JP7285306B2 - Ultramicrotome for 3D imaging - Google Patents

Description

The present invention relates to a three-dimensional imaging ultramicrotome, and more particularly to a three-dimensional imaging ultramicrotome that thins a specimen to produce a three-dimensional image of the specimen.

The technical demand for constructing a three-dimensional image of a bio-specimen is increasing further, and in the technique of constructing a three-dimensional image using an electron microscope, the specimen is cut into ultra-thin sections, and the cutting surface is A three-dimensional image of the specimen is obtained by repeating the scanning operation. Therefore, there is a need for an ultramicrotome capable of thinning a test piece to the order of nanometers.

Ultramicrotome for three-dimensional imaging needs a drive system that can realize precise cutting motion of the blade to improve the quality of cut surface preparation, such as serial block face scanning electron microscope (SBF-SEM). The size is limited so that it can work with the three-dimensional imaging device as a stage for the three-dimensional imaging device.

The present invention is an ultramicrotome for thinning a test piece for three-dimensional imaging of the test piece, which has a compact structure so as to be inserted into a narrow space provided in a three-dimensional imaging apparatus and has low vibration. To provide an ultramicrotome for three-dimensional imaging in which cutting motion of is realized by one degree of freedom.

A three-dimensional imaging ultramicrotome according to one embodiment of the present invention is an ultramicrotome for thinning a test piece that is an observation target of a three-dimensional imaging apparatus, comprising: a test piece table to which the test piece is fixed; a test piece storage section having a movement adjusting means for adjusting the movement of the test piece table; a cutting section including a knife having a blade at one end for cutting the test piece; and a knife holder for fixing the knife; a power generation unit for generating driving energy so that the blade cuts the test piece; and a power transmission unit for transmitting the driving energy to the cutting unit, wherein the power generation unit and the power transmission The parts are connected by a wire by which the driving energy can be transmitted to the cutting part.

further comprising a base on which the test piece storage unit, the power generation unit, and the power transmission unit are placed; can be accommodated in space.

It may further include an observation unit including at least one camera for observing an approaching state of the blade to the test piece.

The observation unit includes a first camera that observes the test piece from a first direction, and a second camera that observes the test piece from a second direction having a certain angle with respect to the first direction. can be done.

The power transmission part has a support means to which the cutting part is fixed, a free end connected to the support means at one end and a fixed end at the other end, and the rotation of the free end has a single degree of freedom. and a rotational deformation of the free end of the leaf spring can effect a cutting motion of the blade.

A free end of the leaf spring is connected to one end of the wire, and the power generator winds the other end of the wire to cause rotational deformation of the free end.

The power generator may include a motor that provides rotational power, and a gear that transmits the rotational power of the motor to a shaft around which the wire is wound.

The test strip receiving part, the power generating part, and the power transmitting part may be arranged on a base, and at least one damper may be provided between the power generating part and the base.

The three-dimensional imaging ultramicrotome according to the present invention has a compact design for transmitting power to the blade that slices the test piece, so that it can be inserted into a storage space provided in a conventional three-dimensional imaging device, It can be easily linked with the device as a stage for the device.

In addition, in the three-dimensional imaging ultramicrotome according to the present invention, the blade operates with one degree of freedom, so that a plurality of cutting motions draws a specified trajectory, and power is transmitted to the blade. The configurations are designed to reduce vibration so that the specimen can be cut flat.

In addition, the three-dimensional imaging ultramicrotome according to the present invention has a configuration that allows observation of the approach between the blade and the test piece, so that after the microtome is accommodated in the three-dimensional imaging device, cutting of the test piece Work can be done immediately.

The effects obtained from the present invention are not limited to the effects mentioned above, and further effects not mentioned will be clearly understood by those having ordinary knowledge in the technical field to which the present invention belongs from the following description. .

1 is a perspective view of an ultramicrotome for three-dimensional imaging according to one embodiment of the present invention; FIG. 1 is a top view of an ultramicrotome for three-dimensional imaging according to one embodiment of the present invention; FIG. Fig. 2 shows a leaf spring of an ultramicrotome for three-dimensional imaging according to one embodiment of the invention; 1 is a perspective view showing how a three-dimensional imaging ultramicrotome according to one embodiment of the present invention is housed in a housing space provided in a three-dimensional imaging apparatus. FIG. 1 is a front view of how a three-dimensional imaging ultramicrotome according to an embodiment of the present invention is housed in a housing space provided in a three-dimensional imaging apparatus; FIG. Fig. 3 shows the state before a cutting motion of a three-dimensional imaging ultramicrotome according to an embodiment of the invention; Fig. 6b shows the specimen of Fig. 6a; Fig. 3 shows a three-dimensional imaging ultramicrotome during a cutting motion according to an embodiment of the present invention; Fig. 7b shows the specimen of Fig. 7a;

Preferred embodiments of the present invention will now be described in detail with reference to the accompanying drawings.

The terms used herein are terms defined in view of the functions of the present invention, which may vary according to the intentions or practices of users, operators. Therefore, definitions of such terms should be based on the overall content of this specification.

It should be noted that the embodiments disclosed below are not intended to limit the scope of rights of the present invention, but are merely exemplary matters of the components presented in the claims of the present invention. Embodiments that are included in the technical ideas throughout the specification and that include components that can be replaced as equivalents in the components of the claims may be included in the scope of the present invention.

And terms such as "first", "second", "one side", and "other side" in the embodiments disclosed below are used to distinguish one component from other components. and its components are not limited by the term. In the following description of the present invention, detailed descriptions of known techniques that may obscure the gist of the present invention will be omitted.

1 and 2 are a perspective view and a top view of an ultramicrotome for three-dimensional imaging according to one embodiment of the present invention.

Referring to FIGS. 1 and 2, the three-dimensional imaging ultramicrotome according to one embodiment of the present invention includes a test piece storage unit 100, a cutting unit 200, a power generation unit 300, a power transmission unit 400, a wire 700, for thinning the test piece to be observed by the three-dimensional imaging device to a thickness in nanometer units. At this time, the microtome cuts the specimen into ultra-thin slices, and the three-dimensional imaging device repeats the process of scanning the cut surfaces, thereby generating a three-dimensional image of the specimen. Here, the microtome is linked with the three-dimensional imaging apparatus as a stage of the three-dimensional imaging apparatus, and sequentially provides the cutting surface of the test piece to be three-dimensionally imaged.

In one embodiment, the test strip container 100 includes a test strip table 110 for fixing the test strip 10 and a movement adjusting means 120 for adjusting the movement of the test strip table 110 .

The movement adjusting means 120 adjusts the movement of the test piece stage so that the test piece moves in three dimensions, that is, in x, y, and z directions. In order for the microtome to form the first cut surface and the next cut surface on the specimen, the specimen is cut to a height in nanometers, i.e., the cut thickness required for three-dimensional imaging. should move in the z-axis direction by For this purpose, the movement adjusting means 120 is provided with an automated device that enables minute movement of the test piece stage. For example, the automation device can be a piezo-type actuator that is operated piezoelectrically.

In the illustrated embodiment, the movement adjusting means 120 includes movement adjusting screws 121, 122 for moving the test piece 10 and the test piece table 110 in the x-axis and y-axis directions. By manually operating the movement adjusting screws 121 and 122, the test piece table 110 is moved to the x position so that the test piece 10 is in contact with the blade 211, which will be described later, that is, so that the test piece is on the movement locus for cutting by the blade. Move in the axial and y-axis directions.

The cutting part 200 includes a knife 210 having a blade 211 for cutting a test piece at one end, and a knife holder 220 fixing the knife 210 . Blade 211 can be made of diamond. The cutting part 200 is detachably fixed to a support means 430 included in the power transmission part 400 to be described later, and moves in the air along with the movement of the support means 430 .

The power generation unit 300 generates driving energy such that the blade cuts the test piece. In one embodiment, the power generator 300 includes a motor 310 that provides rotational force and a gearbox 320 that transmits the rotational force of the motor. The gears in the gearbox 320 may be worm gears in which a worm wheel and a worm are meshed with each other to form a pair. The rotational force of the motor 310 is transmitted to a shaft around which a wire 700 (to be described later) is wound via a worm gear, thereby winding or unwinding the wire 700 .

The power transmission unit 400 transmits driving energy generated by the power generation unit 300 to the cutting unit 200 . The power transmission unit 400 includes support means 430 to which the cutting part 200 is fixed, leaf springs 410 and fixing means 420 .

The support means 430 is configured to be combined with the cutting part 200, and includes a plate on which the knife holder 220 is mounted and a coupling means such as a screw to attach/detach and fix the knife holder 220. As shown in FIG. The support means 430 is connected to a free end of a leaf spring 410 to be described later, and is constrained to follow the rotational movement due to the deformation of the leaf spring 410 . As a result, the trajectory of the cutting motion of the blade 211 of the cutting portion is restrained by the rotational movement caused by the deformation of the leaf spring.

The fixing means 420 is fixed to the base 500 described below and includes a fixing structure 440 having an appearance corresponding to the shape of the fixed end so as to fix the fixed end of the leaf spring 410 .

FIG. 3 shows a leaf spring of an ultramicrotome for three-dimensional imaging according to one embodiment of the invention.

Referring to FIG. 3, one end of the leaf spring 410 is a fixed end and the other end is a free end. A fixed end 411 of the leaf spring 410 has a fixed position coupled with the fixed structure 440 . A free end 412 of leaf spring 410 is connected to wire 700 and support means 430 . When the power generator 300 winds the wire 700 , the pulling force (F) of the wire 700 is applied to the free end 412 of the leaf spring, and the deflection of the leaf spring 410 is induced, causing the free end 412 to rotate.

The deformation of the leaf spring 410 according to one embodiment of the invention, namely the rotational deformation (O) of the free end 412, has a single degree of freedom. In other words, the leaf spring 410 is structurally only capable of deformation constrained to a single degree of freedom, that is, rotation of the free end, and absolutely eliminates degrees of freedom for deformation such as plate distortion. be done. Thereby, the cutting motion of the blade has a single degree of freedom corresponding to the rotational deformation of the free end of the leaf spring, and always draws the same movement trajectory in multiple cutting operations.

In addition, since the force (F) that induces the deformation of the leaf spring 410 is transmitted through the wire 700, the vibration generated in the power generation unit 300 causes the deformation of the leaf spring 410 and the rotation of the free end of the leaf spring. Impact on movement is minimized. As a result, the transmission of the driving energy through the wire minimizes the influence of the vibration generated by the power generator 300 on the cutting motion of the blade and ensures that the cutting motion of the blade is performed on a constant trajectory. . The wire 700 can be made of a material with very little elasticity.

1 and 2, the three-dimensional imaging microtome 1000 according to one embodiment of the present invention further includes a base 500 and an observation part 600. As shown in FIG.

The base 500 is a rigid plate on which the test strip receiving part 100, the power generating part 300, and the power transmitting part 400 are arranged. The base 500 includes connecting and fixing means so that some components included in the test strip receiving portion 100, the power generating portion 300, and the power transmitting portion 400 are directly and fixedly connected on the base 500. FIG. In the illustrated embodiment, the movement adjusting means 120 of the test strip receiving portion 100, the gears of the power generating portion 300 and the fixing means 420 of the power transmitting portion 400 are arranged on this base 500. FIG. Stands 611 and 621 to which clamps for supporting cameras included in an observation unit 600 to be described later are attached are arranged on the base 500 .

When the gearbox 320 containing the gears of the power generator 300 is placed on the base 500, the vibration absorbers are used to minimize the transmission of vibrations generated by the power generator to other components of the microtome 1000. At least one damper 330 for is provided between the gearbox 320 and the base 500 .

The base 500 is provided with connecting means for attaching the microtome 1000 to a three-dimensional imaging apparatus. As an example, the connecting means is a through-hole having an inner wall formed with a male screw and a female screw thread that engages with the screw thread of the male screw. can be.

The observation unit 600 is configured to observe the blade 211 approaching the test piece 10 and includes at least one camera. The observation unit 600 provides an image including the blade and the test piece, thereby assisting adjustment of the movement adjusting means 120 so that the test piece is placed on the moving trajectory of the blade. This provides the advantage that the operation of cutting the specimen can be done immediately after the microtome is housed in the .

As an example, the observation unit 600 includes a first camera 610 viewing the test piece from a first direction and a second camera 620 viewing the test piece from a second direction having a certain angle with respect to the first direction. A first camera 610 and a second camera 620 are supported by clamps attached to stands 611, 612 located on the base 500 and provide images of the periphery of the specimen in first and second directions, respectively. At this time, the test piece is a very small object that cannot be visually observed, and the distance difference between the test piece and the blade is in the order of nanometers or micrometers, so the test piece and the test piece and the blade First camera 610 and second camera 620 have a minimum working distance of 33.5 mm, a maximum magnification of 30×, and a viewing angle of 13 mm×9.7 mm ( Field of View (FOV) requirements.

4 and 5 are a perspective view and a front view, respectively, showing how a three-dimensional imaging ultramicrotome according to one embodiment of the present invention is housed in a housing space provided in a three-dimensional imaging apparatus.

Referring to FIGS. 4 and 5, the three-dimensional imaging ultramicrotome 1000 according to the present invention continuously provides a three-dimensional imaging device with a sample to be observed and a cut surface of the sample. The stage is housed in a housing space provided in the three-dimensional imaging apparatus.

As an example, the three-dimensional imaging device is a scanning electron microscope, and the microtome 1000 is housed in a vacuum chamber of the scanning electron microscope. The microtome 1000 is connected to the vacuum chamber lid 22 by connecting means provided on the base 500 and enters the vacuum chamber body 21 .

The size of the microtome 1000 is limited because the space provided in the three-dimensional imaging apparatus for accommodating the microtome is limited. Therefore, the ultramicrotome for three-dimensional imaging according to the present invention is designed with a plurality of configurations so as to have a compact structure due to the limitation of the accommodation space.

Figure 6a shows the state before the cutting motion of a three-dimensional imaging ultramicrotome according to an embodiment of the invention, and Figure 6b shows the specimen of Figure 6a. FIG. 7a shows a three-dimensional imaging ultramicrotome according to an embodiment of the invention during a cutting motion, and FIG. 7b shows the specimen of FIG. 7a.

6a to 7b, a three-dimensional imaging ultramicrotome 1000 according to one embodiment of the present invention is placed in the housing space of a three-dimensional imaging apparatus and repeats the operating states of FIGS. 6a and 7a. As shown in FIGS. 6a and 6b, the coiling of the wire 700 induces deformation of the leaf spring 410 with the blade 211 and the test piece 10 at a constant distance. As shown in Figures 7a and 7b, the blade 211 of the cutting part 200 connected to the free end 412 of the leaf spring 410 moves along the path of movement of the free end 412 having a single degree of freedom. The test piece 10 is cut while drawing, and the cut surface is provided to a three-dimensional imaging device. A scanning unit 23 of the three-dimensional imaging device scans the cutting surface of the specimen provided by the blade.

1000 Ultramicrotome for three-dimensional imaging 100 Specimen housing section 200 Cutting section 300 Power generation section 400 Power transmission section 500 Base 600 Observation section 700 Wire 10 Specimen

Claims (8)

An ultramicrotome for thinning a test piece to be observed by a three-dimensional imaging device,
a test strip holder having a test strip table to which the test strip is fixed and a movement adjusting means for adjusting the movement of the test strip table;
a cutting unit including a knife having a blade at one end for cutting the test piece, and a knife holder for fixing the knife;
a power generator for generating drive energy such that the blade is in motion to cut the specimen;
two leaf springs formed to cross each other, each including a fixed end coupled to a fixed structure and a free end coupled to a support means formed in the cutting portion;
a power transmission unit that transmits the driving energy to the cutting unit;
The power generation unit and the power transmission unit are connected by a wire, and one end of the wire is connected to a free end of the leaf spring to cause rotational deformation of the free end, thereby transferring the driving energy to the cutting unit. A transmitted, ultramicrotome for three-dimensional imaging. further comprising a base on which the test strip housing portion, the power generation portion, and the power transmission portion are placed;
2. The ultramicrotome for three-dimensional imaging according to claim 1, wherein said microtome is housed in a housing space provided in said three-dimensional imaging apparatus by connecting means provided on said base. 3. The ultramicrotome for three-dimensional imaging according to claim 1, further comprising an observation unit including at least one camera for observing the approach of said blade to said specimen. The observation unit is
a first camera that observes the test piece from a first direction;
4. The three-dimensional imaging ultramicrotome of claim 3, comprising a second camera that views the specimen from a second direction having an angle with respect to the first direction. The ultramicrotome for three-dimensional imaging according to claim 1, wherein the rotation of the free end of the leaf spring has a single degree of freedom, and the rotational deformation of the free end of the leaf spring constrains the cutting motion of the blade. . 2. The ultramicrotome for three-dimensional imaging according to claim 1, wherein said power generator winds the other end of said wire to cause rotational deformation of said free end. 7. The ultramicrotome for three-dimensional imaging according to claim 6, wherein the power generator includes a motor that provides rotational power, and a gear that transmits the rotational power of the motor to a shaft around which the wire is wound. further comprising a base on which the test strip housing portion, the power generation portion, and the power transmission portion are arranged;
8. The ultramicrotome for three-dimensional imaging according to claim 7, wherein at least one or more dampers are provided between the power generator and the base.

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