WO2022139235A1 - Ultramicrotome pour imagerie tridimensionnelle - Google Patents

Ultramicrotome pour imagerie tridimensionnelle Download PDF

Info

Publication number
WO2022139235A1
WO2022139235A1 PCT/KR2021/018099 KR2021018099W WO2022139235A1 WO 2022139235 A1 WO2022139235 A1 WO 2022139235A1 KR 2021018099 W KR2021018099 W KR 2021018099W WO 2022139235 A1 WO2022139235 A1 WO 2022139235A1
Authority
WO
WIPO (PCT)
Prior art keywords
specimen
unit
cutting
dimensional imaging
microtome
Prior art date
Application number
PCT/KR2021/018099
Other languages
English (en)
Korean (ko)
Inventor
허환
권희석
Original Assignee
한국기초과학지원연구원
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 한국기초과학지원연구원 filed Critical 한국기초과학지원연구원
Publication of WO2022139235A1 publication Critical patent/WO2022139235A1/fr

Links

Images

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/02—Devices for withdrawing samples
    • G01N1/04—Devices for withdrawing samples in the solid state, e.g. by cutting
    • G01N1/06—Devices for withdrawing samples in the solid state, e.g. by cutting providing a thin slice, e.g. microtome
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/02—Devices for withdrawing samples
    • G01N1/04—Devices for withdrawing samples in the solid state, e.g. by cutting
    • G01N1/06—Devices for withdrawing samples in the solid state, e.g. by cutting providing a thin slice, e.g. microtome
    • G01N2001/061—Blade details
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/02—Devices for withdrawing samples
    • G01N1/04—Devices for withdrawing samples in the solid state, e.g. by cutting
    • G01N1/06—Devices for withdrawing samples in the solid state, e.g. by cutting providing a thin slice, e.g. microtome
    • G01N2001/065—Drive details

Definitions

  • the present invention relates to an ultra microtome for three-dimensional imaging, and more particularly, to an ultra microtome for three-dimensional imaging that slices a specimen in order to generate a three-dimensional image of the specimen.
  • the technical demand for constructing a three-dimensional image of a bio-specimen is increasing, and the technology for constructing a three-dimensional image using an electron microscope cuts the specimen into an ultra-thin section and repeats the operation of scanning the cutting surface to achieve three-dimensional imaging of the specimen. get Therefore, an ultra-microtome capable of thinning specimens in nanometers is required.
  • Ultra microtome for 3D imaging requires a drivetrain that can implement precise cutting motion of the blade in order to improve the quality of the cutting surface, and the stage of 3D imaging devices such as continuous block surface scanning electron microscope (SBF-SEM).
  • SBF-SEM continuous block surface scanning electron microscope
  • the present invention is an ultra microtome for thinning a specimen for three-dimensional imaging of a specimen. It has a compact structure so that it can be inserted into a narrow space provided in a three-dimensional imaging device, and at the same time realizes a low-vibration cutting motion with one degree of freedom. Ultra microtome for 3D imaging is provided.
  • An ultra microtome is an ultra microtome for thinning a specimen to be observed by a three-dimensional imaging apparatus, and includes a specimen stand to which the specimen is fixed and a movement control means for controlling the movement of the specimen stand. wealth; a cutting unit including a knife having a blade for cutting the specimen at one end and a knife holder for fixing the knife; a power generating unit generating driving energy so that the blade performs a motion of cutting the specimen; and a power transmitting unit transmitting the driving energy to the cutting unit, wherein the power generating unit and the power transmitting unit are connected by a wire, and the driving energy may be transmitted to the cutting unit by the wire.
  • It may further include an observation unit including at least one camera for observing the state of approach of the blade to the specimen.
  • the observation unit may include a first camera for observing the specimen in a first direction and a second camera for observing the specimen in a second direction having a constant angle with respect to the first direction.
  • the power transmission unit includes a supporting unit to which the cutting unit is fixed, and a leaf spring having one end fixed end and a free end connected to the supporting unit at the other end, and rotation of the free end has a single degree of freedom, the plate A cutting motion of the blade may be achieved by rotational deformation of the free end of the spring.
  • the free end of the leaf spring may be connected to one end of the wire, and the power generating unit may wind the other end of the wire to cause rotational deformation of the free end.
  • the power generating unit may include a motor providing rotational power and a gear transmitting rotational power of the motor to a shaft around which the wire is wound.
  • It may further include a base in which the specimen receiving unit, the power generating unit, and the power transmitting unit are disposed, and at least one damper may be provided between the power generating unit and the base.
  • the ultra microtome for 3D imaging according to the present invention is inserted into the receiving space provided in the existing 3D imaging device by designing compactly the components that transmit power to the blade for flaking the specimen, and the device and the device as an offering of the device can be easily linked.
  • the ultra microtome for three-dimensional imaging draws a designated work trajectory in several cutting motions as the blade operates with one degree of freedom, and the components that transmit power to the blade are designed to reduce vibration, thereby cutting the specimen. It can be cut flat.
  • the ultra microtome for 3D imaging has a configuration that can observe the approach between the blade and the specimen, so that the cutting operation of the specimen can be performed immediately after the microtome is accommodated in the 3D imaging device.
  • FIG. 1 is a perspective view of an ultra microtome for 3D imaging according to an embodiment of the present invention.
  • FIG. 2 is a top view of an ultra microtome for 3D imaging according to an embodiment of the present invention.
  • FIG 3 illustrates a leaf spring of an ultra microtome for three-dimensional imaging according to an embodiment of the present invention.
  • FIG 4 is a perspective view of a scene in which the ultra microtome for 3D imaging is accommodated in an accommodation space provided in the 3D imaging apparatus according to an embodiment of the present invention.
  • FIG 5 is a front view of a scene in which an ultra microtome for 3D imaging is accommodated in an accommodation space provided in a 3D imaging apparatus according to an embodiment of the present invention.
  • Figure 6a shows a state before the cutting motion of the ultra microtome for three-dimensional imaging according to an embodiment of the present invention.
  • Fig. 6b shows the specimen of Fig. 6a.
  • FIG. 7A shows a state during a cutting motion of an ultra microtome for three-dimensional imaging according to an embodiment of the present invention.
  • Fig. 7b shows the specimen of Fig. 7a.
  • 1 and 2 are a perspective view and a top view of an ultra microtome for 3D imaging according to an embodiment of the present invention.
  • the ultra microtome for 3D imaging includes a specimen receiving unit 100 , a cutting unit 200 , a power generating unit 300 , and a power transmitting unit 400 . , and a wire 700, and the specimen, which is an observation object of the three-dimensional imaging apparatus, is sliced to a thickness of nanometers.
  • the microtome cuts the specimen into an ultra-thin section, and the three-dimensional imaging device repeats the process of scanning the cutting surface to generate a three-dimensional image of the specimen.
  • the microtome is linked to the three-dimensional imaging apparatus as a product of the three-dimensional imaging apparatus and sequentially provides a cutting surface of a specimen as a three-dimensional imaging object.
  • the specimen receiving unit 100 includes a specimen stand 110 for fixing the specimen 10 and a movement adjusting means 120 for controlling the movement of the specimen stand 110 .
  • the movement control means 120 controls the movement of the specimen stand so that the specimen moves in three dimensions, that is, in the x, y, and z axis directions.
  • the microtome forms a first cutting surface on the specimen, and in order to form the next cutting surface, the specimen must be moved in the z-axis direction by a height of nanometers, that is, a cutting thickness required for three-dimensional imaging.
  • the movement control means 120 is provided with an automation device that allows the specimen table to be moved minutely.
  • the automation device may be a piezo-type actuator operated in a piezoelectric manner.
  • the movement adjustment means 120 includes movement adjustment screws 121 and 122 for moving the specimen 10 and the specimen stand 110 in the x-axis and y-axis directions.
  • the specimen 10 is in contact with the blade 211 to be described later, that is, the specimen stand 110 through the manual operation of the movement adjusting screws 121 and 122 so that the specimen can exist on the movement trajectory for cutting the blade. ) moves in the x-axis and y-axis directions.
  • the cutting unit 200 includes a knife 210 having a blade 211 for cutting a specimen at one end and a knife holder 220 for fixing the knife 210 .
  • the blade 211 may be made of a diamond material.
  • the cutting part 200 is detachably fixed to the 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 generating unit 300 generates driving energy so that the blade cuts the specimen.
  • the power generating unit 300 includes a motor 310 for providing a rotational force and a gear box 320 for transmitting the rotational force of the motor.
  • the gear in the gear box 320 may be a worm gear in which a worm wheel and a worm are meshed with each other.
  • the rotational force of the motor 310 is transmitted to a shaft on which a wire 700 to be described later is wound through a worm gear, and eventually winds or unwinds the wire 700 .
  • the power transmitting unit 400 transmits the driving energy generated by the power generating unit 300 to the cutting unit 200 .
  • the power transmission unit 400 includes a support unit 430 to which the cutting unit 200 is fixed, a leaf spring 410 , and a fixing unit 420 .
  • the support means 430 is configured to be coupled to the cutting part 200 , and includes coupling means, for example, screws, etc. to detach and fix the plate on which the knife holder 220 is seated and the knife holder 220 .
  • the support means 430 is connected to the free end of the leaf spring 410 to be described later, and is constrained to and follows rotational movement due to the deformation of the leaf spring 420 . Accordingly, the cutting motion trajectory of the blade 211 of the cutting part is constrained to rotational movement due to the deformation of the leaf spring.
  • the fixing means 420 is fixed to the base 500 to be described later, and includes a fixing structure 440 having an appearance corresponding to the shape of the fixing end to fix the fixed end of the leaf spring 410 .
  • FIG 3 illustrates a leaf spring of an ultra microtome for three-dimensional imaging according to an embodiment of the present invention.
  • the leaf spring 410 is formed at one end as a fixed end and the other end as a free end.
  • the fixed end 411 of the leaf spring 410 is coupled to the fixing structure 440 and has a fixed position.
  • the free end 412 of the leaf spring 410 is connected to the wire 700 and the supporting means 430 .
  • the wire 700 applies a pulling force F to the free end 412 of the leaf spring, and the bending of the leaf spring 410 is induced and the free end ( 412) rotate.
  • the deformation of the leaf spring 410 according to an embodiment of the present invention that is, the rotational deformation O of the free end 412 has a single degree of freedom.
  • the leaf spring 410 is structurally limited to a single degree of freedom, that is, only rotation of the free end is possible, and the degree of freedom against deformation such as distortion of the plate is absolutely excluded. Accordingly, 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 several cutting operations.
  • the vibration generated in the power generating unit 300 causes the deformation of the leaf spring 410 and the free end of the leaf spring.
  • the effect on the rotational motion of As a result, the driving energy transfer through the wire minimizes the influence of the vibration generated by the power generating unit 300 on the cutting motion of the blade and ensures that the blade cutting motion is made on a constant trajectory.
  • the wire 700 may be made of a material with very little elasticity.
  • the microtome 1000 for 3D imaging further includes a base 500 and an observation unit 600 .
  • the base 500 is a plate made of a hard material on which the specimen receiving unit 100 , the power generating unit 300 , and the power transmitting unit 400 are disposed.
  • the base 500 includes a connection and fixing means so that some components included in the specimen receiving unit 100 , the power generating unit 300 , and the power transmitting unit 400 are directly fixedly connected to the base 500 .
  • the movement control unit 120 of the specimen receiving unit 100, the gear of the power generating unit 300, and the fixing unit 420 of the power transmitting unit 400 are disposed on the base 500 do.
  • stands 611 and 621 equipped with clamps for supporting a camera included in an observation unit 600 to be described later are disposed on the base 500 .
  • the base 500 includes a connection means for the microtome 1000 to be mounted on the three-dimensional imaging apparatus, and in one example, the connection means may be a male screw and a through hole formed in an inner wall of a female screw thread engaged with the male screw thread. .
  • the observation unit 600 is configured to observe the state of approach of the blade 211 to the specimen 10 and includes at least one camera.
  • the observation unit 600 helps to adjust the movement control means 120 so that the specimen is placed on the movement trajectory of the blade by providing an image containing the blade and the specimen, and after the microtome is accommodated in the three-dimensional imaging device, the specimen It offers the advantage of being able to perform cutting operations immediately.
  • the observation unit 600 includes a first camera 610 that looks at the specimen in a first direction and a second camera 620 that looks at the specimen in a second direction having a constant angle with respect to the first direction.
  • the first camera 610 and the second camera 620 are supported by clamps mounted on stands 611 and 612 disposed on the base 500, and images around the specimen in the first and second directions, respectively. provides At this time, the specimen is a very small object that cannot be observed with the naked eye, and the difference between the distance between the specimen and the blade is also on the nanometer or micrometer level.
  • the first camera 610 and the second camera 620 have a minimum working distance of 33.5 mm, a maximum magnification of 30 times, and a field of view (FOV) of 13mm x 9.7mm.
  • FOV field of view
  • FIGS. 4 and 5 are perspective and front views, respectively, of a scene in which an ultra microtome for 3D imaging is accommodated in an accommodation space provided in a 3D imaging apparatus according to an embodiment of the present invention.
  • the ultra microtome 1000 for 3D imaging according to the present invention is provided as an object to the 3D imaging device to continuously provide a sample to be observed and a cutting surface of the sample to the 3D imaging device. It is accommodated in the receiving space provided in the dimensional imaging device.
  • the three-dimensional imaging apparatus is a scanning electron microscope
  • the microtome 1000 is accommodated into a vacuum chamber of the scanning electron microscope.
  • the microtome 1000 is coupled to the cover 22 of the vacuum chamber by a connecting means provided on the base 500 and enters the body 21 of the vacuum chamber.
  • the size of the microtome 1000 is limited. Therefore, in the ultra microtome for 3D imaging according to the present invention, a plurality of components are each designed to have a compact structure according to the limitation of the accommodation space.
  • FIG. 6A shows a state before the cutting motion of the ultra microtome for 3D imaging according to an embodiment of the present invention
  • FIG. 6B shows the specimen of FIG. 6A
  • 7A shows a state during a cutting motion of the ultra microtome for 3D imaging according to an embodiment of the present invention
  • FIG. 7B shows the specimen of FIG. 7A.
  • the ultra microtome 1000 for 3D imaging is disposed in the receiving space of the 3D imaging apparatus and repeats the operation states of FIGS. 6A and 7A .
  • the deformation of the leaf spring 410 is induced by the winding of the wire 700 in a state in which the blade 211 is spaced apart from the specimen 10 at a certain distance.
  • the blade 211 of the cutting part 200 connected to the free end 412 of the leaf spring 410 follows the movement path of the free end 410 having a single degree of freedom.
  • the specimen 100 is cut while drawing a movement trajectory, and a cutting surface is provided to the 3D imaging apparatus.
  • the scanning unit 23 of the 3D imaging apparatus scans the cutting surface of the specimen provided by the blade.

Landscapes

  • Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Sampling And Sample Adjustment (AREA)

Abstract

L'invention concerne un ultramicrotome pour imagerie tridimensionnelle, servant à trancher en sections ultra-minces un échantillon à observer par un dispositif d'imagerie tridimensionnelle, et comprenant : une partie de réception d'échantillon comprenant un étage d'échantillon sur lequel est fixé un échantillon et un moyen de commande de mouvement servant à commander le mouvement de l'étage d'échantillon ; une partie de coupe comprenant un dispositif de coupe pourvu d'une lame pour couper l'échantillon à une extrémité de celui-ci, et un support de dispositif de coupe destiné à immobiliser le dispositif de coupe ; une partie de génération d'énergie destinée à générer une énergie d'entraînement de sorte à permettre à la lame d'effectuer des mouvements pour couper l'échantillon ; et une partie de transmission de puissance destinée à transmettre l'énergie d'entraînement à la partie de coupe, la partie de génération d'énergie et la partie de transmission de puissance étant reliées par un fil, et l'énergie d'entraînement étant transmise à la partie de coupe par l'intermédiaire du fil.
PCT/KR2021/018099 2020-12-22 2021-12-02 Ultramicrotome pour imagerie tridimensionnelle WO2022139235A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR10-2020-0180739 2020-12-22
KR1020200180739A KR102254822B1 (ko) 2020-12-22 2020-12-22 3차원 이미징용 울트라 마이크로톰

Publications (1)

Publication Number Publication Date
WO2022139235A1 true WO2022139235A1 (fr) 2022-06-30

Family

ID=76152556

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2021/018099 WO2022139235A1 (fr) 2020-12-22 2021-12-02 Ultramicrotome pour imagerie tridimensionnelle

Country Status (3)

Country Link
JP (1) JP7285306B2 (fr)
KR (1) KR102254822B1 (fr)
WO (1) WO2022139235A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116448795A (zh) * 2022-12-30 2023-07-18 浙江大学 可集成于扫描电镜中的冷冻超薄切片装置

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR102254822B1 (ko) * 2020-12-22 2021-05-24 한국기초과학지원연구원 3차원 이미징용 울트라 마이크로톰

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP4336577B2 (ja) * 2003-12-25 2009-09-30 フェザー安全剃刀株式会社 ミクロトーム
JP2015038479A (ja) * 2013-08-19 2015-02-26 ライカ マイクロシステムズ リミテッド シャンハイＬｅｉｃａ Ｍｉｃｒｏｓｙｓｔｅｍｓ Ｌｔｄ． Ｓｈａｎｇｈａｉ 質量バランス構造およびそれを用いるミクロトーム
US20170067800A1 (en) * 2014-05-12 2017-03-09 The United States Of America, As Represented By The Secretary, Department Of Health And Human Serv Miniature serial sectioning microtome for block-face imaging
KR101967681B1 (ko) * 2018-11-01 2019-08-13 한국기초과학지원연구원 울트라 마이크로톰 장치
JP2020518349A (ja) * 2017-04-27 2020-06-25 キュラデル、エルエルシー 光学イメージングにおいての測距
KR102254822B1 (ko) * 2020-12-22 2021-05-24 한국기초과학지원연구원 3차원 이미징용 울트라 마이크로톰

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS4715595Y1 (fr) * 1967-05-20 1972-06-01
SE316026B (fr) * 1968-07-02 1969-10-13 Lkb Produkter Ab
SE424023B (sv) * 1980-10-27 1982-06-21 Lkb Produkter Ab Anordning vid en mikrotom
US4377958A (en) * 1981-04-02 1983-03-29 The United States Of America As Represented By The Secretary Of The Department Of Health And Human Services Remotely operated microtome
SE500941C2 (sv) * 1989-08-16 1994-10-03 Algy Persson Förfarande och apparat för snittning av ett preparat

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP4336577B2 (ja) * 2003-12-25 2009-09-30 フェザー安全剃刀株式会社 ミクロトーム
JP2015038479A (ja) * 2013-08-19 2015-02-26 ライカ マイクロシステムズ リミテッド シャンハイＬｅｉｃａ Ｍｉｃｒｏｓｙｓｔｅｍｓ Ｌｔｄ． Ｓｈａｎｇｈａｉ 質量バランス構造およびそれを用いるミクロトーム
US20170067800A1 (en) * 2014-05-12 2017-03-09 The United States Of America, As Represented By The Secretary, Department Of Health And Human Serv Miniature serial sectioning microtome for block-face imaging
JP2020518349A (ja) * 2017-04-27 2020-06-25 キュラデル、エルエルシー 光学イメージングにおいての測距
KR101967681B1 (ko) * 2018-11-01 2019-08-13 한국기초과학지원연구원 울트라 마이크로톰 장치
KR102254822B1 (ko) * 2020-12-22 2021-05-24 한국기초과학지원연구원 3차원 이미징용 울트라 마이크로톰

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116448795A (zh) * 2022-12-30 2023-07-18 浙江大学 可集成于扫描电镜中的冷冻超薄切片装置

Also Published As

Publication number Publication date
JP2022099272A (ja) 2022-07-04
KR102254822B1 (ko) 2021-05-24
JP7285306B2 (ja) 2023-06-01

Similar Documents

Publication Publication Date Title
WO2022139235A1 (fr) Ultramicrotome pour imagerie tridimensionnelle
EP0864181B1 (fr) Etage plat de balayage destine a la microscopie a sonde balayee
EP1887403A1 (fr) Stéréomicroscope
US8822951B2 (en) Manipulator carrier for electron microscopes
EP1319968A2 (fr) Objectif pour microscope avec des lentilles déplaçables à moteur, microscope et méthode d'imagerie d'un échantillon
US6072622A (en) Operation microscope
WO2016052776A1 (fr) Dispositif de support triaxial de microscope électronique à structure simple
TW554177B (en) Microscope unit
JP2017044819A (ja) ステージ装置及び顕微鏡システム
EP0310514B2 (fr) Microscope binoculaire
JP2006218105A (ja) 立体観察装置
CN220473797U (zh) 一种光纤内窥镜探头光纤镜头对准粘接装置
CN116898541B (zh) 柔性丝状电极植入装置
CN109528148B (zh) 单摆杆腔镜夹持装置
US6984947B2 (en) Apparatus and method for adjusting components of an optical or mechanical system
GB2149506A (en) Ultrasonic microscope
EP0039775B1 (fr) Indicateur électromécanique
EP0445670B1 (fr) Dispositif d'extraction de pièces étrangères pour assemblage combustible pour la production d'énergie nucléaire
US3879107A (en) Microstereoscope
JP2003140053A (ja) 光学顕微鏡別軸一体型走査型プローブ顕微鏡
JP2009204657A (ja) 細胞把持・回転観察装置
CN217007843U (zh) 深度成像系统
US4266128A (en) Electron microscope with optical microscope
EP1843188B1 (fr) Stabilisateur pour examen in vivo
WO2020242194A1 (fr) Platine de microscope électromotrice de type deltabot

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 21911302

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 21911302

Country of ref document: EP

Kind code of ref document: A1