CN116448795A - Frozen ultrathin section device capable of being integrated in scanning electron microscope - Google Patents
Frozen ultrathin section device capable of being integrated in scanning electron microscope Download PDFInfo
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- CN116448795A CN116448795A CN202211727177.2A CN202211727177A CN116448795A CN 116448795 A CN116448795 A CN 116448795A CN 202211727177 A CN202211727177 A CN 202211727177A CN 116448795 A CN116448795 A CN 116448795A
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- 230000007246 mechanism Effects 0.000 claims abstract description 116
- 238000005520 cutting process Methods 0.000 claims abstract description 105
- 238000001816 cooling Methods 0.000 claims description 59
- 238000009413 insulation Methods 0.000 claims description 26
- 230000033001 locomotion Effects 0.000 claims description 11
- 239000003507 refrigerant Substances 0.000 claims description 9
- 230000001105 regulatory effect Effects 0.000 claims description 7
- 238000012544 monitoring process Methods 0.000 claims description 6
- 230000000903 blocking effect Effects 0.000 claims description 5
- 230000005540 biological transmission Effects 0.000 claims description 3
- 239000000523 sample Substances 0.000 abstract description 108
- 239000012472 biological sample Substances 0.000 abstract description 8
- 238000003384 imaging method Methods 0.000 abstract description 8
- 239000012520 frozen sample Substances 0.000 abstract description 7
- 238000000034 method Methods 0.000 abstract description 3
- 230000008014 freezing Effects 0.000 abstract 1
- 238000007710 freezing Methods 0.000 abstract 1
- 238000012512 characterization method Methods 0.000 description 5
- 230000033228 biological regulation Effects 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 238000012546 transfer Methods 0.000 description 3
- 230000009471 action Effects 0.000 description 2
- 230000001276 controlling effect Effects 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 239000002826 coolant Substances 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 238000005057 refrigeration Methods 0.000 description 2
- 239000004696 Poly ether ether ketone Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000006835 compression Effects 0.000 description 1
- 238000007906 compression Methods 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 230000017525 heat dissipation Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 229920002530 polyetherether ketone Polymers 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 238000004626 scanning electron microscopy Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N23/00—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00
- G01N23/22—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00 by measuring secondary emission from the material
- G01N23/2202—Preparing specimens therefor
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- 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
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/42—Low-temperature sample treatment, e.g. cryofixation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N23/00—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00
- G01N23/22—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00 by measuring secondary emission from the material
- G01N23/225—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00 by measuring secondary emission from the material using electron or ion
- G01N23/2251—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00 by measuring secondary emission from the material using electron or ion using incident electron beams, e.g. scanning electron microscopy [SEM]
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
- G01N2001/2873—Cutting or cleaving
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E30/00—Energy generation of nuclear origin
- Y02E30/10—Nuclear fusion reactors
Abstract
The application relates to the technical field of scanning electron microscopes, in particular to a frozen ultrathin section device which can be integrated in a scanning electron microscope. The frozen ultrathin section device includes: a base; the sample feeding mechanism is arranged on the base and comprises a sample table and a pushing assembly, and the pushing assembly is connected with the sample table and is used for pushing the sample table to move and feed samples; a cutting mechanism for cutting the sample; the driving mechanism is arranged on the base and can move relative to the base, and is connected with the cutting mechanism and used for driving the cutting mechanism to move and cut a sample; and the temperature adjusting mechanism is respectively in heat conduction connection with the sample table and the cutting mechanism and is used for adjusting the temperature of the sample and the temperature of the cutting mechanism. According to the method, sample feeding and continuous slicing can be achieved in a freezing environment, an effective structural support is provided for three-dimensional structural representation of a large biological sample, and the structure of the sample is prevented from being damaged, so that a scanning electron microscope can implement a sequence section imaging technology on the frozen sample.
Description
Technical Field
The application relates to the technical field of scanning electron microscopes, in particular to a frozen ultrathin section device which can be integrated in a scanning electron microscope.
Background
Based on the sequential section imaging technology of the scanning electron microscope, the three-dimensional reconstruction of the biological sample at the cell or even the tissue level can be realized in the scanning electron microscope in a mode of observing while slicing. However, the current sequential section imaging technology based on scanning electron microscope still can only process the resin embedded sample after drying and dehydration. Numerous studies have shown that: the biological sample can be changed in structure in the process of drying the biological sample, so that the real structure of the sample can not be observed finally, and great trouble is brought to related researchers.
After the 2017 Nobel prize is obtained from the refrigeration electron microscope technology, the refrigeration technology is rapidly applied to the field of biological sample characterization. However, there is still a lack of equipment for sequential sectional imaging of frozen samples in scanning electron microscopy, thereby also impeding further development of related research efforts.
Disclosure of Invention
Based on this, it is necessary to provide a frozen ultra-thin section device which can be integrated in a scanning electron microscope for slicing frozen samples to form serial sections without damaging the true structure of the samples.
Aiming at the technical problems, the application provides the following technical scheme:
a frozen microtome device integrable in a scanning electron microscope, the frozen microtome device comprising: a base; the sample feeding mechanism is arranged on the base and comprises a sample table and a pushing assembly, the sample table is used for supporting a sample to be scanned, and the pushing assembly is connected with the sample table and used for pushing the sample table to move and feed the sample; a cutting mechanism for cutting the sample so that a cross section of the sample appears; the driving mechanism is arranged on the base and can move relative to the base, and is connected with the cutting mechanism and used for driving the cutting mechanism to move and cut a sample; and the temperature adjusting mechanism is respectively in heat conduction connection with the sample table and the cutting mechanism and is used for adjusting the temperature of the sample and the temperature of the cutting mechanism.
In one embodiment, the cutting mechanism comprises: a knife rest connected with the driving mechanism and capable of moving relative to the sample stage; the cutting knife is fixedly connected with the knife rest and can cut a sample on the sample table when moving along with the knife rest, wherein the cutting knife is in heat conduction connection with the temperature adjusting mechanism.
In one embodiment, the tool holder comprises: a connecting frame connected to the driving mechanism and movable relative to the sample stage; and the heat insulation frame is respectively connected with the connecting frame and the cutting knife and is used for blocking heat conduction between the connecting frame and the cutting knife.
In one embodiment, the cutting mechanism further comprises an insulation block disposed between the cutting blade and the insulation holder for blocking heat transfer between the insulation holder and the cutting blade.
In one embodiment, the cutting blade includes: the knife handle is fixedly connected with the knife rest and is in heat conduction connection with the temperature adjusting mechanism; the tool bit is fixed on the tool shank, and the tool bit can cut the sample on the sample platform when moving along with the tool rest.
In one embodiment, the drive mechanism includes: the sliding block is arranged on the base and can move relative to the base, and the cutting mechanism is arranged on the sliding block and is connected with the sliding block; the screw rod is in transmission connection with the sliding block and is used for driving the sliding block to slide relative to the base; and the first driving piece is used for driving the screw rod to move.
In one embodiment, the temperature adjustment mechanism includes: the cooling base is in heat conduction connection with the cutting mechanism and is used for adjusting the temperature of the cutting mechanism; the cooling device is used for providing a refrigerant and refrigerating; and the cooling pipeline is at least partially arranged in the cooling base and the sample table, and the refrigerant conveyed by the cooling equipment circulates in the cooling pipeline to adjust the temperature of the cooling base and the sample table.
In one embodiment, the temperature adjustment mechanism further comprises a flexible heat conducting strip connected to the cooling base and the cutting mechanism for conducting heat to cool the cutting mechanism.
In one embodiment, the temperature adjustment mechanism further comprises: the first temperature monitor is used for monitoring the temperature of the cutting mechanism in real time; the second temperature monitor is used for monitoring the temperature of the sample table in real time; and the control system is respectively connected with the first temperature monitor, the second temperature monitor and the cooling equipment electrically and/or in a signal manner, and the control system controls and regulates the quantity of the refrigerant which is conveyed into the cooling pipeline by the cooling equipment in real time according to the temperature data of the first temperature monitor and the second temperature monitor.
In one embodiment, the temperature adjustment mechanism further comprises a thermal insulation base mounted on the base, and the cooling base is disposed on the thermal insulation base.
In one embodiment, the propulsion assembly includes: the sample platform is positioned in the mounting hole at least partially; the connecting piece is arranged in the mounting cavity and extends into the mounting hole to be connected with the sample table; the second driving piece is connected with the connecting piece and used for driving the connecting piece to axially move along the mounting hole so as to drive the sample table to axially move along the mounting hole to feed the sample.
Compared with the prior art, frozen ultrathin section device simple structure is compact, can integrate in scanning electron microscope, give scanning electron microscope and cut into slices the function of scanning frozen sample simultaneously, specifically, this frozen ultrathin section device passes through propulsion unit, cutting mechanism and temperature regulation subassembly's cooperation, realize that the sample advances and cut into slices in succession in frozen environment, three-dimensional structure characterization for large-size biological sample provides effectual structural support, and this equipment can control cutting mechanism and sample's temperature unanimity, avoid destroying the structure of sample from this, make scanning electron microscope also can implement section imaging technique to frozen sample, and then observe the most true structure of sample, bring very big facility for relevant research work.
Drawings
In order to more clearly illustrate the technical solutions of embodiments or conventional techniques of the present application, the drawings that are required to be used in the description of the embodiments or conventional techniques will be briefly described below, and it is apparent that the drawings in the following description are only some embodiments of the present application, and other drawings may be obtained according to these drawings without inventive effort for a person of ordinary skill in the art.
Fig. 1 is a schematic structural view of a frozen ultrathin section device according to an embodiment of the application.
Fig. 2 is a schematic structural view of the frozen microtome of fig. 1.
Fig. 3 is a partial enlarged view at a in fig. 2.
FIG. 4 is a top view of the frozen microtome of FIG. 1;
FIG. 5 is a cross-sectional view B-B in FIG. 4;
fig. 6 is a C-C cross-sectional view of fig. 4.
Reference numerals: 100. a frozen ultrathin section device; 10. a sample feeding mechanism; 11. a sample stage; 12. a propulsion assembly; 121. a support bracket; 122. a connecting piece; 123. a second driving member; 124. a second bearing; 125. a mounting cavity; 126. a mounting hole; 20. a cutting mechanism; 21. a tool holder; 211. a connecting frame; 212. a heat insulation frame; 22. a heat insulating block; 23. a knife slot; 24. a cutting blade; 241. a knife handle; 242. a cutter head; 30. a driving mechanism; 31. a slide block; 32. a screw rod; 33. a first driving member; 34. a screw nut; 35. a support base; 36. a first bearing; 37. a guide rail; 40. a temperature adjusting mechanism; 41. cooling the base; 42. a cooling pipeline; 43. a flexible heat conducting strip; 44. a heat insulation seat; 45. a first temperature monitor; 46. a second temperature monitor; 47. an elastic member; 50. a base; 200. and (3) a sample.
Detailed Description
In order to make the above objects, features and advantages of the present application more comprehensible, embodiments accompanied with figures are described in detail below. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present application. This application is, however, susceptible of embodiment in many other forms than those described herein and similar modifications can be made by those skilled in the art without departing from the spirit of the application, and therefore the application is not to be limited to the specific embodiments disclosed below.
It will be understood that when an element is referred to as being "mounted" or "disposed" on another element, it can be directly on the other element or intervening elements may also be present. When a component is considered to be "connected" to another component, it can be directly connected to the other component or intervening components may also be present. The terms "vertical," "horizontal," "upper," "lower," "left," "right," and the like are used in the description of the present application for purposes of illustration only and do not represent the only embodiment.
Furthermore, the terms "first," "second," and the like, are used for descriptive purposes only and are not to be construed as indicating or implying a relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defining "a first" or "a second" may explicitly or implicitly include at least one such feature. In the description of the present application, the meaning of "plurality" is at least two, such as two, three, etc., unless explicitly defined otherwise.
In this application, unless expressly stated or limited otherwise, a first feature "up" or "down" a second feature may be a direct contact of the first feature with the second feature, or an indirect contact of the first feature with the second feature via an intervening medium. Moreover, a first feature being "above," "over" and "on" a second feature may be a first feature being directly above or obliquely above the second feature, or simply indicating that the first feature is higher in level than the second feature. The first feature being "under", "below" and "beneath" the second feature may be the first feature being directly under or obliquely under the second feature, or simply indicating that the first feature is less level than the second feature.
Unless defined otherwise, all technical and scientific terms used in the specification of this application have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used in the description of the present application is for the purpose of describing particular embodiments only and is not intended to be limiting of the present application. The term "and/or" as used in the specification of this application includes any and all combinations of one or more of the associated listed items.
Referring to fig. 1 to 6, an embodiment of the present application provides a frozen microtome 100 that can be integrated into a scanning electron microscope, wherein the frozen microtome 100 includes a base 50, a sample feeding mechanism 10, a cutting mechanism 20, a driving mechanism 30, and a temperature adjusting mechanism 40; the sample feeding mechanism 10 is arranged on the base 50, wherein the cutting mechanism 20 is used for cutting the sample 200 so as to enable the sample 200 to have a section, so that the scanning electron microscope can timely scan the section, the driving mechanism 30 is arranged on the base 50 and can move relative to the base 50, and the driving mechanism 30 is connected with the cutting mechanism 20 and is used for driving the cutting mechanism 20 to move and continuously cut the sample 200; the sample feeding mechanism 10 comprises a sample table 11 and a pushing component 12, wherein the sample table 11 is used for supporting a sample 200 to be scanned, the pushing component 12 is connected with the sample table 11 and is used for pushing the sample table 11 to move and feed the sample 200, so that the sample 200 can be fed while cutting, the sample feeding mechanism can timely scan after a section is cut, the section structure is prevented from being damaged, and the three-dimensional structure of the sample 200 is constructed; the temperature adjusting mechanism 40 is in heat conduction connection with the sample table 11 and the cutting mechanism 20 respectively, and is used for adjusting the temperature of the sample 200 and the temperature of the cutting mechanism 20 to be consistent, avoiding the damage to the structure of the section when cutting due to temperature difference, and further providing the high-quality section of the sample 200 for the scanning electron microscope.
It can be appreciated that the frozen ultrathin section device 100 achieves feeding and continuous section of the sample 200 in a frozen environment through the cooperation of the pushing component 12, the cutting mechanism 20 and the temperature adjusting component, so as to realize sequential section imaging, provide effective structural support for three-dimensional structural characterization of the large-size biological sample 200, and the device can control the cutting mechanism 20 to be consistent with the temperature of the sample 200, thereby avoiding damaging the structure of the sample 200, enabling the scanning electron microscope to implement section imaging technology on the frozen sample 200, further observing the truest structure of the sample 200, and bringing great convenience to related research work. In addition, the sample 200 can be directly scanned without moving after being sliced, so that the damage to the section of the sample 200 is effectively avoided, the truest structure of the sample 200 is further ensured, the base 50, the sample feeding mechanism 10, the cutting mechanism 20, the driving mechanism 30 and the temperature regulating mechanism 40 are simple and compact in structure and can be integrated in a scanning electron microscope, and the industrial problem that equipment for imaging the sequence section of the frozen sample 200 in the scanning electron microscope is lacking at present is effectively solved.
As shown in fig. 1 to 3, the cutting mechanism 20 includes a cutter holder 21 and a cutter 24, and the cutter holder 21 is connected to a driving mechanism 30 and is movable relative to the sample stage 11; the cutting blade 24 is fixedly connected with the tool rest 21 and cuts the sample 200 on the sample table 11 when moving along with the tool rest 21, wherein the cutting blade 24 is in heat conduction connection with the temperature regulating mechanism 40, so that the temperature of the cutting blade 24 is regulated by the temperature regulating mechanism 40 to be consistent with the temperature of the sample 200.
In one embodiment, as shown in fig. 2 and 3, the tool holder 21 includes a connection frame 211 and a heat insulating frame 212, and the connection frame 211 is connected to the driving mechanism 30 and is movable relative to the sample stage 11; the heat insulation frame 212 is connected with the connecting frame 211 and the cutting blade 24 respectively and is used for blocking heat conduction between the connecting frame 211 and the cutting blade 24. By the arrangement of the cutter rest 21, heat transfer from the cutter 24 to the connecting frame 211 and the driving mechanism 30 can be blocked, heat loss is avoided, and the efficiency of temperature regulation is improved.
Wherein the connecting frame 211 is of a solid structure, thereby improving the rigidity of the cutting mechanism 20 and ensuring the cutting precision.
In one embodiment, the insulation frame 212 is preferably formed from a relatively strong, relatively low thermal conductivity polyether ether ketone material.
Further, referring to fig. 3, the cutting mechanism 20 further includes a heat insulation block 22, where the heat insulation block 22 is disposed between the cutting blade 24 and the heat insulation frame 212, for enhancing heat conduction between the heat insulation frame 212 and the cutting blade 24, and effectively preventing the cutting blade 24 and other structures from heat conduction with each other through multi-layer heat insulation.
With continued reference to fig. 3, the heat insulation block 22 is provided with a cutter groove 23, the cutting blade 24 is disposed in the cutter groove 23, and the groove bottom and the groove walls on both sides in the cutter groove 23 play a role in three-side heat insulation of the cutting blade 24, so that heat conduction of the cutting blade 24 to other structures and to the environment is effectively avoided. Of course, in other embodiments, the knife slot 23 may be formed in the insulation frame 212, and the insulation block 22 is disposed in the knife slot 23.
As shown in fig. 3, the cutting blade 24 includes a blade handle 241 and a blade head 242, and the blade handle 241 is fixedly connected with the blade holder 21 and is thermally connected with the temperature adjusting mechanism 40; the cutter head 242 is fixed to the cutter handle 241, and the cutter head 242 can cut the sample 200 on the sample stage 11 when moving along with the cutter holder 21. Specifically, the holder 241 is mounted in the pocket 23, and is capable of reciprocating along the pocket 23 toward the sample 200 to perform a cutting operation.
As shown in fig. 1, 2, 4 and 5, the driving mechanism 30 includes a slider 31, a screw 32 and a first driving member 33, the slider 31 is provided on the base 50 and is movable relative to the base 50, and the cutting mechanism 20 is provided on the slider 31 and is connected to the slider 31; the screw rod 32 is in transmission connection with the sliding block 31 and is used for driving the sliding block 31 to slide relative to the base 50; the first driving member 33 is used for driving the screw 32 to move. The rotary motion of the screw rod 32 is converted into the translational motion of the sliding block 31, so that the cutting mechanism 20 is driven to move smoothly and the sample 200 is cut, precise cutting action can be realized, and the thickness of the slice is controlled. Of course, in other embodiments, the specific structure of the driving mechanism 30 is not limited to that described above or shown in the drawings, and may be, for example, a cylinder driving, a rack and pinion driving, a belt driving, or the like.
In one embodiment, the first drive 33 is configured as a motor that enables high-speed cutting movement of the cutting blade 24, thereby ensuring an ultra-thin thickness of the slice and flatness of the slice surface, providing a high quality sample 200 surface for further scanning electron microscope characterization.
As shown in fig. 5, the driving mechanism 30 further includes a screw nut 34, the screw nut 34 is in threaded connection with the screw rod 32, the screw nut 34 is fixedly connected with the slider 31, and when the screw rod 32 rotates, the screw nut 34 is driven to reciprocate along the screw rod 32, so as to drive the slider 31 and the cutting mechanism 20 to reciprocate for cutting.
As shown in fig. 5, the driving mechanism 30 further includes a support seat 35 and a first bearing 36, the support seat 35 is used for supporting the screw rod 32 to make the screw rod 32 move stably, the first bearing 36 is disposed on the support seat 35, and the screw rod 32 is disposed through the first bearing 36, so that the screw rod 32 can rotate through the first bearing 36.
With continued reference to fig. 5, at least two support seats 35 are provided, wherein two support seats 35 are respectively provided at two ends of the screw rod 32 to enable the screw rod 32 to move smoothly, and of course, in other embodiments, the number of the support seats 35 is not limited, for example, one, two, three, four, etc. are provided.
As shown in fig. 4, the driving mechanism 30 further includes a guide rail 37, and the guide rail 37 is provided on the base 50 and corresponds to the slider 31, and the slider 31 can slide along the guide rail 37 to ensure reliability and smoothness of movement.
As shown in fig. 1 to 4 and 6, the temperature adjusting mechanism 40 includes a cooling base 41, a cooling device (not shown), and a cooling pipe 42, wherein the cooling base 41 is thermally connected with the cutting mechanism 20 for adjusting the temperature of the cutting mechanism 20; the cooling device is used for providing a refrigerant and refrigerating; the cooling pipeline 42 is at least partially arranged in the cooling base 41 and the sample table 11, and a refrigerant conveyed by the cooling equipment flows through the cooling pipeline 42 to adjust the temperature of the cooling base 41 and the sample table 11.
Specifically, as shown in fig. 1 and 3, the cooling base 41 and the sample stage 11 are each provided with at least one cooling pipe 42, and the cooling pipe 42 penetrates through the corresponding cooling base 41 and sample stage 11, so that the cooling medium is output from the cooling device into the cooling pipe 42 and circulated into the cooling device through the cooling pipe 42, and the cooling medium continuously conducts heat to the cooling base 41 and the sample stage 11 through the cooling pipe 42, thereby adjusting the temperatures of the cooling base 41 and the sample stage 11.
As shown in fig. 3, the temperature adjustment mechanism 40 further includes a flexible heat conduction band 43, and the flexible heat conduction band 43 is connected with the cooling base 41 and the cutting mechanism 20 for heat-conducting adjustment of the temperature of the cutting mechanism 20. Specifically, the flexible heat conduction band 43 is connected with the knife handle 241, can directly conduct heat to the knife handle 241 and the knife head 242, and temperature regulation speed is faster, and through the setting of flexible heat conduction band 43, with heat concentrated transfer to the cutting sword 24, be favorable to preventing the temperature heat dissipation to the environment in, and flexible heat conduction band 43 possesses flexible elasticity, so does not influence the motion and the section of cutting sword 24. In other embodiments, the temperature of the cutting blade 24 may be adjusted by other means, such as providing a cooling circuit 42 directly on the thermally insulated housing 212 for adjusting the temperature of the thermally insulated housing 212, and further thermally conducting the heat with the cutting blade 24 through the thermally insulated housing 212.
With continued reference to fig. 3, the temperature adjustment mechanism 40 further includes a heat insulation seat 44, the heat insulation seat 44 is mounted on the base 50, and the cooling base 41 is disposed on the heat insulation seat 44 to prevent the cooling base 41 from conducting heat with other structures such as the base 50.
As shown in fig. 3, the frozen ultrathin section device 100 further comprises a control system (not shown), the temperature adjusting mechanism 40 further comprises a first temperature monitor 45 and a second temperature monitor 46, the first temperature monitor 45 is used for monitoring the temperature of the cutting mechanism 20 in real time so as to adjust the temperature of the cutting mechanism 20, the second temperature monitor 46 is used for monitoring the temperature of the sample stage 11 in real time so as to adjust the temperature of the sample stage 11, and further adjust the temperature of the sample 200, so that the phenomenon of melting caused by overhigh temperature of the sample 200 is avoided; the control system is respectively connected with the first temperature monitor 45, the second temperature monitor 46 and the cooling equipment electrically and/or in a signal manner, and is used for controlling and regulating the amount of the refrigerant conveyed into the cooling pipeline 42 by the cooling equipment in real time according to the temperature data fed back by the first temperature monitor 45 and the second temperature monitor 46 and the preset target temperature contrast in the control system, so that the temperature of the cutting blade 24 is regulated to be consistent with the temperature of the sample 200, and the temperature is consistent with the preset target temperature, and the damage to the sample 200 caused by the temperature difference when the cutting blade 24 cuts the sample 200 is prevented. Specifically, when the frozen ultrathin section device 100 is integrated in a scanning electron microscope, the cooling equipment is arranged outside the scanning electron microscope, and the cooling pipeline 42 is communicated with the cooling equipment through a flange interface of the scanning electron microscope.
In one embodiment, the first temperature monitor 45 is provided as a temperature sensor for measuring the temperature of the cutting blade 24 in real time.
As shown in fig. 6, the temperature adjusting mechanism 40 further includes an elastic member 47, where the first temperature monitor 45 is disposed in the cutter slot 23 and between the cutter slot 23 and the cutter 24, and the elastic member 47 is disposed between the first temperature monitor 45 and the cutter slot 23, and the elastic force of the elastic member 47 makes the first temperature monitor 45 closely adhere to the cutter 24 all the time, so that the temperature of the cutter 24 can be measured more accurately.
In one embodiment, the elastic member 47 is provided as a compression spring, however, in other embodiments, the elastic member 47 may be provided as other elastic structures.
In one embodiment, the second temperature monitor 46 is a temperature sensor, the through hole is formed in the sample stage 11, and the temperature sensor is disposed in the through hole, so that the temperature of the sample stage 11 can be measured more accurately.
As shown in fig. 2, 4 and 6, the pushing assembly 12 includes a bearing seat 121, a connecting piece 122 and a second driving piece 123, wherein a mounting hole 126 and a mounting cavity 125 which are mutually communicated are formed in the bearing seat 121, and the sample stage 11 is at least partially positioned in the mounting hole 126; the connecting piece 122 is arranged in the mounting cavity 125 and extends into the mounting hole 126 to be connected with the sample table 11; the second driving member 123 is connected to the connecting member 122, and is used for driving the connecting member 122 to move axially along the mounting hole 126, so as to drive the sample stage 11 to move axially along the mounting hole 126 to feed the sample 200. The guiding of the movement of the sample stage 11 is realized through the mounting holes 126, so that the movement of the sample stage 11 is prevented from being deviated. In addition, the connecting piece 122 and the second driving piece 123 are arranged in the mounting cavity 125, which is beneficial to protecting the safety of the structure.
As shown in fig. 2, the support base 121 is mounted on the base 50, the cooling base 41 is mounted on the support base 121, the cutting blade 24 is located above the support base 121, and the driving mechanism 30 is disposed parallel to the support base 121, so that the structural layout of the frozen ultrathin slice apparatus 100 is compact.
In one embodiment, the second driving member 123 is a piezoelectric driver, and the high-precision motion of the piezoelectric driver can obtain the slice thickness of the nanometer level, so that the Z-direction resolution of the sample 200 during three-dimensional characterization is practically ensured. Meanwhile, continuous slicing of the millimeter-sized sample 200 can be realized through continuous feeding of the piezoelectric driver, effective platform support is provided for three-dimensional structural representation of the large-size biological sample 200, and on the other hand, cutting motion is completed through driving the cutting blade 24 by combining the high-speed characteristic of the driving motor of the cutting blade 24, so that flatness of the surface obtained by slicing is ensured, and high-quality sample surface is provided for further scanning electron microscope representation.
The connecting member 122 is a piezoelectric insulating rod, so that heat conduction between the sample stage 11 and the second driving member 123 can be avoided.
As shown in fig. 6, the pushing assembly 12 further includes a second bearing 124, where the second bearing 124 is disposed in the mounting hole 126 and between the connecting member 122 and the wall of the mounting hole 126, and the sample stage 11 and the connecting member 122 can reduce friction with the mounting hole 126 through the second bearing 124, so that the feeding action of the sample stage 11 is reliable, sensitive and smooth, and is particularly suitable for obtaining a slice thickness of nanometer scale.
The control system is electrically and/or signally connected to the first driving member 33 and the second driving member 123, so that the movement of the cutter 24 and the feeding amount of the sample 200 can be comprehensively controlled, thereby precisely controlling the thickness of the cut and realizing automatic and precise slicing.
The technical features of the above-described embodiments may be arbitrarily combined, and all possible combinations of the technical features in the above-described embodiments are not described for brevity of description, however, as long as there is no contradiction between the combinations of the technical features, they should be considered as the scope of the description.
The above examples only represent a few embodiments of the present application, which are described in more detail and are not to be construed as limiting the scope of the claims. It should be noted that it would be apparent to those skilled in the art that various modifications and improvements could be made without departing from the spirit of the present application, which would be within the scope of the present application. Accordingly, the scope of the present application is to be determined by the following claims.
Claims (11)
1. A frozen microtome device capable of being integrated into a scanning electron microscope, the frozen microtome device comprising:
a base (50);
the sample feeding mechanism (10) is arranged on the base (50), the sample feeding mechanism (10) comprises a sample table (11) and a pushing assembly (12), the sample table (11) is used for supporting a sample (200) to be scanned, and the pushing assembly (12) is connected with the sample table (11) and is used for pushing the sample table (11) to move and feed the sample (200);
a cutting mechanism (20) for cutting the sample (200) so that the sample (200) has a cross section;
the driving mechanism (30) is arranged on the base (50) and can move relative to the base (50), and the driving mechanism (30) is connected with the cutting mechanism (20) and is used for driving the cutting mechanism (20) to move and cut a sample (200);
and the temperature adjusting mechanism (40) is respectively in heat conduction connection with the sample table (11) and the cutting mechanism (20) and is used for adjusting the temperature of the sample (200) and the temperature of the cutting mechanism (20).
2. Frozen microtome according to claim 1, characterized in that the cutting mechanism (20) comprises:
a knife rest (21) connected to the drive mechanism (30) and movable relative to the sample stage (11);
and the cutting knife (24) is fixedly connected with the knife rest (21) and can cut the sample (200) on the sample table (11) along with the movement of the knife rest (21), wherein the cutting knife (24) is in heat conduction connection with the temperature regulating mechanism (40).
3. Frozen microtome according to claim 2, characterized in that the knife holder (21) comprises:
a link (211) connected to the drive mechanism (30) and movable relative to the sample stage (11);
and the heat insulation frame (212) is respectively connected with the connecting frame (211) and the cutting knife (24) and is used for blocking heat conduction between the connecting frame (211) and the cutting knife (24).
4. A frozen microtome according to claim 3, characterized in that the cutting mechanism (20) further comprises a heat insulating block (22), the heat insulating block (22) being arranged between the cutting blade (24) and the heat insulating frame (212) for blocking heat conduction between the heat insulating frame (212) and the cutting blade (24).
5. The frozen microtome according to claim 2, wherein the cutting blade (24) comprises:
the knife handle (241) is fixedly connected with the knife rest (21) and is in heat conduction connection with the temperature adjusting mechanism (40);
and a cutter head (242) fixed on the cutter handle (241), wherein the cutter head (242) can cut the sample (200) on the sample stage (11) when moving along with the cutter rest (21).
6. Frozen microtome according to claim 1, characterized in that the drive mechanism (30) comprises:
a slider (31) provided on the base (50) and movable relative to the base (50), the cutting mechanism (20) being provided on the slider (31) and connected to the slider (31);
the screw rod (32) is in transmission connection with the sliding block (31) and is used for driving the sliding block (31) to slide relative to the base (50);
and the first driving piece (33) is used for driving the screw rod (32) to move.
7. Frozen microtome according to claim 1, characterized in that the temperature adjustment mechanism (40) comprises:
a cooling base (41) thermally connected to the cutting mechanism (20) for adjusting the temperature of the cutting mechanism (20);
the cooling device is used for providing a refrigerant and refrigerating;
and a cooling pipeline (42) at least partially arranged in the cooling base (41) and the sample table (11), wherein a refrigerant conveyed by the cooling equipment circulates in the cooling pipeline (42) so as to regulate the temperatures of the cooling base (41) and the sample table (11).
8. The frozen microtome according to claim 7, characterized in that the temperature adjustment mechanism (40) further comprises a flexible heat conducting strip (43), the flexible heat conducting strip (43) being connected with the cooling base (41) and the cutting mechanism (20) for heat conducting cooling of the cutting mechanism (20).
9. The frozen microtome according to claim 7, wherein the temperature adjustment mechanism (40) further comprises:
a first temperature monitor (45) for monitoring the temperature of the cutting mechanism (20) in real time;
a second temperature monitor (46) for monitoring the temperature of the sample stage (11) in real time;
and the control system is respectively connected with the first temperature monitor (45), the second temperature monitor (46) and the cooling equipment electrically and/or in a signal manner, and the control system controls and regulates the quantity of the refrigerant which is conveyed into the cooling pipeline (42) by the cooling equipment in real time according to the temperature data of the first temperature monitor (45) and the second temperature monitor (46).
10. The frozen microtome according to claim 7, wherein the temperature adjustment mechanism (40) further comprises a thermal insulation seat (44), the thermal insulation seat (44) being mounted on the base (50), the cooling base (41) being provided on the thermal insulation seat (44).
11. Frozen microtome according to claim 1, characterized in that the propulsion assembly (12) comprises:
the sample stage comprises a bearing seat (121), wherein a mounting hole (126) and a mounting cavity (125) which are communicated with each other are formed in the bearing seat (121), and the sample stage (11) is at least partially positioned in the mounting hole (126);
the connecting piece (122) is arranged in the mounting cavity (125) and extends into the mounting hole (126) to be connected with the sample table (11);
and the second driving piece (123) is connected with the connecting piece (122) and is used for driving the connecting piece (122) to axially move along the mounting hole (126) so as to drive the sample stage (11) to axially move along the mounting hole (126) to feed the sample (200).
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211727177.2A CN116448795A (en) | 2022-12-30 | 2022-12-30 | Frozen ultrathin section device capable of being integrated in scanning electron microscope |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202211727177.2A CN116448795A (en) | 2022-12-30 | 2022-12-30 | Frozen ultrathin section device capable of being integrated in scanning electron microscope |
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| CN116448795A true CN116448795A (en) | 2023-07-18 |
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| CN202211727177.2A Pending CN116448795A (en) | 2022-12-30 | 2022-12-30 | Frozen ultrathin section device capable of being integrated in scanning electron microscope |
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