JP2001148228A - Device for putting on grid ultra-thinly sliced piece of specimen for electron microscope - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a device which can be used for a grid to which a supporting membrane is spread while this could not be used in the previous loop method and which can be also used for a mono-hole grid to which a supporting membrane is spread, and which can be rapidly dried where the water between a ring and the grid is almost absorbed by a blotting paper. SOLUTION: A circular hole 1 is opened that has the same diameter as a metallic grid in which the outside diameter is used, and in which the thickness is 20 μm-50 μm, and that has the range to enable the observation with an electron microscope in the center. A support axis 3 is equipped at one side of the ring 2 which is formed to have a very low slope toward outside, and a grip of pencil-like thickness is installed. The angle made by the boundary part between the ring and the support axis is decided as one's wish so that the border of the ring becomes horizontal, when the one holds the handle of the device for using it. In the case of the grid to which the supporting membrane is spread, the diameter of the circular hole which is opened in the center is made to have the same diameter as the hole diameter of the used mono-hole grid.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a tool for ultra-slicing a sample for observation with an electron microscope by an ultra-thin section method, and placing an ultra-thin section floating on the water surface of a knife boat on a grid.

[0002]

2. Description of the Related Art To observe a tissue with an electron microscope, an ultrathin section prepared by an ultrathin section method is used. In the ultra-thin section method, a tissue is cut into 1 mm ³ , and a section sliced to a thickness of about 0.1 μm or less is observed by electronic staining. Briefly, a series of steps up to electron microscopic observation is fixed (osmic acid, glutaraldehyde) → water washing (buffer) → dehydration (alcohol) → embedding (epoxy resin, other resin)
→ Ultra-slicing (ultra-microtome) → load (with or without grid and supporting membrane) → electron staining (heavy metal) → observation (electron microscope) →
Photo shoot. When observing with an electron microscope, ultra-thin sections that are easily torn are placed on a grid with a support film placed on them and observed.However, the resulting images have poor resolution. Observe. Even if the ultrathin section is difficult to break, the ultrathin section that is not stretched over the entire surface of the grid hole is torn at the time of observation, or the ultrathin section moves due to shrinkage, so that a photograph cannot be taken. Good tensioning should cover the entire surface of the grid holes. The following method is used to stretch an ultrathin section into a hole in a grid. In the pressing method, the edge of the grid that has been subjected to the adhesive treatment is pinched with tweezers, and the adhesive surface is pressed against an ultrathin section floating on the water surface. This method moves the ultra-thin section instantaneously by contacting the water before contacting the ultra-thin section in order to bring the grid close to the water surface where the ultra-thin section floats exactly in parallel. It is stretched out of the hole of the target grid. In addition, since the water surface is technical at the time of ultra-slicing, the concave surface for lowering the water surface of the knife boat from the cutting edge promotes that the water surface is not stretched as intended. In the scooping method, grip the edge of the previously hydrophilic grid with tweezers, submerge the grid near the ultra-thin section floating on the water to the position where it is stuck to the surface, make it vertical, and use tools such as eyelashes on the grid surface The method of attaching the edge of an ultra-thin section and pulling it vertically, also known as the pulling method. It is suitable for stretching one wide ultra-thin section on a grid with small holes, but not suitable for stretching several small ultra-thin sections on a grid with small holes. The loop method uses a tool having a flat loop with a handle made of an extremely thin plate having an outer diameter of 3 mm, which is the same as that of the grid, and an inner diameter of 2 mm, which can be observed by an electron microscope. An ultra-thin section floating on the water surface is placed in the inner diameter of the ring, landed on water, and pulled up. When the lower part of the water droplet of the ring is gently brought into contact with the prepared grid, the grid adheres concentrically via the water droplet. The blotting paper is pressed against the side of the ring to absorb water droplets from the gap with the grid. Alternatively, when the grid is pressed against the blotting paper with a ring, water droplets are absorbed by the blotting paper from the holes in the grid. If the grid is not covered with a support membrane, it will fall under the weight of the grid when it dries. In a grid with a supporting film, the ring is a flat surface, so that the supporting film does not stick to the ring side and does not fall due to the weight of the grid. When this is forcibly separated, the support film stretched over the grid holes adheres to the ring. Even in the region where the ultra-thin section inside the ring is placed, if the inner edge of the ring comes into contact with the support film and is partially broken by the grid hole, the electron beam will cause the support film to shrink during observation and the tear will spread. Therefore, it is not used for ultra-thin sections requiring a support film because the sample cannot be taken due to the movement of the sample. Further, there is a limit that water between the ring and the grid can be absorbed by the blotting paper. For this purpose, holes are provided on the loop surface to take measures against drying, but there is a disadvantage that drying takes time.

[0003]

The present invention can be used for a grid with a support membrane and a single-hole grid with a support membrane which could not be used in the conventional loop method, and absorbs water between the ring and the grid. It is intended to provide a tool which is almost completely absorbed by paper and dries quickly.

[0004]

The present invention will be described with reference to the drawings. A) The same diameter as that of a grid (7) made of a metal having a thickness of 20 μm to 50 μm and having an outer diameter, and within a range that can be observed by an electron microscope in the center. A circular hole (1) is drilled, and a support shaft (3) is provided on one side of a ring (2) formed with a very gentle slope lowered outward, and a thick handle (4) is provided on the support shaft (3). Is provided. When holding the tool handle (4) in hand, use the border of the wheel (2) and the spindle (3) to your liking so that the edge of the wheel (2) is almost horizontal. Make an angle with. B) In the case of a single-hole grid with a support membrane, the diameter of a circular hole (1) drilled in the center should be the same as the diameter of the single-hole grid to be used. The present invention is a tool for mounting an ultra-thin section of an electron microscope sample having the above configuration on a grid.

[0005]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Since the present invention has the above-mentioned structure, when using the same, the handle (4) of the present invention is gripped and the ring (2) is placed inside the ring (2).
Ultra-thin section (6) floating on the water surface (5) of a knife boat
And landed on it and lifted it (see FIGS. 5 and 6). When the lower part of the water droplets of the ring is gently brought into contact with the grid which has been subjected to the adhesive treatment or the grid (7) on which the support film is stretched, the grid (7) adheres concentrically via the water droplets (see FIG. 7). The loop (2) is inclined to bring the blotter (8) into contact with the lower side to absorb water droplets from the gap between the loop (2) and the grid (7) (see FIGS. 8 and 9). When absorbing the water droplets, the ultrathin section (6) is attached to the grid surface in the area inside the loop (2) with water. Since the wheel (2) has a low and gentle slope toward the outside, the outer periphery of the grid (7) comes into contact with the wheel (2).
Since the gap between the grid and the center is wide near the center of the width, the water flows downward and most of the water is absorbed by the blotting paper (8), so that the water dries quickly (see FIG. 10). In order to prevent the grid (7) from dropping from the wheel (2), the grid (7) is moved up to the wheel (2) to the sample storage grid case (see FIG. 11).

[0006]

As described above, since the ring has a gentle slope, if water is absorbed by blotting paper, the edge of the grid and the edge of the ring come into contact with each other. Even in the case of a single-hole grid with a membrane, the surface of the ring and the surface of the support film do not adhere to each other. Since the physical property that water between the grid and the ring moves toward the narrow gap and the gap where the water flows between the grid and the ring are secured, the water is efficiently absorbed by the blotting paper. The present invention is an effect due to the gentle inclination of the ring.

[Brief description of the drawings]

FIG. 1 is a plan view of the present invention.

FIG. 2 is a front view of the present invention.

FIG. 3 is an enlarged perspective view of a ring portion.

FIG. 4 is an enlarged perspective view of a partial cross section of a ring.

FIG. 5 is an enlarged perspective view of an ultra-thin section of the water surface of a knife boat stored in a wheel.

FIG. 6 is an enlarged side view immediately before contact with a grid.

FIG. 7 is an enlarged side view after touching a grid.

FIG. 8 is an enlarged side view showing a state in which water is absorbed by a blotter through a gap between a grid and a ring.

FIG. 9 is an enlarged sectional view showing a state in which water is absorbed by a blotter through a gap between a grid and a ring.

FIG. 10 is an enlarged cross-sectional view before water is completely absorbed by a blotter through a gap between a grid and a ring.

FIG. 11 is an enlarged cross-sectional view in which a grid is placed on a ring for movement after absorbing water with blotting paper.

FIG. 12 is a front view in which an ultrathin section is adhered to a grid on which a support film is not provided.

FIG. 13 is an enlarged sectional view showing a state before water is completely absorbed by blotting paper from a gap between a single-hole grid and a ring.

FIG. 14 is a front view of an ultrathin section mounted on a support membrane stretched over a single-hole grid.

[Explanation of symbols]

 (1) Hole (2) Ring (3) Spindle (4) Handle (5) Knife boat water surface (6) Ultra-thin section (7) Grid (8) Blotting paper (9) Water (10) Support membrane

Claims (1)

[Claims] An extremely thin circular plate is provided with a circular hole (1) in the center, and a support shaft (3) is provided on one side of a ring (2) formed with a very gentle inclination toward the outside. The handle (4) is provided on the support shaft (3). A tool configured as described above for mounting an ultra-thin section of an electron microscope sample on a grid.