality of projecting pieces can be arranged so as to surround a desired object. For this reason, the connection between the desired object and other surrounding objects can be easily separated over the entire circumference of the object by pressing the plurality of projecting pieces once. Accordingly, it is easy to separate a desired object more smoothly, and it is easy to quickly collect the object.

(5) The protruding piece may be formed with a circumferential groove extending along a circumferential direction around the central axis.

  In this case, since the circumferential groove is formed in the protruding piece, when the protruding piece is pressed, the protruding piece can be urged to deform with the circumferential groove as a base point. Therefore, the protruding piece can be elastically deformed so as to bend in the radial direction with less resistance, and the object can be easily separated.

  According to the nanopipette according to the present invention, a desired object can be easily and reliably collected without applying a load to the object.

1 is a perspective view showing an embodiment of a nanopipette according to the present invention. It is a side view of the pipette part periphery in the nanopipette shown in FIG. It is a block diagram which shows an example of the FIB processing apparatus used for preparation of the nanopipette shown in FIG. FIG. 2 is a process diagram for producing the nanopipette shown in FIG. 1, and is a perspective view showing a state where a pipette portion is created by FIB processing a block member of a Co—Ni based alloy. It is a perspective view which shows the state which has fixed the pipette part and the pipette main body using the deposition film | membrane. It is a side view which shows the state which has cut | disconnected the connection of a desired cell and another cell using the protrusion piece of the pipette part shown in FIG. It is a side view which shows the modification of the nanopipette shown in FIG. It is sectional drawing which shows the state which has cut | disconnected the connection of a desired cell and another cell using the protrusion piece of the pipette part shown in FIG. It is a side view which shows another modification of the nanopipette shown in FIG.

  Hereinafter, embodiments of a nanopipette according to the present invention will be described with reference to the drawings. In the present embodiment, a cell (cultured cell) will be described as an example of an object to be collected. In each drawing, the scale of each component is appropriately changed in order to make the drawing easy to understand and to help understand the invention.

  As shown in FIGS. 1 and 2, the nanopipette 1 of the present embodiment extends along a central axis O and has a cylindrical pipette body 2 in which a first suction passage 10 capable of sucking cells C is formed. And a pipette portion 3 formed of a Co—Ni based alloy containing Co, Ni, Cr, and Mo, and fixed to the tip portion 11 of the pipette body 2.

In a plan view viewed from the direction of the central axis O, a direction orthogonal to the central axis O is referred to as a radial direction, and a direction around the central axis O is referred to as a circumferential direction.
Further, FIG. 1 illustrates a case where a plurality of cells C are cultured in a state of being attached to the substrate 4. Examples of the substrate 4 include a bottom wall portion in a culture vessel (not shown). The plurality of cells C form a colony-like cell mass M that is closely connected to each other and aggregated as the culture grows.

  The pipette body 2 is an extremely thin glass tube having an inner diameter, that is, a diameter of the first suction passage 10 of, for example, several hundred nm to several hundred μm. However, the pipette main body 2 is not limited to the glass tube, and may be formed of other materials. Moreover, the pipette main body 2 should just be formed in the cylinder shape which has the 1st suction passage 10 inside, and is not limited to a cylindrical shape.

The pipette body 2 is held in a cantilevered manner by, for example, a manipulator arm of a manipulator device (not shown) whose base end portion 12 can be operated three-dimensionally. Thereby, the whole nanopipette 1 is finely and accurately three-dimensionally operated by the manipulator device.
However, the present invention is not limited to this case. For example, an operation glass tube having a larger diameter is connected to the base end portion 12 side of the pipette main body 2, and the operator holds the operation glass tube, and the microscope The nanopipette 1 may be manually operated while visually recognizing the above.

  The pipette body 2 can be equipped with a pressure adjustment mechanism 13 that adjusts the internal pressure in the first suction passage 10. Thus, the cell C can be sucked into the first suction passage 10 by making the pressure inside the first suction passage 10 negative by the pressure adjusting mechanism 13. On the other hand, by making the inside of the first suction passage 10 positive by the pressure adjusting mechanism 13, for example, various solutions supplied into the first suction passage 10 (for example, liquids necessary for culturing the cells C). Or a test solution or the like) can be applied to the cells C.

  The pipette portion 3 extends along the central axis O, and has a cylindrical suction portion 20 in which a base end portion 21 is fixed to the distal end portion 11 of the pipette body 2 and a second suction passage 22 is formed inside. And a protruding piece 25 that is formed so as to protrude further from the distal end portion 23 of the suction portion 20 toward the distal end side and is elastically deformable in the radial direction.

The pipette part 3 of the present embodiment is fixed to the tip part 11 of the pipette body 2 by a deposition film D described later by FIB irradiation. This will be described later.
However, the fixing of the pipette part 3 is not limited to the deposition film D, and may be fixed by other fixing methods such as adhesion and welding, for example, and may be fixed using fitting such as locking. It doesn't matter.

  The outer shape of the suction part 20 is formed in a conical shape whose diameter is gradually reduced from the base end part 21 fixed to the pipette main body 2 toward the distal end part 23 where the protruding piece 25 is formed. However, the outer shape of the suction portion 20 is not limited to a conical shape, and may be formed in, for example, a quadrangular pyramid shape, and has the same outer diameter from the proximal end portion 21 to the distal end portion 23. It may be formed in a cylindrical shape.

  The second suction passage 22 is a passage through which the cells C can be sucked, and communicates with the first suction passage 10 and opens to the distal end portion 23 side of the suction portion 20. In the illustrated example, the second suction passage 22 gradually decreases in diameter from the base end portion 21 toward the distal end portion 23 corresponding to the outer shape of the suction portion 20, and from the opening area on the first suction passage 10 side. Also, the opening area on the tip end portion 23 side is formed to be small. Note that the opening area on the distal end portion 23 side in the second suction passage 22 is sized to suck the cells C.

  The protruding piece 25 is formed integrally with the distal end portion 23 of the suction portion 20. However, the present invention is not limited to this case, and the protruding piece 25 may be formed separately from the suction unit 20 and fixed to the suction unit 20 using the deposition film D, for example.

  The projecting piece 25 protrudes from the distal end portion 23 of the suction portion 20 so as to be separated from the distal end portion 23 along the central axis O, and is gradually separated from the central axis O in the radial direction as the distance from the distal end portion 23 increases. So that it slopes outward.

In the illustrated example, the protruding piece 25 is formed such that the peripheral width along the circumferential direction is slightly longer at the tip end portion 27 which is a free end than the root portion 26 located on the suction portion 20 side. Yes. However, the shape of the protruding piece 25 is not limited to this case. For example, the protruding piece 25 may be formed so that the circumferential width of the tip portion 27 is the same as or shorter than the circumferential width of the root portion 26.
Further, in the illustrated example, the protruding piece 25 is formed so that the thickness gradually decreases from the root portion 26 toward the tip portion 27.

A specific composition of the metal material constituting the pipette part 3 constituted by the suction part 20 and the protrusion piece 25 described above, that is, a Co—Ni base alloy containing Co, Ni, Cr and Mo will be described.
As a Co-Ni base alloy, the composition is in mass ratio, Co: 28-42%, Cr: 10-27%, Mo: 3-12%, Ni: 15-40%, Ti: 0.1-1% , Mn: 1.5% or less, Fe: 0.1 to 26%, C: 0.1% or less, Nb: 3% or less, and preferably a composition composed of the remaining inevitable impurities.
More preferably, in addition to the above composition, at least one selected from the group consisting of W: 5% or less, Al: 0.5% or less, Zr: 0.1% or less, B: 0.01% or less is included. That is good. The reason for limiting these composition ranges will be briefly described below.

Co itself has a large work-hardening ability, has the effect of reducing notch brittleness, increasing fatigue strength, and increasing high-temperature strength, but the effect is weak at less than 28%, and the matrix becomes too hard when it exceeds 42%. The face-centered cubic lattice phase becomes unstable with respect to the close-packed hexagonal lattice phase. Therefore, the mass ratio of Co is preferably 28 to 42%.
Cr is an essential component to ensure corrosion resistance and has the effect of strengthening the matrix. However, if it is less than 10%, the effect of obtaining excellent corrosion resistance is weak, and if it exceeds 27%, the workability and toughness are drastically reduced. To do. Therefore, the mass ratio of Cr is preferably 10 to 27%.

Mo has the effect of strengthening by solid solution in the matrix, the effect of increasing the work hardening ability, and the effect of improving the corrosion resistance in the coexistence with Cr, but if less than 3%, the desired effect cannot be obtained, 12% If it exceeds 1, the workability will be drastically lowered and a brittle σ phase is likely to be formed. Therefore, the mass ratio of Mo is preferably 3 to 12%.
Ni stabilizes the face-centered cubic lattice phase, maintains the workability, and improves the corrosion resistance. However, in the composition range of Co, Cr, Mo, Nb, and Fe, Ni is less than 15% and stable face-centered cubic. It is difficult to obtain a lattice phase, and if it exceeds 40%, the mechanical strength decreases. Therefore, the mass ratio of Ni is preferably 15 to 40%.

Ti has strong deoxidation, denitrification, desulfurization effects, and ingot structure refinement effects, but the effect is weak at less than 0.1%, and inclusions increase in the alloy when it exceeds 1%. or η phase _(Ni 3 Ti) toughness precipitated is decreased. Therefore, the mass ratio of Ti is preferably 0.1 to 1%.
Mn has the effect of deoxidation and desulfurization and the effect of stabilizing the face-centered cubic lattice phase, but if it is too much, the corrosion resistance and oxidation resistance are deteriorated. Therefore, the mass ratio of Mn is preferably 1.5% or less.

When Fe is too much, the oxidation resistance decreases, but considering the importance of strengthening the solid solution by dissolving in the matrix rather than the oxidation resistance, the mass ratio is preferably 0.1 to 26%. .
C dissolves in the matrix and forms carbides with Cr, Mo, Nb, W, etc., and has the effect of preventing the coarsening of the crystal grains. However, if it is too much, the toughness is lowered and the corrosion resistance is deteriorated. Therefore, the mass ratio of C is preferably 0.1% or less.

Nb has a solid solution in the matrix and strengthens it, and has an effect of increasing work hardening ability. However, if it exceeds 3%, the σ phase and δ phase (Ni ₃ Nb) are precipitated and the toughness is lowered. When Nb is contained, the mass ratio is preferably 3% or less.
W has the effect of strengthening the solid solution in the matrix and remarkably increasing the work hardening ability. However, if it exceeds 5%, the σ phase is precipitated and the toughness is lowered. The mass ratio is preferably 5% or less.

Al has the effect of improving deoxidation and oxidation resistance. However, if it is too much, deterioration of corrosion resistance or the like occurs. Therefore, when Al is contained, the mass ratio is preferably 0.5% or less.
Zr has the effect of increasing the grain boundary strength at high temperature and improving the hot workability, but if too much, the workability deteriorates conversely, so when containing Zr, the mass ratio is 0.1. % Or less is preferable.
B has an effect of improving the hot workability, but if it is too much, conversely, the hot workability is lowered and the crack is liable to break. Therefore, when B is contained, the content is preferably 0.01% or less.

Furthermore, it is more preferable that the mass ratio of Fe is 0.1 to 3% and Nb: 3% or less is included.
That is, the composition is in mass ratio, Co: 28-42%, Cr: 10-27%, Mo: 3-12%, Ni: 15-40%, Ti: 0.1-1%, Mn: 1.5 A Co—Ni-based alloy composed of% or less, Fe: 0.1 to 3%, C: 0.1% or less, Nb: 3% or less, and the balance inevitable impurities is more preferable.
In the Co—Ni based alloy having such a composition, it is possible to more effectively prevent the oxidation resistance from being lowered by setting the upper limit of Fe to 3%.

  The Co—Ni-based alloy having the above composition has high strength, high corrosion resistance, high heat resistance, nonmagnetic properties, high elasticity, and excellent plastic workability. In particular, this Co-Ni-based alloy has high work hardening performance, and after being subjected to plastic deformation in the cold, it is subjected to strain age hardening by performing an aging heat treatment in a high temperature region (for example, 400 ° C to 800 ° C). It is an age-hardening type alloy that is strengthened.

In the present embodiment, as the cold working, the pipette part 3 is processed by FIB processing described later, and then aging heat treatment is performed. Thereby, the pipette part 3 is equipped with the characteristic of 80-85 GPa, for example as a Young's modulus (longitudinal elastic modulus) 200-240 GPa, and a rigidity (shear elastic modulus or transverse elastic modulus).
As described above, since the pipette portion 3 has a high Young's modulus and a high rigidity, in particular, the protruding piece 25 can be elastically deformed in the radial direction while having a high mechanical strength.

(Production of nanopipette)
Next, the case where the nanopipette 1 configured as described above is manufactured will be briefly described. Conventionally, there has been no disclosure or suggestion regarding the production of the nanopipette 1 by finely processing a difficult-to-cut material such as a Co—Ni-based alloy.
On the other hand, in the present embodiment, as shown in FIG. 3, by using the FIB processing apparatus 30 that can irradiate two charged particle beams of FIB (focused ion beam) and EB (electron beam), it is possible to accurately. Nanopipette 1 can be produced. Hereinafter, production of the nanopipette 1 will be described.

First, the FIB processing apparatus 30 will be briefly described.
The FIB processing apparatus 30 includes a first tweezers 32 capable of sandwiching a block member 31 made of a Co-Ni base alloy, a second tweezers 33 capable of sandwiching a pipette body 2, an irradiation mechanism 34 for irradiating FIB and EB, and an FIB. Alternatively, based on the detected secondary charged particles E, a detector 35 that detects secondary charged particles E generated by irradiation with EB, a gas gun 36 that supplies a compound gas G for forming the deposition film D, and the like. A control unit 38 for generating image data and displaying the image data on the display unit 37, and a vacuum chamber 39.

  For example, as shown in FIG. 4, the block member 31 has a length L slightly longer than the length along the central axis O from the base end portion 21 of the suction portion 20 to the distal end portion 27 of the protruding piece 25. It is a rectangular parallelepiped block of a Co—Ni base alloy formed in a size having a width W and a thickness T slightly longer than the outer diameter of the base end portion 21.

As shown in FIG. 3, the first tweezers 32 and the second tweezers 33 displace the tweezers 40 and the tweezers 40 in, for example, five axes (movements to the X axis and the Y axis that are parallel to the horizontal plane and orthogonal to each other, A displacement mechanism 41 that moves to a Z-axis orthogonal to the X-axis and the Y-axis, tilts around the X-axis or Y-axis, and rotates around the Z-axis), and is disposed in the vacuum chamber 39. Yes.
The first tweezers 32 holds the block member 31 stably by sandwiching the block member 31 with the tweezers 40. The second tweezers 33 hold the pipette body 2 stably by sandwiching the pipette body 2 with the tweezers 40.

  The irradiation mechanism 34 is disposed above the first tweezers 32 and the second tweezers 33. For example, the FIB barrel 45 that irradiates FIB parallel to the Z axis, and the SEM that irradiates EB obliquely with respect to the Z axis. A lens barrel 46.

The FIB column 45 includes an ion generation source 45a and an ion optical system 45b. After the ions generated from the ion generation source 45a are finely squeezed into an FIB by the ion optical system 45b, a block member is formed in the vacuum chamber 39. It is possible to irradiate the FIB 31 or the pipette body 2.
The SEM column 46 has an electron generation source 46a and an electron optical system 46b. After the electrons generated by the electron generation source 46a are finely squeezed into EB by the electron optical system 46b, a block member is formed in the vacuum chamber 39. It is possible to irradiate EB toward 31 and the pipette body 2.

The detector 35 detects secondary charged particles E such as secondary electrons and secondary ions generated from the block member 31 and the pipette main body 2 when FIB or EB is irradiated, and outputs the secondary charged particles E to the control unit 38. Yes. The gas gun 36 emits a compound gas (raw material gas) G containing a substance (for example, phenanthrene, platinum, carbon, tungsten, etc.) that is a raw material of the deposition film D.
The compound gas G is decomposed by the secondary charged particles E and separated into a gas component and a solid component. Then, the deposited solid component is deposited to form the deposition film D.

  The control unit 38 comprehensively controls each component described above, and controls the acceleration voltage and beam current of the FIB column 45 and the SEM column 46 to change. In particular, the control unit 38 can freely adjust the FIB beam diameter by changing the acceleration voltage and the beam amount of the FIB column 45. Thereby, it is possible not only to acquire an observation image but also to locally etch (FIB) the block member 31, for example.

The control unit 38 converts the secondary charged particles E detected by the detector 35 into, for example, a luminance signal to generate observation image data, and then displays the observation image on the display unit 37 based on the observation image data. It is output. Thereby, the display part 37 displays an observation image.
An input unit 47 that can be input by an operator is connected to the control unit 38. Thereby, the control unit 38 controls each component based on the signal input by the input unit 47. That is, the operator irradiates a desired region with FIB or EB through the input unit 47 for observation, irradiates the desired region with FIB, performs etching, or applies the compound gas G to the desired region. The deposition film D can be deposited by irradiating FIB while supplying.

Next, production of the pipette unit 3 using the FIB processing apparatus 30 will be described.
As shown in FIG. 4, after holding one end portion of the block member 31 with the first tweezers 32, the block member 31 is irradiated with FIB while appropriately changing the posture of the block member 31, so that a part of the block member 31 is cut off. The member 31 is continuously etched. Thereby, while being able to form the external shape of the suction part 20 and the protrusion piece 25, respectively, the suction part 20 and the protrusion piece 25 can be formed integrally. Furthermore, by simultaneously irradiating the suction part 20 with FIB simultaneously or before and after this, a hole can be made in the suction part 20 so as to penetrate the suction part 20 along the central axis O. . Thereby, the second suction passage 22 can be formed in the suction portion 20.

  As described above, the pipette unit 3 can be manufactured by cold working using the FIB processing apparatus 30. Next, an aging heat treatment is performed on the produced pipette part 3 to impart high elasticity to the pipette part 3 by strain age hardening.

After performing the heat treatment, again using the FIB processing apparatus 30, as shown in FIG. 5, the base end portion 21 of the pipette portion 3 held by the first tweezers 32 and the pipette main body held by the second tweezers 33 The two tip portions 11 are brought into contact with each other. Then, FIB is irradiated while supplying the compound gas G from the gas gun 36 around the contact portion between the pipette unit 3 and the pipette body 2.
Thereby, the secondary charged particle E generated by irradiating the contact portion between the pipette unit 3 and the pipette body 2 with the FIB decomposes the compound gas G and separates it into a gas component and a solid component. Then, the separated solid component is deposited on the contact portion between the pipette unit 3 and the pipette body 2 to form the deposition film D.

  As a result, the pipette part 3 and the pipette body 2 can be fixed to each other, and the nanopipette 1 shown in FIGS. 1 and 2 can be produced. Therefore, according to the above-described production method, the pipette part 3 can be produced with high accuracy and further efficiency even with a Co—Ni based alloy known as a difficult-to-cut material.

(Operation of nanopipette)
Next, a case where a desired cell C1 is collected from the cell mass M shown in FIG. 1 using the nanopipette 1 configured as described above will be described.

  According to the nanopipette 1 of the present embodiment, the pipette portion 3 is made of a Co—Ni-based alloy that is high strength, high corrosion resistance, high heat resistance, non-magnetic material, and further has high elasticity. Therefore, from the viewpoint that the Co—Ni-based alloy is a highly elastic material, as shown in FIG. 6, the protruding piece 25 is brought into contact with the substrate 4 to which the cells C are attached by pressing the protruding piece 25. 25 can be elastically deformed so as to bend in the radial direction.

  Thereby, even if the cells C are connected to each other to form the cell mass M, the connection between the cells C can be disconnected using the elastic deformation of the projection piece 25, and the desired cell C1 is separated from the surroundings. Can be made. That is, a desired cell C1 can be selected and separated from the cell mass M.

  At this time, the above-described detaching operation by the protruding piece 25 may be repeated according to the connection state between the desired cell C1 and another cell C. For example, when the desired cell C1 is connected to other cells C over the entire circumference, the separation operation is performed while changing the position of the projection piece 25 along the circumferential direction of the desired cell C1. Just do it. In FIG. 6, the state which has cut | disconnected the last connection part of the desired cell C1 and the other cell C is illustrated.

  Then, after separating the desired cell C1, the inside of the first suction passage 10 is set to a negative pressure by the pressure adjusting mechanism 13, thereby bringing the desired cell C1 into the second suction passage 22 as shown in FIG. Suction can be performed and sampled through the first suction passage 10.

  In particular, the external force is not directly applied to the desired cell C1, but the desired cell C1 is separated and separated from the surroundings using the protrusions 25, so that the load on the desired cell C1 is suppressed as much as possible. be able to. Therefore, the cell C1 can be collected in a state where the load acting on the desired cell C1 is suppressed, and the desired cell C1 can be collected alive.

  Furthermore, since the protrusion piece 25 is formed in the suction part 20, after separating the desired cell C1 from the surroundings, the cell C1 can be collected easily, reliably and quickly. Therefore, the cell C1 collected alive can be used promptly for any subsequent process.

Since the Co—Ni based alloy is a highly elastic material, after the desired cell C1 is collected, the nanopipette 1 is pulled up to release the pressing of the protruding piece 25, thereby elastically deforming the protruding piece 25. To restore the original state. Therefore, the protruding piece 25 can be elastically deformed repeatedly, and desired cells C1 can be collected stably and repeatedly.
Further, from the viewpoint that the Co—Ni-based alloy is a high-strength material, even if the protrusion piece 25 is strongly pressed against the substrate 4, defects such as deformation, cracks, and defects occur in the protrusion piece 25. hard. Therefore, it can be set as the high quality nanopipette 1 which can be used over a long period of time.

As described above, according to the nanopipette 1 of the present embodiment, a desired cell C1 can be selected and separated from the cell mass M, and easily and without applying a load to the cell C1. It can be reliably collected alive.
Moreover, since the protruding pieces 25 are inclined outward, when the protruding pieces 25 are pressed against the substrate 4, the protruding pieces 25 can be elastically deformed so as to bend in the radial direction with little resistance. Thereby, the connection between the cells C can be disconnected smoothly, and the desired cell C1 can be easily separated quickly.

  Furthermore, since the suction part 20 is gradually reduced in diameter from the proximal end part 21 toward the distal end part 23, the suction part 20 can be tapered. Therefore, the field of view is not easily obstructed by the suction unit 20, and it is easy to grasp the desired cell C1. Therefore, it becomes easy to accurately approach the projecting piece 25 to the target position, and the cell C1 can be easily separated using the projecting piece 25.

  As mentioned above, although embodiment of this invention was described, these embodiment is shown as an example and is not intending limiting the range of invention. The embodiment can be implemented in various other forms, and various omissions, replacements, and changes can be made without departing from the spirit of the invention. Further, the embodiments include, for example, those that can be easily assumed by those skilled in the art, substantially the same, and equivalents.

  In the said embodiment, although the cell C was mentioned as an example and demonstrated as a target object, it is not limited to the cell C. For example, a living tissue obtained from a human body, an animal, a plant, or the like may be used.

  Moreover, in the said embodiment, although only one protrusion piece 25 was formed, it is not limited to this case. For example, as shown in FIG. 7, a nanopipette 50 in which a plurality of protruding pieces 25 are formed at intervals along the circumferential direction may be used.

  According to the nanopipette 50 configured as described above, as shown in FIG. 8, when the protruding piece 25 is pressed against the substrate 4, a plurality of protruding pieces 25 can be arranged so as to surround a desired cell C1. . Therefore, the connection between the desired cell C1 and other surrounding cells C can be easily separated over the entire circumference of the desired cell C1 by pressing the plurality of protrusions 25 once. Therefore, the desired cell C1 can be more easily and accurately separated, and the cell C1 can be easily collected.

Furthermore, as shown in FIG. 9, a nanopipette 60 in which circumferential grooves 61 extending in the circumferential direction are formed in the plurality of projecting pieces 25 may be used.
In the example shown in the figure, two circumferential grooves 61 are formed with a spacing in the direction of the central axis O with respect to each projection piece 25. However, the number of the circumferential grooves 61 may be formed only one for each protrusion piece 25, or may be formed in three or more multistages in the direction of the central axis O.

According to the nanopipette 60 configured as described above, since the circumferential groove 61 is formed in the protruding piece 25, when the protruding piece 25 is pressed, the protruding piece 25 is urged to deform with the circumferential groove 61 as a base point. be able to. Therefore, the protruding piece 25 can be elastically deformed so as to bend in the radial direction with less resistance, and desired cells C1 can be easily separated.
In FIG. 9, the case where a plurality of protruding pieces 25 are provided is described as an example. However, in the nanopipette 1 having one protruding piece 25 shown in FIGS. The circumferential groove 61 may be formed.

In the present embodiment, the pipette portion 3 is made of a Co—Ni base alloy that has high strength, high corrosion resistance, high heat resistance, non-magnetic material, and is also highly elastic. The material of the part 3 is another metal material, and may be an elastically deformable metal material such as Al. The Co—Ni based alloy in this embodiment can be expected to have a remarkable effect as described above, but even a metal material that can be elastically deformed such as Al is expected to have a sufficient effect depending on the use environment. be able to.
That is, the pipette main body is formed of an elastically deformable metal material, and extends along the central axis. And a pipette portion having a base end portion fixed to the pipette body, wherein the pipette portion has a second suction passage capable of sucking an object communicating with the first suction passage and opening at a distal end portion. A cylindrical suction part formed inside in a state of being formed, and a protruding piece that is formed so as to protrude from the tip part of the suction part and elastically deformable in a radial direction perpendicular to the central axis A nanopipette may be provided.

C ... Cell (object)
O ... center axis 1, 50, 60 ... nanopipette 2 ... pipette body 3 ... pipette part 10 ... first suction passage 20 ... suction part 21 ... base end part of suction part 22 ... second suction path 23 ... tip of suction part Part 25 ... Projection piece 61 ... Circumferential groove

Claims (5)

A tubular pipette body extending along the central axis and having a first suction passage capable of sucking an object formed therein;
A pipette part formed of a Co-Ni base alloy containing Co, Ni, Cr and Mo, extending along the central axis and having a base end fixed to the pipette body,
The pipette part is
A cylindrical suction part formed inside with a second suction path capable of sucking an object communicating with the first suction path and opened at a tip part;
A nanopipette, comprising: a protruding piece that is formed so as to protrude from the distal end portion of the suction portion and elastically deformable in a radial direction perpendicular to the central axis. The nanopipette according to claim 1,
The said suction part is a nanopipette which is gradually diameter-reduced as it goes to the said front-end | tip part from the said base end part. The nanopipette according to claim 1 or 2,
The protruding piece protrudes from the distal end portion along the central axis so as to be separated from the distal end portion, and gradually increases away from the central axis as the distance from the distal end portion increases. A nanopipette that is inclined in the direction. The nanopipette according to any one of claims 1 to 3,
A plurality of the projecting pieces are formed at intervals along a circumferential direction that circulates around the central axis. The nanopipette according to any one of claims 1 to 4,
The nanopipette, wherein the protruding piece is formed with a circumferential groove extending along a circumferential direction around the central axis.

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