Ribbon-shaped serial ultrathin section collection tool for transmission electron microscopes

Abstract

To provide a recovery jig for easily, accurately, and quickly recovering ribbon form continuous ultrathin slices (13) floating on a water surface (14) of a knife boat (15) in cutting order to a slit type grid (16).SOLUTION: A recovery method includes: attaching a main body (10) to a knife boat (15); cutting ribbon form continuous ultrathin slices (13); placing the ribbon form continuous ultrathin slices (13) floating on a water surface (14) into a ring (7) of a recovery tool (12); moving the water surface (14) in a protective recovery hole (4) of the main body (10); bringing the ribbon form continuous ultrathin slices (13) close to piece recovery location (5) of the protective recovery hole (4) using lashes (17) or the like; and subjecting the ribbon form continuous ultrathin slices (13) to a pressing method to recover a slit type grid (16).SELECTED DRAWING: Figure 6

Description

[Technical Field]

[0001] The present invention relates to a jig used for producing serial ultrathin sections as a pretreatment for three-dimensional observation using an electron microscope. [Background technology]

[0002] In the serial section cutting method for transmission electron microscopes, sections are connected one by one with an ultramicrotome to form 2 mm long ribbon-like serial ultrathin sections, which are then collected one by one on a slit grid. Due to the limited water surface area of ​​the knife boat, a maximum of seven ribbon-shaped ultrathin sections are prepared. Seven protective loops are used to surround each of the ribbon-shaped serial ultrathin sections, and the handles are hung on the edge of the knife board. Then, a recovery loop is pressed onto the protective loop, and the ribbon-shaped serial ultrathin sections are transferred to the recovery loop. The recovery loop is pressed slightly below the water surface, so that the water surface inside the loop becomes convex. The ribbon-shaped serial ultrathin sections are moved to the apex of the convex surface using an eyelash or similar tool, and the slit-shaped grid, held with a scoop, is pressed horizontally onto the ribbon-shaped serial ultrathin sections, aligning them with the direction of the ribbon-shaped serial ultrathin sections. The slit-shaped grid, along with the water droplets on it, is placed on the prepared support film; once it dries, the support film will become one with the ribbon-shaped serial ultrathin sections and the grid. After drying, the sections are then observed using electronic staining or other methods. [Prior art documents]

[0003] [Non-patent literature] [Non-patent literature] Journal of the Japanese Society of Microscopy Microscope Vol.46, No.4 2011 A reliable method for preparing serial ultrathin sections using a new auxiliary tool Summary of the Invention [Problem to be solved by the invention]

[0004] This had the following drawbacks: Because the protective loops are not fixed to the edge of the boat, they are vulnerable to vibrations during operation and tend to overlap. In addition, the ribbon-shaped serial ultrathin sections must be transferred from the protective loop to the recovery loop, and the loop must then be lowered below the water surface, which is complicated and time-consuming and reduces precision. The present invention has been devised to eliminate the above drawbacks. [Means for solving the problem]

[0005] The present invention has been made to solve the above-mentioned problems, and is a tool for attaching a diamond knife to a knife boat (15), ultra-thinly slicing ribbon-shaped serial ultrathin sections (13) floating on the water surface (14), and then using a recovery tool (12) to move the ribbon-shaped serial ultrathin sections (13) along the water surface (14) to a protective recovery hole (4) in the main body (10) in the cutting order, and then using eyelashes (17) or the like to move the ribbon-shaped serial ultrathin sections (13) to a section recovery location (5) in the protective recovery hole (4) and press them against a slit-type grid (15) for recovery, as shown in Figures 1 to 13. It is a very thin rectangular protective recovery disk (1) with rounded corners, and its longitudinal length is long enough to fit inside the diamond knife boat (15). A protective recovery hole (4) having a generally circular shape is arranged in a row in the center of the width of the protective recovery board (1) at equal intervals. The protective recovery hole (4) is sized to accommodate the ring (7) of the recovery tool (12), and a portion of the arc of the protective recovery hole (4) is cut out to provide a linear section recovery area (5) parallel to the longitudinal edge a (9) in which the ribbon-shaped continuous ultrathin section (13) can be placed. A plurality of support wings (2) are provided near both ends of the longitudinal edge a (9) of the protective recovery board (1) so as to discharge the liquid horizontally from the top surface of the protective recovery board (1). The handle part (6) is hung down from the underside of the longitudinal edge a (9) of the protective recovery board (1) at a slight incline towards the center of the knife boat, and at an appropriate point, it is extended parallel to the width direction of the protective recovery board (1) and is attached to the knife boat ( 15 ) bends upward just before the wall and reaches the knife boat ( 15The retriever (12) has a main body (10) that is bent outward at a right angle to form a grip (8) at the tip, and a very thin, approximately Landolt annular ring (7) large enough to accommodate a ribbon-shaped continuous ultrathin section (13), with a handle (20) at one corner. The present invention is a tool for recovering ribbon-shaped serial ultrathin sections (13) for use in a transmission electron microscope, which is constructed as described above. [Effects of the Invention]

[0006] By changing the operating method to attaching the main body (10) to the knife boat (15) before cutting the ribbon-shaped serial ultrathin sections (13), the ribbon-shaped serial ultrathin sections (13) floating on the water surface (14) of the knife boat (15) after cutting can be quietly and quickly moved to the protective recovery hole (4) using the recovery tool (12). Operations such as moving the sections to the section recovery location (5) within the protective recovery hole (4) are also quiet, and even when the ribbon-shaped serial ultrathin sections (13) are recovered at the section recovery location (5) by pressing them against the slit-type grid (16), there is no adverse effect on the ribbon-shaped serial ultrathin sections (13) floating in the adjacent protective recovery hole (4). Compared to conventional methods, this recovery tool is simpler, more accurate, and faster, and far superior to conventional methods in every respect. [Brief explanation of the drawings]

[0007] [Figure 1] FIG. [Figure 2] FIG. [Figure 3] FIG. [Figure 4] FIG. [Figure 5] FIG. 1 is a partially cutaway view of a knife boat equipped with the present invention. [Figure 6] A perspective view of a ribbon-shaped serial ultrathin section (13) being moved by a ring (7) into a protective collection hole (4). [Figure 7] 1 is a perspective view showing the state in which the ribbon-shaped serial ultrathin sections (13) have been moved to the protective recovery hole (4). [Figure 8] FIG. 1 is a perspective view showing the state in which the ring (7) has been separated from the ribbon-shaped serial ultrathin section (13). [Figure 9]This is a perspective view of the ribbon-shaped serial ultrathin sections (13) being moved to the section collection location (5) using an eyelash or the like. [Figure 10] FIG. 1 is a perspective view showing the state in which ribbon-shaped ultrathin sections (13) are collected on a slit-type grid (16). [Figure 11] FIG. 11 is an enlarged perspective view of a part of FIG. [Figure 12] FIG. 10 is a perspective view showing a state in which the grid is placed on the support film. [Figure 13] FIG. 13 is an enlarged perspective view of FIG. DETAILED DESCRIPTION OF THE INVENTION

[0008] Hereinafter, embodiments of the present invention will be described in detail with reference to FIGS. The present invention is a tool for attaching a main body to a knife boat (15) of a diamond knife, cutting the ribbon-shaped serial ultrathin sections (13) floating on the water surface (14) by ultra-thin slicing, and then using a recovery tool (12) to move the ribbon-shaped serial ultrathin sections (13) to the water surface (14) to a protective recovery hole (4) of the main body (10), and then using eyelashes (17) or the like to move the ribbon-shaped serial ultrathin sections (13) to a section recovery location (5) in the protective recovery hole (4) and press them against a slit-type grid (15) to recover them. As shown in Figures 1 to 13, A very thin rectangular protective recovery disk (1) made of stainless steel with rounded corners, the longitudinal length of which is equal to the length of the diamond knife boat ( 15 ) is the length that fits within The protective recovery board (1) has a row of generally circular protective recovery holes (4) arranged at equal intervals in the center of its width.

[0009] The protective recovery hole (4) is large enough to accommodate the ring (7) of the recovery tool (12), and a portion of the arc of the protective recovery hole (4) is cut out to provide a linear section recovery area (5) parallel to the longitudinal edge a (9) in which the ribbon-shaped continuous ultrathin section (13) can be placed. In order to make the top surface of the protective recovery plate (1) the same height as the knife boat edge (3), the plate support blades (2) are provided at multiple locations near both ends of the longitudinal edge a (9) so that they extend horizontally from the top surface. In this case, it is desirable to provide spacing at two points on both ends that divide the longitudinal edge a (9) into six equal parts.

[0010] In order to cut the ribbon-shaped serial ultrathin sections (13), the long edge b (11) of the protective recovery disc (1) was attached to the knife boat ( 15 The handle part (6) for adjusting the height of the water surface (14) of the protective recovery board (1) hangs down from the underside of the longitudinal edge a (9) of the protective recovery board (1) at a slight incline towards the center of the knife boat, and extends parallel to the width direction of the protective recovery board (1) at an appropriate point, and is connected to the knife boat ( 15 ) bends upward just before the wall and reaches the knife boat ( 15 The main body (10) is provided with a grip portion (8) at the end of a right-angle bend outward.

[0011] A handle (20) is attached to one corner of a very thin, stainless steel, approximately Landolt-shaped ring (7) large enough to accommodate the ribbon-shaped serial ultrathin sections (13) floating on the water surface (14) at the tip of a recovery tool (12) that moves the ribbon-shaped serial ultrathin sections (13) floating on the water surface (14) to the protective recovery hole (4).

[0012] The present invention is a ribbon-shaped serial ultrathin section recovery jig for a transmission electron microscope having the above-mentioned configuration. When using this, The grip portion (8) can be used on either the left or right side, but the following description will be given with the grip portion (8) on the left side. As shown in Figures 4 and 5, the grip part (8) of the main body (10) is held with the grip part (8) facing left and attached horizontally to the knife boat (15). Because the main body (10) is wide at the top, the board support wing (2) is placed on the right side of the knife boat edge (3), and the grip part (8) is pushed into the left side of the knife boat edge (3). The grip part side and the longitudinal edge a (9) of the protective recovery board (1) are braced against each other, so the grip part (8) side can be fixed at any height position on the left inner wall of the knife boat (15).

[0013] As shown in Figure 6, to prepare ribbon-shaped serial ultrathin sections (13), water is poured into the knife boat (15) up to the height of the water surface (14) required for cutting. The grip part (8) is moved up and down around the longitudinal edge a (9) as the axis of rotation to change the angle of the protective recovery plate (1) until the longitudinal edge b (11) is at the same height as the water surface (14), and six ribbon-shaped serial ultrathin sections (13) are cut, the number of which is the number of holes in the protective recovery holes (4). As shown in Figure 6, the ribbon-shaped serial ultrathin sections (13) are placed in the loops (7) of the recovery tool (12) and moved horizontally across the water surface (14) to the recovery protection hole (4). At this time, it is desirable to lower the longitudinal edge b (11) with the handle to make it easier for the section to pass through the loops (7) of the recovery tool (12). As shown in Figures 7 and 8, the ring (7) of the recovery device (12) is a protective recovery hole (4), and when the open part of the ring (7) is raised at a slightly higher angle, the ring (7) gently separates from the water surface (14).

[0014] This procedure is repeated to transfer six ribbon-shaped serial ultrathin sections (13) to the protective collection hole (4). As shown in FIG. 9, the ribbon-shaped serial ultrathin sections (13) are moved with the eyelashes to the section collection location (5) of the protective collection hole (4) and aligned in the straight line. As shown in Figures 10 and 11, the prepared slit-shaped grid (16) is grasped with a scoop (17) and pressed horizontally against the ribbon-shaped serial ultrathin sections (13) to recover them. As shown in FIG. 12, it is placed horizontally on a support film (18) stretched over the hole of a support film stand with a drop of water attached thereto. As shown in FIG. 13, when the water droplets on the support film (18) dry, the ribbon-shaped serial ultrathin sections (13) and the support film (20) on the slit-type grid (16) become one unit, and collection is complete. [Explanation of symbols]

[0015] 1 Protected Recovery Disc 2-panel support wing 3 Knife Boat Edge 4 Protective Recovery Hole 5. Section collection location 6 Handle 7 wheels 8 Grip 9 Long edge a 10 Main Unit 11 Long edge b 12 Recovery tool 13 Ribbon-shaped serial ultrathin sections 14 Water surface 15 Knife Boat 16 slit grid 17 Eyelashes 18 Setsuko 19 Support membrane 20 patterns

Claims

[Claim 1] It is a very thin rectangular protective recovery disk (1) with rounded corners, and its longitudinal length is long enough to fit inside the diamond knife boat (15). The protective recovery board (1) has a row of generally circular protective recovery holes (4) arranged at equal intervals in the center of the width of the board, The protective recovery hole (4) is sized to accommodate the ring (7) of the recovery tool (12), and a part of the arc of the protective recovery hole (4) is cut out to provide a linear section recovery area (5) parallel to the longitudinal edge a (9) in which the ribbon-shaped continuous ultrathin section (13) can be accommodated; A plurality of support wings (2) are provided near both ends of the longitudinal edge a (9) of the protective recovery board (1) so as to eject the water horizontally from the top surface of the protective recovery board (1), a main body in which the handle portion (6) hangs down from the underside of the longitudinal edge a (9) of the protective recovery board (1) at a slight incline towards the center of the knife boat, extends at an appropriate point substantially parallel to the width direction of the protective recovery board (1), bends upward just before the wall of the knife boat (15), and bends outward from the knife boat (15) at a position higher than the knife boat edge (3) at a substantially right angle, at which point the grip portion (8) is provided; The ribbon-shaped serial ultrathin section recovery tool for a transmission electron microscope comprises a recovery tool (12) having a handle (20) at one corner of an extremely thin, approximately Landolt annular ring (7) large enough to accommodate a ribbon-shaped serial ultrathin section (13).

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