JP6740071B2 - knife - Google Patents

Description

The present invention relates to knives.

Generally, in the field of biology, in order to inspect and study a biological sample obtained from a human body, an animal, a plant or the like, for example, a biological tissue or a cell, a target such as a biological sample using a fine and sharp knife. You may cut things.
As this kind of knife, a metal knife having a sharpened cutting edge by mechanical processing such as polishing is known. However, since the metal knife only has a sharpened cutting edge by machining, there is a limit in machining even if it is sharpened as much as possible, and the thickness of the cutting edge is likely to be about 1 mm. Therefore, when a cell having a diameter of several tens of μm to several hundreds of μm is cut by using a metal knife, the cutting is likely to be performed while the cell is being crushed by a cutting edge, which gives stress to the cell.

However, it is not preferable to give stress to cells. For example, it is desired to reduce the stress applied to cells in artificial insemination, cloning technology, and inspections and researches in the field of regenerative medicine. It is difficult for a conventional metal knife to meet such a demand.

Then, as a knife thinner than a metal knife, for example, a micro knife having a blade made of a solid thin film having a thickness of 10 μm (see Patent Document 1) or a micro knife having a cutting blade having a thickness of 30 μm (see Patent Document 2). )It has been known.
Furthermore, as other knives for cutting a biological sample, for example, a vibrating knife that vibrates in a direction different from the cutting direction (see Patent Document 3) and a micro knife that vibrates in the cutting direction (see Patent Document 4) are known. ..

JP-A-11-138494 JP, 2008-286528, A JP 2004-305441 A JP, 2010-261757, A

By the way, since the biological sample, which is represented by cells, has flexibility, the biological sample is likely to be deformed so as to be partially recessed following the pressing of the knife during cutting with a knife. Therefore, even when the biological sample is cut using the conventional thin microknife, the biological sample is pressed against the substrate (for example, the slide plate or the bottom plate of the petri dish) via the blade edge. Sometimes, there is no choice but to disconnect.

However, in this case, for example, when the blade tip was inserted into the biological sample, it was easy for the blade tip to come into contact with the substrate so as to strongly press it. Therefore, there is a possibility that the cutting edge may be deformed, cracked or broken, and there is room for improvement. In particular, the thinner the knife, the more likely it is that the blade edge will become defective.

On the other hand, in the case of a knife that cuts a biological sample by utilizing the above-mentioned conventional vibration, the vibration is transmitted to the biological sample at the time of cutting, so that stress is easily applied.

The present invention has been made in view of such circumstances, and an object thereof is to cut an object while protecting the cutting edge portion and suppressing stress applied to the object such as a biological sample. Is to provide a knife.

(1) A knife according to the present invention is formed of a Co—Ni based alloy containing Co, Ni, Cr and Mo, and extends in one direction from a knife base end supported in a cantilever manner to the knife tip. An existing knife portion, and the knife portion is formed over the entire length with a cutting edge portion that comes into contact with an object, and a ridge portion that is arranged so as to face the cutting edge portion. Is 30 μm or less, and the knife portion is formed such that the knife thickness gradually decreases from the peak portion toward the blade edge portion.

According to the knife of the present invention, since the knife portion is formed such that the knife thickness (thickness) gradually decreases from the crest portion toward the blade tip portion, the blade tip portion formed over the entire length of the knife portion. Can be sharpened. Moreover, since the knife thickness in the peak portion is set to 30 μm or less, the knife thickness in the cutting edge portion can be further reduced, for example, an ultrathin thickness of several hundred nm to several μm. Thereby, an object such as a cell having a diameter of several μm to several hundreds μm can be cut without being crushed. Therefore, the object can be cut while suppressing the stress applied to the object.

In particular, the knife portion is formed of a Co—Ni based alloy which has high strength, high corrosion resistance, high heat resistance, non-magnetic property and high elasticity.
Therefore, from the viewpoint that the Co-Ni-based alloy is a high-strength material, when cutting the object, the blade tip of the knife portion makes contact with, for example, the substrate on which the object is placed so as to strongly press it. Even if it does, defects such as deformation, cracks, and defects are unlikely to occur in the ultrathin blade edge portion.

Moreover, from the viewpoint that the Co-Ni-based alloy is a highly elastic material, when the blade edge of the knife portion comes into contact with the support plate, the entire knife portion supported in a cantilevered manner is reversed in the knife width direction. Can be elastically deformed (curved). Therefore, it is possible to absorb the reaction force transmitted from the substrate to the blade portion when the blade portion comes into contact with the substrate. Therefore, it is possible to effectively suppress the occurrence of the above-mentioned problem in the extremely thin blade edge portion. As a result, the object can be cut while protecting the cutting edge.
By separating the blade tip from the substrate, the knife portion is elastically restored and deformed to return to the original state.

(2) The knife thickness at the cutting edge may be 100 nm or less.

(3) The knife length of the knife portion is 50 μm or less, the knife width at the knife base end portion is 20 μm or less, the knife thickness at the ridge portion is 10 μm or less, and the knife thickness at the cutting edge portion is 100 nm or less. May be

In this case, the blade edge portion can be made to enter the object with less resistance, and the contact area of the blade edge portion with respect to the object can be suppressed, so that the object is cut while further suppressing the stress applied to the object. be able to. Further, since it is difficult for the target object to adhere to the blade tip portion during cutting, it is possible to prevent the cut object from being dragged while being attached to the blade tip portion. This also makes it difficult to apply stress to the object.

(4) The knife portion may be formed such that the knife thickness and the knife width from the crest portion to the cutting edge portion are gradually reduced from the knife base end portion toward the knife tip end portion. ..

In this case, the shape of the knife portion can be a triangular pyramid shape with the cutting edge portion facing the object. Therefore, the cross-sectional area of the knife portion orthogonal to the extending direction of the knife portion is the largest on the side of the cantilever-supported knife base end, and gradually decreases toward the knife tip that is the free end. Thereby, the knife portion can be more easily elastically deformed, and even when the blade tip portion of the knife portion is strongly pressed against the substrate, the blade tip portion is less likely to cause a defect.

(5) The knife portion is located on the side of the knife tip portion, and the tip portion is inclined with respect to the ridge portion at a first inclination angle, and between the tip portion and the knife base end portion. And a knife main body in which the cutting edge portion is inclined at a second inclination angle with respect to the crest portion, the first main inclination angle being greater than the second inclination angle. May be larger.

In this case, the angle of inclination of the cutting edge with respect to the ridge can be increased at the tip on the knife tip side. Therefore, for example, while obliquely inclining the knife portion with respect to the substrate, when cutting the object mainly using the blade tip portion of the knife tip portion side, rather than pushing the blade tip portion of the pointed portion against the substrate, It can be arranged so as to approach a parallel state. Therefore, it is possible to protect the tip end portion (cutting edge) of the knife portion, which is likely to be deformed or chipped.

(6) The peak portion may be formed with a reflection surface that reflects the detection light emitted from the outside and faces different directions depending on the deformation of the knife portion.

In this case, since the reflecting surface faces different directions depending on the elastic deformation of the knife portion, the elastically deformed state of the knife portion can be grasped based on the detection light reflected by the reflecting surface. Therefore, the object can be cut while grasping the state of the knife portion more accurately.

(7) A holding member is formed so as to extend in the extending direction of the knife portion from the base end portion toward the tip end portion, and the base end portion is held in a cantilevered manner. The knife base end part may be attached to the holding member, and the holding member may be formed of a transparent material.

In this case, the knife member can be held more stably by using the holding member, so that the object can be cut stably. Further, since the knife portion can be operated by holding the holding member with, for example, a manipulator device, a finer and more accurate cutting operation can be performed. Moreover, since the holding member is transparent, the knife portion can be confirmed without being affected by the holding member. Therefore, the knife portion can be accurately approached to a desired position of the object.

According to the knife of the present invention, it is possible to protect the cutting edge portion and cut the object while suppressing the stress applied to the object such as the biological sample.

It is a perspective view which shows 1st Embodiment of the knife which concerns on this invention. It is the top view which expanded the knife front-end|tip part shown in FIG. FIG. 3 is a side view of the periphery of the knife tip portion shown in FIG. 2. It is a block diagram which shows an example of the FIB processing apparatus used for manufacture of the knife shown in FIG. FIG. 3 is a step diagram for manufacturing the knife shown in FIG. 1, and is a perspective view showing a state in which a part of a plate member of Co—Ni based alloy is subjected to FIB processing. FIG. 6 is a perspective view showing a state in which the plate member is further subjected to FIB processing from the state shown in FIG. 5. FIG. 7 is a perspective view showing a state where the plate member is further subjected to FIB processing from the state shown in FIG. 6 to form a triangular pyramid shape. FIG. 8 is a perspective view showing a state in which the tip portion of a portion formed in a triangular pyramid shape is further subjected to FIB processing and cut obliquely from the state shown in FIG. 7. FIG. 9 is a top view of the periphery of the triangular pyramidal tip portion shown in FIG. It is a side view of the triangular pyramid-shaped tip part periphery shown in FIG. It is a perspective view showing the state where the knife part and the holding member are fixed using a deposition film. It is a side view which shows the state which is cutting the cell using the knife shown in FIG. It is the side view which expanded the knife front-end|tip part shown in FIG. It is sectional drawing of the knife part along the AA line shown in FIG. It is a figure which shows 2nd Embodiment of the knife which concerns on this invention, Comprising: It is a block diagram of the manipulator system provided with a knife. It is a perspective view which shows the modification of the knife shown in FIG. It is a perspective view which shows another modification of the knife shown in FIG.

(First embodiment)
Hereinafter, a first embodiment of a knife according to the present invention will be described with reference to the drawings. In the present embodiment, a biological sample (cell) will be described as an example of an object to be cut. That is, in the present embodiment, the knife is used as the biological sample knife. In addition, in each drawing, the scale of each component is appropriately changed in order to make the drawing easy to see and to help understanding of the invention.

As shown in FIGS. 1 to 3, the biological sample knife 1 of the present embodiment has a knife portion 2 formed of a Co—Ni based alloy containing Co, Ni, Cr and Mo, and a knife portion 2 attached thereto. And a holding member 3.
The knife portion 2 is formed so as to extend in one direction along the knife axis O from the knife base end portion 10 toward the knife tip end portion 11, and the tip end portion 3 a of the holding member 3 via the knife base end portion 10. Is attached to. Thereby, the knife portion 2 is fixed to the holding member 3 in a state where the knife base end portion 10 is supported in a cantilever manner so that the knife tip end portion 11 becomes a free end.

The knife portion 2 is fixed to the tip portion 3a of the holding member 3 by a deposition film D generated by decomposition of a compound gas (raw material gas) G described later by irradiation of FIB. This will be described later.
However, the fixing of the knife portion 2 is not limited to the deposition film D, and may be fixed by another fixing method such as adhesion or welding, or may be fixed by using fitting such as locking. It doesn't matter.

In the knife portion 2, a cutting edge portion 12 that comes into contact with the cells C, and a ridge portion (a back portion of the knife portion 2) 13 arranged so as to face the cutting edge portion 12 are formed over the entire length. ing.
In the present embodiment, the direction from the ridge 13 to the cutting edge 12 is called downward, and the opposite direction is called upward. Therefore, the cutting edge portion 12 faces downward, and the ridge portion 13 faces upward. The length along the knife axis O is referred to as a knife length L, and the length along the up-down direction is referred to as a knife width (knife height) W. The length along the left-right direction is orthogonal to the knife axis O and the up-down direction. The length is called knife thickness T.

The knife portion 2 is formed so that the knife thickness T gradually becomes thinner from the crest portion 13 toward the cutting edge portion 12 over the entire length thereof. As a result, the cutting edge portion 12 is a sharp edge that continuously extends linearly from the tip (cutting edge) P of the knife portion 2 to the knife base end portion 10.

Further, the knife portion 2 is formed such that the knife thickness T and the knife width W from the crest portion 13 to the cutting edge portion 12 become gradually smaller as going from the knife base end portion 10 to the knife tip end portion 11. As a result, the knife portion 2 extends from the knife base end portion 10 toward the knife tip end portion 11 and is formed in a triangular pyramid shape with the blade tip portion 12 facing the cell C side.
Therefore, the cross-sectional area orthogonal to the knife axis O (extending direction of the knife portion 2) is the largest on the side of the knife base end portion 10 supported in a cantilever manner, and gradually increases toward the knife tip portion 11 which is the free end. It is getting smaller.

Further, the knife portion 2 is located on the side of the knife tip portion 11 and is continuously provided to the tip portion 14 in a state of being located between the tip portion 14 connected to the tip P and the tip portion 14 and the knife base portion 10. And a knife body 15. The tip portion 14 and the knife main body portion 15 have different inclination angles of the cutting edge portion 12 with respect to the ridge portion 13.

Specifically, as shown in FIG. 3, at the pointed portion 14, the cutting edge portion 12 is inclined with respect to the ridge portion 13 at a first inclination angle θ1. In the knife body 15, the cutting edge 12 is formed at a second inclination angle θ2 with respect to the crest 13. The first tilt angle θ1 is slightly larger than the second tilt angle θ2.
As described above, in the knife portion 2, the inclination angle of the cutting edge portion 12 with respect to the crest portion 13 is not constant from the knife base end portion 10 to the tip P, and the inclination angle is slightly small at the tip 14 near the tip P. It is formed to be large.

The knife portion 2 has one edge (side edge portion) and the other edge (side edge portion) of the ridge portion 13 from the knife base end portion 10 to the most distal end P in relation to the above-described inclination angle. The angle formed by is not constant, and is formed to be slightly larger at the tip portion 14.
Specifically, in the plan view of the ridge portion 13 as shown in FIG. 2, in the knife main body portion 15, an angle formed by one edge of the ridge portion 13 and the other edge is an angle θ3, and at the tip portion 14. Is an angle θ4 which is larger than the angle θ3 with respect to one edge of the ridge 13 and the other edge.
As described above, by forming the tip portion 14 connected to the tip P on the knife tip portion 11 side, as compared with the case where the tip portion 14 is not formed (that is, the shape shown by the dotted line in FIGS. 2 and 3). Thus, the durability of the knife portion 2 can be improved.

However, the shape of the knife portion 2 is not limited to the above-described case, and may be, for example, a shape in which the tip portion 14 is not formed. In particular, if the object can be cut using only the cutting edge portion 12, the shape without the pointed portion 14 may be used. In this case, the number of processing steps of the knife portion 2 can be reduced, so that the manufacturing cost can be reduced, for example.

A specific size of the knife unit 2 configured as described above will be described.
The knife length L is, for example, 200 μm or less, preferably 50 μm or less. The knife width W at the knife base 10 is, for example, 100 μm or less, and preferably 20 μm. The knife thickness T in the ridge 13 is, for example, 30 μm or less, and preferably 10 μm or less. The knife thickness T in the blade portion 12 is, for example, 100 nm or less.

Further, a specific composition of the metal material forming the knife portion 2, that is, a Co—Ni based alloy containing Co, Ni, Cr and Mo will be described.
As a Co-Ni based alloy, the composition is a mass ratio of Co: 28 to 42%, Cr: 10 to 27%, Mo: 3 to 12%, Ni: 15 to 40%, Ti: 0.1 to 1%. , Mn: 1.5% or less, Fe: 0.1 to 26%, C: 0.1% or less, Nb: 3% or less, and the composition having the balance unavoidable impurities is preferable.
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 good. The reason why these composition ranges are limited 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 if it is less than 28%, its effect is weak, and if it exceeds 42%, the matrix becomes too hard. Processing becomes difficult and 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 for ensuring the corrosion resistance, and also has the effect of strengthening the matrix, but if it is less than 10%, the effect of obtaining excellent corrosion resistance is weak, and if it exceeds 27%, the workability and toughness drop sharply. To do. Therefore, the mass ratio of Cr is preferably 10 to 27%.

Mo has the effect of forming a solid solution in the matrix to strengthen it, the effect of increasing work hardening ability, and the effect of enhancing corrosion resistance in the coexistence with Cr, but if it is less than 3%, the desired effect cannot be obtained, and if it is 12%, If it exceeds, the workability is sharply reduced and a brittle σ phase is easily generated. Therefore, the mass ratio of Mo is preferably 3 to 12%.
Ni stabilizes the face-centered cubic lattice phase, maintains the workability, and enhances the corrosion resistance, but in the composition range of Co, Cr, Mo, Nb, and Fe, when Ni is less than 15%, the face-centered cubic is stable. 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 a strong effect of deoxidizing, denitrifying, desulfurizing, and refining the ingot structure, but if it is less than 0.1%, its effect is weak, and if it exceeds 1%, inclusions increase in the alloy. Alternatively, the η phase (Ni ₃ Ti) precipitates and the toughness decreases. 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, it deteriorates corrosion resistance and oxidation resistance. Therefore, the mass ratio of Mn is preferably 1.5% or less.

If Fe is too much, the oxidation resistance decreases, but considering that the effect of solid solution in the matrix and strengthening this is emphasized rather than the oxidation resistance, the mass ratio is preferably 0.1 to 26%. ..
C forms a solid solution in the matrix and forms carbides with Cr, Mo, Nb, W, etc., and has the effect of preventing coarsening of crystal grains, but if it is too large, toughness decreases, corrosion resistance deteriorates, and the like. Therefore, the mass ratio of C is preferably 0.1% or less.

Nb has the effect of forming a solid solution in the matrix to strengthen it and increase the work hardening ability, but if it exceeds 3%, the σ phase or δ phase (Ni ₃ Nb) precipitates and the toughness decreases, When Nb is contained, the mass ratio is preferably 3% or less.
W has the effect of forming a solid solution in the matrix to strengthen it and significantly increase the work hardening ability, but if it exceeds 5%, the σ phase precipitates and the toughness decreases, so when W is contained, The mass ratio is preferably 5% or less.

Al has an effect of improving deoxidation and oxidation resistance, but if it is too much, corrosion resistance deteriorates. 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 hot workability, but if too much, the workability deteriorates. Therefore, when Zr is included, the mass ratio is 0.1. % Or less is preferable.
B has the effect of improving the hot workability, but if it is too much, the hot workability decreases and cracks easily. Therefore, when B is contained, it is preferably 0.01% or less.

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

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

In this embodiment, as the cold working, the knife portion 2 is worked by the FIB working described later, and then the aging heat treatment is performed. As a result, the knife portion 2 has, for example, a Young's modulus (longitudinal elastic modulus) of 200 to 240 Gpa and a rigidity modulus (shear elastic modulus or lateral elastic modulus) of 80 to 85 GPa.
As described above, since the knife portion 2 has high Young's modulus and rigidity, it can be elastically deformed in the knife width W direction (vertical direction) while having high mechanical strength in the knife axis O direction. It is said that.

As shown in FIG. 1, the holding member 3 is formed to extend along the knife axis O from the base end portion 3b toward the tip end portion 3a. In the illustrated example, the holding member 3 is formed in a transparent glass columnar shape. However, the holding member 3 is not limited to the columnar shape and is not limited to glass.

The holding member 3 is operated three-dimensionally, for example, by holding the base end portion 3b in a cantilevered manner by, for example, a manipulator arm or the like. Then, the knife base end portion 10 of the knife portion 2 is fixed to the tip end portion 3a of the holding member 3 via the deposition film D as described above.

(Preparation of knife for biological sample)
Next, a case of manufacturing the biological sample knife 1 configured as described above will be briefly described.
In the present embodiment, as shown in FIG. 4, a case where a FIB processing apparatus 20 capable of irradiating two charged particle beams of FIB (focused ion beam) and EB (electron beam) is used will be described as an example.

First, the FIB processing device 20 will be briefly described.
The FIB processing apparatus 20 includes a first tweezers 22 that can sandwich a plate member 21 made of a Co—Ni based alloy, a second tweezers 23 that can sandwich the holding member 3, an irradiation mechanism 24 that irradiates FIB and EB, and an FIB. Alternatively, based on the detector 25 that detects the secondary charged particles E generated by the irradiation of EB, the gas gun 26 that supplies the compound gas G for forming the deposition film D, and the detected secondary charged particles E A vacuum chamber 29 is provided with a control unit 28 that generates image data by using the control unit 28 and causes the display unit 27 to display the image data.

The plate member 21 is a Co—Ni formed in a size corresponding to the knife thickness T of the ridge portion 13, a length exceeding the knife length L, and a width corresponding to the knife width W of the knife base end portion 10. It is a base alloy flat plate member.

The first tweezers 22 and the second tweezers 23 displace the tweezers 30 and the tweezers 30 on, for example, 5 axes (movement to the X axis and the Y axis which are parallel to the horizontal plane and orthogonal to each other, orthogonal to the X axis and the Y axis). And a displacement mechanism 31 for rotating the Z-axis, tilting about the X-axis or Y-axis, and rotating about the Z-axis), and arranged in the vacuum chamber 29.
The first tweezers 22 stably holds the plate member 21 by sandwiching the plate member 21 with the tweezers portion 30. The second tweezers 23 stably hold the holding member 3 by sandwiching the holding member 3 with the tweezers portion 30.

The irradiation mechanism 24 is disposed above the first tweezers 22 and the second tweezers 23. For example, the FIB barrel 35 that irradiates the FIB parallel to the Z axis and the SEM that irradiates the EB obliquely to the Z axis. And a lens barrel 36.
The FIB lens barrel 35 has an ion generation source 35a and an ion optical system 35b. After the ions generated by the ion generation source 35a are narrowed down by the ion optical system 35b into FIB, a plate member is formed in the vacuum chamber 29. It is possible to irradiate 21 and the holding member 3 with FIB.
The SEM lens barrel 36 has an electron generation source 36a and an electron optical system 36b. After the electrons generated by the electron generation source 36a are narrowed down by the electron optical system 36b to be EB, a plate member is formed in the vacuum chamber 29. It is possible to irradiate 21 and the holding member 3 with EB.

The detector 25 detects the secondary charged particles E such as secondary electrons and secondary ions generated from the plate member 21 and the holding member 3 when irradiated with FIB or EB, and outputs them to the control unit 28. There is. The gas gun 26 releases 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 separated solid component is deposited to form the deposition film D.

The control unit 28 comprehensively controls each of the above-described components, and also controls to change the acceleration voltage and the beam current of the FIB lens barrel 35 and the SEM lens barrel 36. In particular, the control unit 28 can freely adjust the beam diameter of the FIB by changing the acceleration voltage and the beam amount of the FIB barrel 35. This makes it possible not only to obtain an observation image, but also to locally etch the plate member 21 (FIB process), for example.

In addition, the control unit 28 converts the secondary charged particles E detected by the detector 25 into, for example, a luminance signal to generate observation image data, and then displays the observation image on the display unit 27 based on the observation image data. It is outputting. As a result, the display unit 27 displays the observation image.
The control unit 28 is connected to an input unit 37 that allows an operator to input. As a result, the control unit 28 controls each component based on the signal input by the input unit 37. That is, the operator irradiates the desired area with FIB or EB for observation through the input unit 37, irradiates the desired area with the FIB for etching, and the desired area is exposed to the compound gas G. The deposition film D can be deposited by irradiating FIB while supplying the above.

Next, the production of the biological sample knife 1 using the FIB processing apparatus 20 will be described.
As shown in FIG. 5, after holding one end of the plate member 21 with the first tweezers 22, FIB is irradiated while appropriately changing the posture of the plate member 21, and the plate thickness gradually increases from the upper surface to the lower surface. The plate member 21 is continuously etched so as to be thin. Thereby, the ridge portion 13 and the cutting edge portion 12 can be formed on the plate member 21.
The one end of the plate member 21 held by the first tweezers 22 functions as a base 21a that does not irradiate the FIB.

Next, FIB is further irradiated while changing the posture of the plate member 21, and as shown in FIG. 6, the plate thickness decreases from the base portion 21a side of the plate member 21 toward the other end portion side of the plate member 21. Etching is continuously performed so that the thickness becomes gradually thinner.
Next, FIB is further irradiated while changing the posture of the plate member 21, and as shown in FIG. 7, the plate thickness decreases from the base portion 21a side of the plate member 21 toward the other end side of the plate member 21. Etching is continuously performed so as to become gradually smaller. Thereby, it can be processed into a triangular pyramid shape.

Then, as shown in FIG. 8, the tip portion of the triangular pyramid-shaped portion is irradiated with FIB, and the tip portion is viewed along the cutting plane F shown in FIG. Etching is done so that it is cut diagonally. As a result, as shown in FIGS. 9 and 10, it is possible to form the pointed portion 14 in which the cutting edge portion 12 is inclined with respect to the ridge portion 13 at the first inclination angle θ1.

As described above, the knife thickness T gradually decreases from the peak 13 toward the cutting edge 12, and the knife thickness T and the knife width W gradually decrease from the knife base end 10 toward the knife tip 11. The knife portion 2 thus formed can be formed in a state of being connected to the base portion 21a.

Next, the connecting portion between the knife portion 2 and the base portion 21a is irradiated with FIB to cut the connecting portion, and the knife portion 2 is separated from the base portion 21a. As a result, the knife portion 2 can be manufactured by cold working using the FIB processing device 20.
Next, the manufactured knife portion 2 is subjected to an aging heat treatment to impart high elasticity characteristics to the knife portion 2 by strain age hardening.

After performing the heat treatment, the FIB processing device 20 is used again, and as shown in FIG. 11, the knife portion 2 held by the first tweezers 22 and the tip portion 3a of the holding member 3 held by the second tweezers 23. Contact with. Then, the FIB is irradiated while supplying the compound gas G from the gas gun 26 around the contact portion between the knife portion 2 and the holding member 3. As a result, the secondary charged particles E generated by irradiating the contact portion between the knife portion 2 and the holding member 3 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 knife portion 2 and the holding member 3 to form the deposition film D.

As a result, as shown in FIG. 11, the knife portion 2 and the holding member 3 can be fixed to each other, and the biological sample knife 1 can be manufactured.

(Action of knife for biological sample)
Next, a case where the cells C are cut by using the biological sample knife 1 configured as described above will be described.

Since the knife portion 2 of the biological sample knife 1 of the present embodiment is formed so that the knife thickness T gradually decreases as the knife portion 2 moves from the crest portion 13 toward the cutting edge portion 12, it is formed over the entire length of the knife portion 2. The sharpened cutting edge portion 12 can be sharpened. Moreover, since the knife thickness T in the ridge portion 13 is set to 30 μm or less, the knife thickness T in the cutting edge portion 12 can be further reduced, for example, to be extremely thin, for example, about 100 nm.
Therefore, as shown in FIGS. 12 to 14, the cell C can be cut while suppressing the stress applied to the cell C.

In particular, as shown in FIG. 14, by setting the knife thickness T in the cutting edge portion 12 to 100 nm or less, the cutting edge portion 12 can enter the cell C with less resistance, and the cutting edge portion 12 contacts the cell C. Since the area can be suppressed, the cells C can be cut with good sharpness while further suppressing the stress applied to the cells C.
Moreover, since the cells C are less likely to adhere to the blade tip portion 12 during cutting, it is possible to prevent the cut cells C from being dragged while being attached to the blade tip portion 12. This also makes it difficult to apply stress to the cells C.

Further, since the knife portion 2 is stably supported in a cantilevered state by the holding member 3, the cells C can be stably cut. Moreover, since the holding member 3 is transparent, the knife portion 2 can be confirmed without being affected by the holding member 3. Therefore, the knife portion 2 can be accurately approached to a desired position of the cell C, and the cell C can be equally divided into two, for example.

In particular, the knife portion 2 is formed of a Co—Ni based alloy that has high strength, high corrosion resistance, high heat resistance, non-magnetic properties, and high elasticity.
Therefore, from the viewpoint that the Co—Ni-based alloy is a high-strength material, when the cells C are cut, the cutting edge portion 12 of the knife portion 2 strongly presses against the substrate 38 on which the cells C are placed, for example. Even with such contact, the defects such as deformation, cracks, and defects are unlikely to occur in the ultrathin blade edge portion 12.

Moreover, from the viewpoint that the Co—Ni based alloy is a highly elastic material, as shown in FIGS. 12 and 13, when the blade tip portion 12 of the knife portion 2 contacts the substrate 38, it is supported in a cantilevered manner. The entire knife portion 2 elastically deforms so as to warp (curve) in the knife width W direction (vertical direction).
Therefore, when the blade tip 12 contacts the substrate 38, the reaction force transmitted from the substrate 38 to the blade tip 12 can be absorbed. Therefore, it is possible to suppress the above-mentioned problems from occurring in the extremely thin blade tip 12. In particular, since the knife portion 2 is formed in the shape of a triangular pyramid, it is easily elastically deformed, and it is easy to effectively suppress the occurrence of defects in the cutting edge portion 12. As a result, the cells C can be cut while protecting the blade portion 12.

As described above, according to the biological sample knife 1 of the present embodiment, the cutting edge portion 12 can be protected, and the stress applied to the cells C can be suppressed, and the cells C can be cut with good resistance and good sharpness. You can

Further, in the knife portion 2, the angle of inclination of the cutting edge portion 12 with respect to the crest portion 13 is formed so that the tip portion 14 is larger than the knife body portion 15. Therefore, when the cells C are cut mainly by using the cutting edge 12 on the side of the knife tip 11 while inclining the knife 2 with respect to the board 38, as shown in FIG. Instead of pushing up the cutting edge portion 12 of the portion 14, the cutting edge portion 12 can be arranged so as to be parallel to the substrate 38.
Therefore, it is possible to protect the leading edge (cutting edge) P of the knife portion 2 which is apt to be deformed or chipped.

(Second embodiment)
Next, a second embodiment of the knife according to the present invention will be described with reference to the drawings. In the second embodiment, the same parts as those in the first embodiment are designated by the same reference numerals and the description thereof will be omitted.
In the present embodiment, a case where a manipulator system is used to operate the biological sample knife in the first embodiment to cut cells will be described as an example.

As shown in FIG. 15, the manipulator system 40 of the present embodiment includes a biological sample knife 1, a stage 42 that supports a cell 41 on which cells C are placed, and a manipulator arm 43 that operates the biological sample knife 1. And an optical microscope unit 44 for observing the cells C, and a control unit 45 such as a CPU for comprehensively controlling each of these components.

The cell 41 is formed of an optically transparent material in a dish shape having an upper opening, and is supported on the stage 42. A plurality of cells C, for example, are placed on the bottom surface of the cell 41 in a cultured state.
The stage 42 is arranged horizontally. In this embodiment, the direction parallel to the upper surface of the stage 42 is called the XY direction, and the direction perpendicular to the upper surface of the stage 42 is called the Z direction.

A through hole 42a is formed in the central portion of the stage 42 so as to penetrate the stage 42 in the Z direction. The cell 41 is supported on the stage 42 so as to close the through hole 42a from above. The stage 42 is movable in three directions of XY direction and Z direction.

The manipulator arm 43 is a multi-joint arm having a pair of hand portions 43a at its tip, and can be three-dimensionally curved in the XY direction and the Z direction by the drive unit 46.
Specifically, the manipulator arm 43 includes a plurality of joint portions 50 and a connecting portion 51 that connects the joint portions 50 to each other, and can be three-dimensionally curved as the joint portions 50 rotate. It is possible.

The joint portion 50 is, for example, a roll joint that rotates around a roll axis along the arm axis O1, or a yaw joint (or pitch joint) that rotates around a yaw axis (or pitch axis) orthogonal to the arm axis O1. There is. Then, these roll joints and yaw joints (or pitch joints) are connected so as to be arranged in an arbitrary arrangement (for example, alternately arranged). As a result, the manipulator arm 43 can be bent three-dimensionally.

In addition, each joint 50 can be rotated, for example, by an operation using an angle wire (not shown). The angle wire extends to the drive unit 46 through the inside of the manipulator arm 43.
The pair of hand parts 43a are configured to be movable toward and away from each other by an operation with a drive wire, for example. The drive wire extends to the drive unit 46 through the inside of the manipulator arm 43.

The drive unit 46 transmits the power to the pair of hand parts 43a via the drive wire to operate the pair of hand parts 43a, and the power to the respective joint parts 50 via the angle wires. And a joint actuator 53 that rotates the joint portion 50.
The hand actuator 52 and the joint actuator 53 operate based on an instruction from the control unit 45.

The drive unit 46 also includes an encoder 54 that detects the rotation angle of each joint 50. The encoder 54 outputs the detection result (the rotation angle of the joint 50) to the controller 45. The encoder 54 may be provided in each joint 50.

An operation unit for operating the pair of hand units 43 a and the manipulator arm 43 is connected to the control unit 45.
The operation unit 55 is, for example, a joystick, and an operator can manually operate the manipulator arm 43 remotely. However, the operation unit 55 is not limited to this case, and may have any configuration as long as the manipulator arm 43 can be operated.
The control unit 45 controls the hand actuator 52 and the joint actuator 53 based on the operation command from the operation unit 55, and controls the pair of hand units 43 a and the manipulator arm 43.

The optical microscope unit 44 includes an illumination system 60 arranged above the stage 42, an observation system 61 arranged below the stage 42 and observing the cells C from below the cell 41 through a through hole 42 a of the stage 42, and an observation system 61. And a display unit 62 for displaying an image observed by the system 61.

The illumination system 60 includes a light source (not shown) and various condenser lenses (not shown) for collecting light from the light source on the cells C. The observation system 61 includes an objective lens (not shown) and various imaging lenses for focusing the focal point of the objective lens on the cells C, and outputs the observed image to the display unit 62.
Thereby, the operator can operate the operation unit 55 while confirming the state of the cells C, the positional relationship between the cells C and the biological sample knife 1, and the like by the image displayed on the display unit 62.

The observation system 61 including the objective lens is not limited to being arranged below the stage 42, and may be arranged above the stage 42.

(Cleavage of cells)
Next, a case will be described in which the manipulator arm 43 configured as described above is operated to cut the cells C with the biological sample knife 1.
In the biological sample knife 1, the base end portion 3b of the holding member 3 is cantilevered by the manipulator arm 43 via the pair of hand portions 43a.

The operator observes the state of the cells C in the cells 41 with an optical microscope and identifies the position where the cells C are cut while confirming the observation image on the display unit 62. After the cutting position of the cell C is specified, the manipulator arm 43 is three-dimensionally operated via the operation unit 55 to bring the blade portion 12 of the biological sample knife 1 close to the cutting position. At this time, since the positional relationship between the cells C and the cutting edge portion 12 can be confirmed on the display unit 62, the cutting edge portion 12 can be accurately approached to the cutting position.

Next, while checking the image on the display unit 62, the manipulator arm 43 is operated via the operation unit 55 to cut the cells C by the biological sample knife 1. As a result, it is possible to achieve the same effect as that of the first embodiment.
Particularly, in the case of this embodiment, since the manipulator arm 43 is used, a finer and more accurate cutting operation can be performed. Moreover, since the holding member 3 is transparent, the knife portion 2 can be confirmed without being affected by the holding member 3. Therefore, the knife portion 2 can be accurately approached to the desired cutting position of the cell C.

The technical scope of the present invention is not limited to the above-described embodiment, and various modifications can be made without departing from the spirit of the present invention.

For example, in the above-described embodiment, cells have been described as an example of the biological sample, but the biological sample is not limited to cells. For example, it may be a living tissue obtained from a human body, an animal, a plant or the like.
Furthermore, the object that can be cut by the knife is not limited to a biological sample, and may be, for example, a metal film. That is, since the knife according to the present invention has high strength, high corrosion resistance, high heat resistance, and non-magnetism, it can damage the substrate even when cutting a metal pattern on the substrate, for example. It becomes possible to cut and process a metal pattern into a fine pattern. Further, according to the above-described manufacturing method, the knife can be easily manufactured by the FIB processing apparatus, so that the manufacturing cost can be suppressed. Further, the object is not limited to the metal film, and may be a resin or an elastic material.

In the above embodiment, the case where the knife portion is formed in the shape of a triangular pyramid has been described as an example, but the present invention is not limited to this case.
For example, as shown in FIG. 16, a knife formed such that the knife thickness T in the crest portion 13 is uniform over the entire length, and the knife thickness T gradually decreases from the crest portion 13 toward the cutting edge portion 12. A biological sample knife (knife) 70 including the portion 71 may be used.
Even in this case, the knife thickness T in the cutting edge portion 12 can be made extremely thin, for example, about 100 nm, and the knife portion 71 can be elastically deformed in the knife width W direction (vertical direction). The part 12 can be protected, and the stress applied to the cells C can be suppressed, and the cells C can be cut with good sharpness with little resistance.

Further, as shown in FIG. 17, a biological sample knife in which the reflecting surface 80 that reflects the detection light B emitted from the outside and faces different directions according to the elastic deformation of the knife portion 2 is formed in the ridge portion 13. You can set it to 1.

In this case, for example, the semiconductor laser light source 81 can emit laser light as the detection light B toward the reflecting surface 80, and the laser light reflected by the reflecting surface 80 can be received by the laser light receiving portion 82. The laser light receiving unit 82 is, for example, a four-division photo detector, and is capable of detecting the amount of change in elastic deformation of the knife unit 2 based on the incident position of the laser light.
Therefore, the elastically deformed state of the knife portion 2 can be grasped, and the cell C can be cut while grasping the state of the knife portion 2 more accurately.

Although the elastic deformation of the knife portion 2 is detected by the so-called optical lever method using the reflection surface 80 formed on the peak 13 and the detection light B as described above, the invention is not limited to the optical lever method. For example, the deflection of the knife portion 2 may be measured by a self-detection method in which the knife portion 2 itself is provided with a displacement detection mechanism (for example, a piezoresistive element).

Besides, it is possible to appropriately replace the constituent elements in the embodiment with known constituent elements within the scope of the present invention, and it is also possible to appropriately combine the modified examples described above.

L... Knife length T... Knife thickness W... Knife width θ1... First tilt angle θ2... Second tilt angle 1, 70... Knife for biological sample (knife)
2, 71... Knife part 3... Holding member 3a... Holding member tip part 3b... Holding member base end part 10... Knife base end part 11... Knife tip part 12... Blade tip part 13... Ridge part 14... Tip part 15... Knife body 80... Reflective surface

Claims (7)

Formed from a Co-Ni based alloy containing Co, Ni, Cr and Mo, and provided with a knife portion extending in one direction from a knife base end supported in a cantilever manner to a knife tip,
In the knife portion, a blade tip portion that comes into contact with an object, and a peak portion that is arranged to face the blade tip portion are formed over the entire length,
The knife thickness in the ridge is 30 μm or less,
The knife portion is formed such that the knife thickness gradually decreases from the crest portion toward the cutting edge portion. The knife according to claim 1,
A knife having a knife thickness of 100 nm or less at the cutting edge. The knife according to claim 1,
The knife length of the knife part is 50 μm or less,
The knife width at the knife base end is 20 μm or less,
The knife thickness in the ridge is 10 μm or less,
A knife having a knife thickness of 100 nm or less at the cutting edge. The knife according to any one of claims 1 to 3,
The knife part is
The knife is formed such that the knife thickness and the knife width from the peak portion to the cutting edge portion are gradually reduced from the knife base end portion toward the knife tip end portion. The knife according to claim 4,
The knife part is
A pointed portion that is located on the knife tip portion side and that the blade tip portion is inclined at a first inclination angle with respect to the crest portion;
A knife main body that is continuously provided to the tip portion in a state of being located between the tip portion and the knife base portion, and the blade tip portion is inclined at a second inclination angle with respect to the peak portion,
The knife, wherein the first tilt angle is greater than the second tilt angle. The knife according to any one of claims 1 to 5,
A knife having reflection surfaces that reflect detection light emitted from the outside and that face different directions depending on the deformation of the knife portion. The knife according to any one of claims 1 to 6,
A holding member that is formed to extend in the extending direction of the knife portion from the base end portion toward the tip end portion, and that the base end portion is held in a cantilevered manner,
The knife portion is attached to the holding member by attaching the knife base end portion to the tip end portion,
The knife, wherein the holding member is made of a transparent material.

Images