Ultra-precise biological slicer and nano slicing method

Abstract

The invention discloses an ultraprecise biological slicer and a nanometer slicing method, and relates to the technical field of machine.The ultraprecise biological slicer comprises a base, a marble platform is installed on the base through air vibration isolation, hydrostatic pressure guide rails are installed on the two sides of the marble platform, and a gantry support is installed between the two hydrostatic pressure guide rails; a groove is formed in the top of the gantry support, a knife rest is arranged in the groove, a blade is installed on the knife rest, a mechanical lifting table is arranged on the marble below the blade, a two-dimensional adjusting table is installed on the mechanical lifting table, a sample clamp is installed on the two-dimensional adjusting table, and the sample clamp is installed on the mechanical lifting table. A microscope adjusting table is further arranged on the marble platform, a telecentric microscope is mounted on the microscope adjusting table, and the telecentric microscope faces the upper surface of the sample at the sample clamp. According to the invention, the stability of the ultrathin section of the biological sample is guaranteed, and the influence of cutter wear on the slicing effect is avoided.

Description

Technical Field

[0001] This invention relates to the field of mechanical technology, specifically to an ultra-precision biological slicer and a nano-slicing method. Background Technology

[0002] In the field of pathological research, biomicrographs play a crucial role, enabling researchers to observe detailed information about tissue structures using electron microscopes, thereby identifying pathological changes and diagnosing diseases. As pathological research continues to advance, the performance requirements for biomicrographs are gradually increasing. Currently, researchers need to be able to reliably cut biological sample sections with a thickness of less than 1 μm to adapt to the development of high-resolution microscopic imaging technology.

[0003] However, existing biological microtome machines still have certain limitations in performance. The main problem lies in the limited precision of the feed axis, which cannot achieve single-step feed rates below 1 μm, and the linearity of the slicing axis restricts the minimum thickness of the sample slices. Within the effective stroke of the device, the linearity of the slicing axis is >1 μm. When the slice thickness is set below 1 μm, significant problems such as uneven slice thickness, missed slices, over-slicing, and discontinuous slices occur. Furthermore, the structure of existing biological microtome machines also suffers from insufficient rigidity. Structural deformation during sample slicing also causes uneven slice thickness, missed slices, over-slicing, and discontinuous slices. During sample trimming, setting a large feed rate results in significant force on the cutting tool, leading to severe tool wear after trimming and affecting the quality of the slices.

[0004] Therefore, those skilled in the art are dedicated to developing a new type of ultra-precision biological slicer to ensure the stability of ultrathin sections of biological samples and to avoid the impact of blade wear on the slicing effect. Summary of the Invention

[0005] In view of the above-mentioned deficiencies of the prior art, the technical problem to be solved by the present invention is to provide a novel ultra-precision biological slicer to ensure the stability of ultra-thin sections of biological samples and to avoid the impact of blade wear on the slicing effect.

[0006] To achieve the above objectives, the present invention provides an ultra-precision biological slicer, comprising a base on which a marble platform is mounted via air vibration isolation. Hydrostatic guide rails are mounted on both sides of the marble platform, and a gantry support is installed between the two hydrostatic guide rails. A groove is provided at the top of the gantry support, and a blade holder is disposed within the groove. A blade is mounted on the blade holder. A mechanical lifting platform is disposed on the marble below the blade. A two-dimensional adjustment stage is mounted on the mechanical lifting platform, and a sample holder is mounted on the two-dimensional adjustment stage. A microscope adjustment stage is also disposed on the marble platform, and a telecentric microscope is mounted on the microscope adjustment stage, with the telecentric microscope facing the upper surface of the sample at the sample holder.

[0007] In a preferred embodiment of the present invention, protrusions are provided on both sides of the top of the marble platform, and the hydrostatic guide rails are fitted onto the protrusions on both sides of the marble platform.

[0008] In a preferred embodiment of the present invention, a piezoelectric ceramic is also installed in the groove at the top of the gantry bracket. The piezoelectric ceramic is fixedly connected to the tool holder seat of the tool holder. A tool holder body is installed below the bottom of the tool holder seat, and the blade is interference-clamped in the slender groove at the top of the tool holder body.

[0009] Furthermore, chip collection grooves are fitted on both sides of the tool holder.

[0010] In a preferred embodiment of the present invention, the mechanical lifting platform includes a first fixed seat fixedly installed on a marble platform, a first movable seat slidably disposed above the first fixed seat, a first connecting member and a second connecting member fixedly installed on the inner side wall of the bottom surface and the outer side wall of the top surface of the first movable seat, the first connecting member connecting to a movable platform, and the top surface of the first movable seat and the bottom surface of the movable platform being configured as a set of wedge-shaped surfaces, the first fixed seat and the first movable seat being respectively connected to a linear motor magnetic rail and a coil, the first fixed seat and the movable platform being respectively connected to a guide seat and a guide rod, and the guide rod being installed above the guide seat.

[0011] In a preferred embodiment of the present invention, the two-dimensional adjustment platform includes a second fixed seat fixedly installed on the mechanical lifting platform, a second movable seat slidably disposed above the second fixed seat, a third connecting member fixedly installed on the adjacent side walls of the second movable seat, the third connecting member being connected to a movable rod, the movable rod being threadedly connected to a first threaded ring, and the first threaded ring being fixedly installed on the fixed seat.

[0012] In a preferred embodiment of the present invention, the sample clamp includes a base plate fixedly installed above the top of the two-dimensional adjustment platform, and two inwardly inclined clamping plates are symmetrically fixedly installed on both sides of the base plate, and the sample is placed between the two clamping plates.

[0013] In a preferred embodiment of the present invention, the microscope adjustment stage includes an adjustment stage base fixedly installed above the marble platform. The adjustment stage base is provided with a first manual linear displacement stage, a second manual linear displacement stage and a right-angle connecting plate from bottom to top. A third manual linear displacement stage is provided on the side wall of the right-angle connecting plate. A microscope support is fixedly installed on the third manual linear displacement stage, and the telecentric microscope is installed on the microscope support.

[0014] Preferably, the first manual linear displacement stage moves back and forth, the second manual linear displacement stage moves left and right, and the third manual linear displacement stage moves up and down. The first, second, and third manual linear displacement stages have the same structure, including a rotating rod. The rotating rod is threadedly connected to a second screw ring, and a linear displacement stage is fixedly installed at the other end of the rotating rod. The second screw ring is adjustablely installed on the linear displacement stage by bolts.

[0015] This invention also provides a nanoslicing method, which uses an ultra-precision biological slicer as described above, and includes the following steps:

[0016] S1. Fix the sample in the sample fixture, control the two-dimensional adjustment stage, and move the sample to the repair station;

[0017] S2. Adjust the mechanical lifting platform to bring the sample closer to the blade holder, and adjust the microscope adjustment stage to ensure that the blade and the sample are both within the field of view of the telecentric microscope.

[0018] S3. Start the reciprocating motion of the hydrostatic guide rail, control the mechanical lifting platform to slowly raise the sample holder, and observe the telecentric microscope until the blade contacts the sample surface and chips fall off, then the mechanical lifting platform stops rising.

[0019] S4. Control the mechanical lifting platform to drive the sample clamp upward, with a single step feed of 1-20μm, and feed a total of 5-10 times to complete the sample trimming. Control the two-dimensional adjustment platform to move the sample to the slicing station.

[0020] S5. Control the piezoelectric ceramic to drive the tool holder downward feed, with a single step feed amount of 0.1-0.5μm, feed 1-10 times in total, and collect the sample slices in the chip collection groove to complete the nano-slicing of the sample;

[0021] S6. Control the mechanical lifting platform to move the sample holder away from the knife holder, and the liquid hydrostatic guide rail stops moving.

[0022] Compared with the prior art, the present invention has the following beneficial effects:

[0023] 1. This invention improves the support form of the biological slicer's slicing axis from a mechanical linear guide with micron-level straightness to a hydrostatic guide with 50nm straightness, thereby improving its motion straightness to below 50nm. That is, the motion error of the axis is <50nm, which can be ignored relative to the 1μm slice thickness, thus ensuring the stability of ultrathin biological sample slices.

[0024] 2. This invention improves the drive mechanism of the feed axis of the traditional biological slicer from a micron-level motor with a lead screw and nut to a nanon-level piezoelectric ceramic, thereby increasing the single-step feed accuracy of this component to 50nm. This provides the equipment foundation for the current thickness limit of biological slices and can further contribute to the development of the field of pathological analysis.

[0025] 3. This invention uses two sets of hydrostatic guide rails in the same direction. By increasing the area of ​​the guide rail support surface, the rigidity of the guide rail is increased. The two sets of guide rails are connected by a gantry structure, which makes the structure more stable. Compared with a single hydrostatic guide rail, its rigidity is improved. The gantry structure improves the stability of the machine tool structure.

[0026] 4. This invention adopts a dual-station equipment layout for trimming and slicing. Through a two-dimensional adjustment stage, the sample is moved between the trimming and slicing stations. After trimming, the sample is moved to the slicing station and sliced ​​using a new blade, thus avoiding the impact of blade wear on the slicing effect.

[0027] The following will further explain the concept, specific structure, and technical effects of the present invention in conjunction with the accompanying drawings, so as to fully understand the purpose, features, and effects of the present invention. Attached Figure Description

[0028] Figure 1 A three-dimensional structural schematic diagram of the ultra-precision biological slicer provided by the present invention;

[0029] Figure 2 A front view of the ultra-precision biological slicer provided by the present invention;

[0030] Figure 3 A side view of the ultra-precision biological slicer provided by the present invention;

[0031] Figure 4 A schematic diagram showing the connection relationship between the piezoelectric ceramic, the blade holder, and the blade in the ultra-precision biological slicer provided by this invention;

[0032] Figure 5 This is a schematic diagram of the structure of the two-dimensional adjustment stage in the ultra-precision biological slicer provided by the present invention;

[0033] Figure 6 An exploded view of the microscope adjustment stage in the ultra-precision biological slicer provided by the present invention;

[0034] Figure 7 This is a schematic diagram of the ultra-precision biological slicer provided by the present invention in the trimming station and the slicing station; wherein, the left side is the trimming station and the right side is the slicing station.

[0035] Figure 8 This is a schematic diagram of the mechanical lifting platform in the ultra-precision biological slicer provided by the present invention; Detailed Implementation

[0036] The following description, with reference to the accompanying drawings, illustrates several preferred embodiments of the present invention to make its technical content clearer and easier to understand. The present invention can be embodied in many different forms, and the scope of protection of the present invention is not limited to the embodiments mentioned herein.

[0037] In the accompanying drawings, components with the same structure are indicated by the same numerical designation, and components with similar structures or functions are indicated by similar numerical designations. The dimensions and thicknesses of each component shown in the drawings are arbitrary, and the present invention does not limit the dimensions and thicknesses of each component. To make the illustrations clearer, the thickness of some components has been appropriately exaggerated in the drawings.

[0038] This invention primarily addresses sample preparation before pathological analysis. By improving key precision indicators of the equipment, it breaks through the current limits on sample slice thickness. Specifically, by changing the drive mechanism of the feed axis and the support mechanism of the slice axis, this invention improves the feed accuracy and linearity of the equipment by an order of magnitude, greatly ensuring the stability of ultrathin biological sample slices. Its performance indicators are absolutely leading among similar devices. Based on the necessity of this equipment in the preliminary preparation work for pathological analysis, it is predicted to have practical significance and broad development prospects.

[0039] The following is a description through specific embodiments.

[0040] Example 1

[0041] like Figure 1-3 The ultra-precision biological slicer shown mainly includes: a base 1, an air vibration isolation 2, a marble platform 3, a liquid hydrostatic guide rail 4, a gantry support 5, a mechanical lifting platform 6, a two-dimensional adjustment platform 7, a sample clamp 8, a piezoelectric ceramic 9, a blade holder 10, a chip collection groove 11, a microscope adjustment stage 12, a telecentric microscope 13, a sample 14, and a blade 15.

[0042] The air vibration isolation 2 has its bottom and top surfaces fixedly connected to the upper surface of the base 1 and the bottom surface of the marble platform 3, respectively. The hydrostatic guide rail 4 is fitted onto the protrusions on both sides of the marble platform 3, and its upper surface is fixedly connected to the bottom surface of the gantry bracket 5. The mechanical lifting platform 6, the two-dimensional adjustment platform 7, and the sample clamp 8 are fixedly centered on the upper surface of the marble platform 3 from bottom to top. The sample clamp 8 includes a base plate fixedly installed above the top of the two-dimensional adjustment platform, and two inwardly inclined clamping plates symmetrically fixedly installed on both sides of the base plate. The sample 14 is placed between the two clamping plates.

[0043] The top of the gantry bracket 5 has a groove, and a piezoelectric ceramic 9 is installed in the groove, such as... Figure 4 As shown, the piezoelectric ceramic 9 is fixedly connected to the tool holder 10-2 of the tool holder 10. The tool holder body 10-1 is installed below the bottom of the tool holder 10-2, and the blade 15 is interference-fitted into the elongated groove at the top of the tool holder body 10-1. The chip collection groove 11 is fitted on both sides of the tool holder 10. The telecentric microscope 13 is fixedly mounted on the upper surface of the marble platform 3 via the microscope adjustment stage 12, facing the upper surface of the sample 14.

[0044] The mechanical lifting platform 6 in this invention has the following structure: Figure 8 As shown, the system includes a first fixed seat 6-1 fixedly installed on a marble platform 3. A first movable seat 6-2 is slidably disposed above the first fixed seat 6-1. A first connecting piece 6-4 and a second connecting piece 6-5 are fixedly installed on the inner side wall of the bottom surface and the outer side wall of the top surface of the first movable seat 6-2, respectively. The first connecting piece 6-4 is connected to a movable platform 6-3. The top surface of the first movable seat 6-2 and the bottom surface of the movable platform 6-3 are set as a set of wedge-shaped surfaces. The first fixed seat 6-1 and the first movable seat 6-2 are respectively connected to a linear motor magnetic rail 6-7 and a coil 6-6. The first fixed seat 6-1 and the movable platform 6-3 are respectively connected to a guide seat 6-8 and a guide rod 6-9. The guide rod is installed above the guide seat.

[0045] In this invention, the first movable seat 6-2 is driven by the linear motor magnetic rail 6-7 to move horizontally, and the movable table 6-3 connected to the guide rod 6-9 can only move up and down under the constraint of the guide seat 6-8. Since the top surface of the first movable seat 6-2 and the bottom surface of the movable table 6-3 are set as a set of wedge-shaped surfaces, when the first movable seat 6-2 and the movable table 6-3 move relative to each other in the horizontal direction, the worktable base will rise or fall.

[0046] The structure of the two-dimensional adjustment stage 7 is as follows Figure 5As shown, it includes a second fixed seat 7-1 fixedly installed on the mechanical lifting platform 6, a second movable seat 7-2 slidably arranged above the second fixed seat 7-1, a third connecting piece 7-5 fixedly installed on the adjacent two side walls of the second movable seat 7-2, a moving rod 7-3 connected to the third connecting piece 7-5, a first threaded ring 7-4 threadedly connected to the moving rod 7-3, and the first threaded ring 7-4 fixedly installed on the second fixed seat 7-1.

[0047] By rotating the moving rod 7-3, the moving rod 7-3 can be controlled to move forward, backward, left, or right, thereby driving the second moving seat 7-2 to move through the third connecting piece 7-5, so as to control the position of the sample clamp 8 on the second moving seat 7-2, and thus control the position of the sample 14, which is convenient for trimming or slicing.

[0048] Similarly, in this invention, such as Figure 6 As shown, the microscope adjustment stage 12 includes an adjustment stage base 12-1 fixedly installed above the marble platform 3. From bottom to top, the adjustment stage base 12-1 is provided with a first manual linear displacement stage 12-2, a second manual linear displacement stage 12-3, and a right-angle connecting plate 12-4. A third manual linear displacement stage 12-5 is provided on the side wall of the right-angle connecting plate 12-4. A microscope support 12-6 is fixedly installed on the third manual linear displacement stage 12-5, and a telecentric microscope 13 is installed on the microscope support 12-6. The first manual linear displacement stage 12-2 moves back and forth, the second manual linear displacement stage 12-3 moves left and right, and the third manual linear displacement stage 12-5 moves up and down. In this invention, the first manual linear displacement stage 12-2, the second manual linear displacement stage 12-3, and the third manual linear displacement stage 12-5 have the same structure, including a rotating rod 12-7. The rotating rod 12-7 is threadedly connected to the second screw ring 12-8. The other end of the rotating rod 12-7 is fixedly mounted with a linear displacement stage. The second screw ring 12-8 is adjustablely mounted on the linear displacement stage by bolts.

[0049] By controlling the rotation of the rotating rod 12-7 on the second screw ring 12-8, the movement of the first manual linear displacement stage 12-2, the second manual linear displacement stage 12-3, or the third manual linear displacement stage 12-5 can be adjusted. After adjusting to the appropriate position, the second screw ring 12-8 is fixed to the linear displacement stage with bolts, thereby fixing the position of the linear displacement stage for observation by the telecentric microscope 13.

[0050] Example 2

[0051] The method for nanoslicing using the ultra-precision biological slicer of Example 1 includes the following steps:

[0052] S1. Secure the sample in the sample holder, and control the two-dimensional adjustment stage to move the sample to the trimming station, such as... Figure 7As shown in the left-hand view;

[0053] S2. Adjust the mechanical lifting platform to bring the sample closer to the blade holder, and adjust the microscope adjustment stage to ensure that the blade and the sample are both within the field of view of the telecentric microscope.

[0054] S3. Start the reciprocating motion of the hydrostatic guide rail, control the mechanical lifting platform to slowly raise the sample holder, and observe the telecentric microscope until the blade contacts the sample surface and chips fall off, then the mechanical lifting platform stops rising.

[0055] S4. Control the mechanical lifting platform to move the sample holder upwards, with a single-step feed of 1-20μm, for a total of 5-10 feeds, completing the sample trimming. Then control the two-dimensional adjustment stage to move the sample to the slicing station. Figure 7 As shown in the right-hand view;

[0056] S5. Control the piezoelectric ceramic to drive the tool holder downward feed, with a single step feed amount of 0.1-0.5μm, feed 1-10 times in total, and collect the sample slices in the chip collection groove to complete the nano-slicing of the sample;

[0057] S6. Control the mechanical lifting platform to move the sample holder away from the knife holder, and the liquid hydrostatic guide rail stops moving.

[0058] The preferred embodiments of the present invention have been described in detail above. It should be understood that those skilled in the art can make numerous modifications and variations based on the concept of the present invention without creative effort. Therefore, all technical solutions that can be obtained by those skilled in the art based on the concept of the present invention through logical analysis, reasoning, or limited experimentation on the basis of existing technology should be within the scope of protection defined by the claims.

Claims

1. An ultra-precision biological slicer, characterized in that, The device includes a base on which a marble platform is mounted via air vibration isolation. Hydrostatic guide rails are mounted on both sides of the marble platform, and a gantry support is installed between the two hydrostatic guide rails. A groove is provided at the top of the gantry support, and a knife holder is placed within the groove. A blade is mounted on the knife holder. A mechanical lifting platform is located on the marble below the blade. A two-dimensional adjustment platform is mounted on the mechanical lifting platform, and a sample clamp is mounted on the two-dimensional adjustment platform. A microscope adjustment platform is also provided on the marble platform, and a telecentric microscope is mounted on the microscope adjustment platform, facing the upper surface of the sample at the sample clamp.

2. The ultra-precision biological slicer according to claim 1, characterized in that, The marble platform has protrusions on both sides of its top, and the hydrostatic guide rails are fitted onto the protrusions on both sides of the marble platform.

3. The ultra-precision biological slicer according to claim 1, characterized in that, A piezoelectric ceramic is also installed in the groove at the top of the gantry bracket. The piezoelectric ceramic is fixedly connected to the tool holder seat of the tool holder. The tool holder body is installed below the bottom of the tool holder seat, and the blade is interference-clamped in the slender groove at the top of the tool holder body.

4. The ultra-precision biological slicer according to claim 3, characterized in that, The tool holder is also fitted with chip collection grooves on both sides.

5. The ultra-precision biological slicer according to claim 1, characterized in that, The mechanical lifting platform includes a first fixed seat fixedly installed on a marble platform, a first movable seat slidably disposed above the first fixed seat, a first connector and a second connector fixedly installed on the inner side wall of the bottom surface and the outer side wall of the top surface of the first movable seat, the first connector being connected to a movable platform, and the top surface of the first movable seat and the bottom surface of the movable platform being configured as a set of wedge-shaped surfaces, the first fixed seat and the first movable seat being respectively connected to a linear motor magnetic rail and a coil, the first fixed seat and the movable platform being respectively connected to a guide seat and a guide rod, and the guide rod being installed above the guide seat.

6. The ultra-precision biological slicer according to claim 1, characterized in that, The two-dimensional adjustment platform includes a second fixed seat fixedly installed on the mechanical lifting platform. A second movable seat is slidably arranged above the second fixed seat. A third connecting member is fixedly installed on the adjacent side walls of the second movable seat. The third connecting member is connected to a movable rod. The movable rod is threadedly connected to a first threaded ring. The first threaded ring is fixedly installed on the fixed seat.

7. The ultra-precision biological slicer according to claim 1, characterized in that, The sample clamp includes a base plate fixedly installed above the top of the two-dimensional adjustment platform, and two inwardly inclined clamping plates are symmetrically fixedly installed on both sides of the base plate, with the sample placed between the two clamping plates.

8. The ultra-precision biological slicer according to claim 1, characterized in that, The microscope adjustment stage includes an adjustment stage base fixedly installed above the marble platform. From bottom to top, the adjustment stage base is provided with a first manual linear displacement stage, a second manual linear displacement stage and a right-angle connecting plate. A third manual linear displacement stage is provided on the side wall of the right-angle connecting plate. A microscope support is fixedly installed on the third manual linear displacement stage, and the telecentric microscope is installed on the microscope support.

9. The ultra-precision biological slicer according to claim 8, characterized in that, The first manual linear displacement table moves back and forth, the second manual linear displacement table moves left and right, and the third manual linear displacement table moves up and down. The first, second, and third manual linear displacement tables have the same structure, including a rotating rod. The rotating rod is threadedly connected to a second screw ring. A linear displacement table is fixedly installed at the other end of the rotating rod. The second screw ring is adjustablely installed on the linear displacement table by bolts.

10. A method for nanoslicing, characterized in that, Sectioning using the ultra-precision biological slicer as described in any one of claims 1-9 includes the following steps: S1. Fix the sample in the sample fixture, control the two-dimensional adjustment stage, and move the sample to the repair station; S2. Adjust the mechanical lifting platform to bring the sample closer to the blade holder, and adjust the microscope adjustment stage to ensure that the blade and the sample are both within the field of view of the telecentric microscope. S3. Start the reciprocating motion of the hydrostatic guide rail, control the mechanical lifting platform to slowly raise the sample holder, and observe the telecentric microscope until the blade contacts the sample surface and chips fall off, then the mechanical lifting platform stops rising. S4. Control the mechanical lifting platform to drive the sample clamp upward, with a single step feed of 1-20μm, and feed a total of 5-10 times to complete the sample trimming. Control the two-dimensional adjustment platform to move the sample to the slicing station. S5. Control the piezoelectric ceramic to drive the tool holder downward feed, with a single step feed amount of 0.1-0.5μm, feed 1-10 times in total, and collect the sample slices in the chip collection groove to complete the nano-slicing of the sample; S6. Control the mechanical lifting platform to move the sample holder away from the knife holder, and the liquid hydrostatic guide rail stops moving.

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