Preparation method and device of double-arm clamping grid rack

Abstract

The present invention relates to a preparation method and device for a double-arm clamping grid carrier. The preparation method for the double-arm clamping grid carrier comprises the following steps: first, a hard material is made into a bracket with two adjacent bracket arms located in the same plane and in a clamping structure; then, the bracket is placed in a molten attachment liquid to obtain a bracket dipped in the attachment liquid; finally, the bracket dipped in the attachment liquid is retrieved, and the attachment liquid is solidified along the bracket arm to form an attachment layer. The device corresponding to the method of the present invention comprises a bracket and an attachment layer, and the attachment layer is attached to the inner side of the clamping structure portion between two adjacent bracket arms of the bracket.

Description

Technical Field

[0001] The invention relates to a grid rack for fixing a sample-carrying metal grid in a biological transmission electron microscope, and in particular to a device for fixing the sample-carrying metal grid with high throughput in a sample staining link. Background Art

[0002] Metal mesh is a tool for carrying samples in transmission electron microscopes, especially copper mesh, which is the most common. The copper mesh consists of two parts: one is a copper ring with an edge width of about 0.2mm, and the other is a hollow grid inside the copper ring. The entire copper mesh is about 18μm thick, and there is a layer of organic Fanghua membrane on the front of the copper mesh, which can effectively support the sliced ​​samples. Since the staining of biological samples needs to be carried out on the copper mesh, when staining, tweezers are usually used to clamp the 0.2mm ring width of the edge of the copper mesh. If the tweezers clamp the copper mesh grid area during this process, it is easy to break the Fanghua membrane attached to the grid surface, and then destroy the cell slices carried on the copper mesh, affecting the observation; and because the number of slices processed each time is large (4 to 20 slices, or even more), if each copper mesh is stained one by one, it will lead to the problem that the staining time of each slice is not easy to unify and the slices are easily mechanically damaged due to repeated operations.

[0003] In order to achieve uniform dyeing time for each slice and improve the problem of mechanical damage to the slices due to repeated operations, there is a device for batch dyeing of copper meshes in the prior art. Chinese invention patent specification CN105910875A discloses such a device for batch dyeing of copper meshes. The device includes a mesh carrier body and an auxiliary body, wherein the mesh carrier body is the part that fixes the sample-carrying copper mesh, and the mesh carrier auxiliary body is the part that fixes and supports the mesh carrier body. The mesh carrier body is composed of a hard layer and a soft layer. A soft layer is covered on the hard layer. The cross-section of the hard layer is arc-shaped or angle-shaped. The copper mesh is fixed in the device by using the soft layers distributed on both sides of the arc-shaped opening or the angle-shaped opening.

[0004] In the above device, the main body of the grid adopts a semi-enclosed structure, which makes the overall volume of the device larger, and ultimately requires more dye solution when using the device for batch dyeing; and except for the opening of the device, a groove or slit will be formed between the rest of the main body of the grid and the grid. When the device completes the dyeing operation and separates from the dye solution, the device will take out part of the dye solution, thereby further increasing the loss of the dye. In addition, the obstruction formed by the semi-enclosed structure of the grid frame will increase the length of the post-dyeing cleaning and air-drying process. Finally, the preparation of the above device is highly dependent on the mold; and because the soft layer is inside the semi-enclosed surrounding structure of the device, a complex process is required to achieve the adhesion and clamping efficiency of the soft layer. Summary of the invention

[0005] The main technical problem solved by the present invention is to provide a method and device for preparing a double-arm clamping grid rack which can reduce dye loss, improve the efficiency of cleaning and air-drying, simplify the device preparation process and realize batch dyeing of copper mesh.

[0006] In order to solve the above technical problems, the present invention provides a method for preparing a double-arm clamping grid frame, comprising the following steps:

[0007] a. A bracket made of a hard material having two adjacent bracket arms located in the same plane and having a clamping structure;

[0008] b. placing the stent into the molten attachment liquid to obtain a stent dipped in the attachment liquid;

[0009] c. Retrieve the stent dipped in the attachment liquid and wait for the attachment liquid to solidify into an attachment layer.

[0010] Furthermore, it also includes a method for reducing the loss of the adhesion layer and improving the efficiency of the device. Specifically, by removing the adhesion layer other than the inner sides of two adjacent bracket arms in the device, not only can the adhesion layer be recycled to the greatest extent, but the contact area between the copper mesh ring width and the wax layer can also be reduced, so that the pressure is greater and the copper mesh is easier to insert and remove.

[0011] Furthermore, the adhesion layer also includes a method for adjusting its thickness. Specifically, the thickness of the adhesion layer decreases with increasing temperature of the adhesion liquid, increases with increasing number of dipping times, increases with increasing dipping time, and increases with increasing degree of solidification of the wax liquid after each dipping except the last one.

[0012] A double-arm clamping grid-carrying frame device is composed of a support and an attachment layer. The attachment layer is attached to the inner side of a clamping structure portion between two adjacent support arms of the support.

[0013] Furthermore, the support is made of a hard material and provides a base surface for the attachment layer to attach and shape.

[0014] Furthermore, the attachment layer is composed of a soft material, and the soft attachment layer plays a role in fixing and supporting the copper mesh.

[0015] Furthermore, the cross-section of the device body is preferably in a single U shape.

[0016] Furthermore, the cross-section of the device body is preferably in a parallel spiral U-shape.

[0017] Furthermore, the interface of the device body is preferably in a closed ring shape.

[0018] In summary, the present invention has the following beneficial effects:

[0019] The present invention reduces the occupied volume of the device when in use by eliminating the redundant frame structure of the device, thereby reducing the amount of dye required for dyeing. At the same time, the open structure adopted by the present invention effectively avoids the formation of grooves or slits between the device and the copper mesh, thereby making the amount of dye carried out with the device close to zero. In addition, by adopting an open structure, the efficiency of the post-dyeing cleaning and air-drying steps is also improved. Furthermore, the preparation process of the device involved in the present invention is simpler, and almost zero dependence on the mold can be achieved when preparing the bracket; the clamping efficiency enhancement of the attachment layer can be quickly achieved by convenient processes such as directly removing the peripheral redundant wax layer along the bracket wall. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] The accompanying drawings are used to provide a further understanding of the present invention and constitute a part of the specification. Together with the following specific embodiments, they are used to explain the present invention but do not constitute a limitation of the present invention. In the accompanying drawings:

[0021] Figure 1 It is a diagram of a single U-shaped grid frame device and a cross-section of the device before efficiency enhancement.

[0022] Figure 2 It is a diagram of a single U-shaped grid frame device after efficiency enhancement and a cross-sectional view of the device.

[0023] Figure 3 It is a three-dimensional schematic diagram of a single U-shaped grid rack device carrying a copper grid.

[0024] Figure 4 It is a three-dimensional schematic diagram of a parallel spiral U-shaped grid rack device carrying a copper grid.

[0025] Figure 5 It is a three-dimensional schematic diagram of a closed annular grid-carrying frame device carrying a copper mesh.

[0026] Figure 6 It is a three-dimensional schematic diagram of batch dyeing of copper mesh. DETAILED DESCRIPTION

[0027] The following is a further detailed description of the specific implementation of the present invention in conjunction with the accompanying drawings and examples. The following examples are used to illustrate the present invention but are not intended to limit the scope of the present invention.

[0028] Example 1

[0029] The present invention discloses a method for preparing a double-arm clamping grid frame, which specifically comprises the following steps:

[0030] 1. Prefabricated bracket

[0031] A hard material (such as iron wire, steel wire, glass, plastic, etc.) is wound into a bracket comprising at least two bracket arms, and the two adjacent bracket arms of the bracket are located in the same plane, in a clamping structure, with a spacing of 3-4.5mm. The overall shape of the bracket is a single U-shape, a parallel spiral U-shape or a closed ring, etc., to provide a basis for the attachment and forming of the wax layer.

[0032] 2. Attach wax layer

[0033] a. Place solid wax (food grade beeswax or tooth wax, etc.) into a container and heat it;

[0034] b. After the solid wax is heated to a completely melted state, place the bracket into the wax liquid and dip it into the wax liquid;

[0035] Furthermore, the thickness of the stent wax layer can be adjusted by controlling the time, number of times the stent is dipped into the wax liquid, the temperature of the wax liquid, and the solidification degree of the wax liquid after each dipping except the last dipping.

[0036] Furthermore, as shown in Table 1, under the condition that the single dipping time, the number of dipping times and the degree of solidification of the wax liquid after each dipping except the last one are the same, the thickness of the wax layer is inversely proportional to the temperature of the wax liquid; under the condition that the wax liquid temperature, the single dipping time and the degree of solidification of the wax liquid after each dipping except the last one are the same, the thickness of the wax layer is proportional to the number of dippings; under the condition that the wax liquid temperature, the single dipping time and the degree of solidification of the wax liquid after each dipping except the last one are the same, the thickness of the wax layer is proportional to the number of dippings; under the condition that the wax liquid temperature, the single dipping time and the number of dippings are the same, the thickness of the wax layer is proportional to the degree of solidification of the wax liquid after each dipping except the last one.

[0037] Table 1 Wax layer thickness under different conditions

[0038]

[0039] d. Take the stent out of the wax solution and wait for the wax solution attached to the stent to solidify naturally;

[0040] 3. Reduce consumption and increase efficiency

[0041] like Figure 1 and Figure 2 As shown, taking the horizontal plane presented by the bracket as a reference, retain the wax layer attached to the inner side of the clamping structure between two adjacent bracket arms in the bracket, and cut off the wax layer attached to the front and back sides of the bracket, thereby reducing the thickness of the wax layer in contact with the copper mesh, reducing the contact area when the copper mesh is inserted into the wax layer, and increasing the pressure at the contact point between the mesh and the wax layer, which facilitates the insertion and removal of the copper mesh and can reduce the deformation of the copper mesh; cut off the wax layer attached to the outer edges of the two bracket arms of the bracket to recycle the wax and reduce the consumption of wax.

[0042] The ideal working thickness of the wax layer on the inner side between two adjacent bracket arms of the device is 0.5-1mm, and the ideal gap between the wax layers is 1-2.6mm. When the wax layer adsorbed by the entire bracket is too thick or the wax layer attached to the bottom of the bracket is too thick (after the bracket is taken out of the wax liquid, the wax liquid attached to it will flow downward under the action of gravity, causing the wax layer attached to the bracket to be thin on the top and thick on the bottom), the excessively thick part of the wax layer attached between two adjacent bracket arms of the device can be cut off according to actual needs to improve the efficacy or utilization rate of the device.

[0043] Example 2

[0044] Figure 3 Schematic diagram of a single U-shaped grid rack device carrying a copper mesh prepared according to the method of the present invention, the device consists of a single U-shaped bracket 1 and an attachment layer 2, the two bracket arms included in the bracket 1 provide a base surface for attachment and forming of the attachment layer 2, the attachment layer 2 provides effective fixing and support for the copper mesh 3, the copper ring part of the copper mesh 3 can be inserted and fixed on the inner side of the attachment layer 2, and arranged in sequence along the long axis of the bracket arm. Specifically, the single U-shaped bracket 1 is composed of hard materials (such as iron wire, steel wire, glass, plastic, etc.), the attachment layer 2 is composed of a wax layer (food-grade beeswax or tooth wax, etc.) and is attached to the inner side of the U-shaped structure of the single U-shaped bracket 1, the inner side spacing between the two bracket arms of the single U-shaped bracket 1 is 3 to 4.5 mm, the inner side spacing between the attachment layer 2 is 1 to 2.6 mm, and the edge of the copper mesh 3 is a copper ring with an outer diameter of 3 mm, an inner diameter of 2.6 mm, and a ring width of about 0.2 mm; there are hollow grids with mesh sizes ranging from 50 to 400 within the ring width; the entire copper mesh 3 is about 18 μm thick; there is a layer of organic flannel film on the front of the copper mesh 3, which can effectively support the sliced ​​sample.

[0045] Figure 4 Schematic diagram of a parallel spiral U-shaped grid rack device carrying a copper mesh prepared according to the method of the present invention, the device consists of a parallel spiral U-shaped bracket 1 and an attachment layer 2, the two adjacent bracket arms in the bracket 1 provide a base surface for attachment and forming of the attachment layer 2, the attachment layer 2 provides effective fixing and supporting effects on the copper mesh 3, the copper ring part of the copper mesh 3 can be inserted and fixed on the inner side of the attachment layer 2, and arranged in sequence along the long axis of the bracket arm. Specifically, the parallel spiral U-shaped bracket 1 is composed of hard materials (such as iron wire, steel wire, glass, plastic, etc.), the attachment layer 2 is composed of a wax layer (food-grade beeswax or tooth wax, etc.) and is attached to the inner side of the U-shaped structure of the parallel spiral U-shaped bracket 1, the inner side spacing between two adjacent bracket arms of the parallel spiral U-shaped bracket 1 is 3 to 4.5 mm, the inner side spacing of the attachment layer 2 is 1 to 2.6 mm, and the edge of the copper mesh 3 is a copper ring with an outer diameter of 3 mm, an inner diameter of 2.6 mm, and a ring width of about 0.2 mm; there are hollow grids with mesh sizes ranging from 50 to 400 within the ring width; the entire copper mesh 3 is about 18 μm thick; there is a layer of organic film on the front of the copper mesh 3, which can effectively support the sliced ​​sample.

[0046] Figure 5 Schematic diagram of a closed annular grid-carrying rack device carrying a copper mesh prepared according to the method of the present invention, the device consists of a closed annular bracket 1 and an attachment layer 2, the two bracket arms of the bracket 1 provide an attachment base surface for the attachment layer 2, the attachment layer 2 provides effective fixing and supporting effects on the copper mesh 3, the copper ring part of the copper mesh 3 can be inserted and fixed on the inner side of the attachment layer 2, and arranged in sequence along the long axis of the bracket arm. Specifically, the closed ring-shaped bracket 1 is composed of hard materials (such as iron wire, steel wire, glass, plastic, etc.), the adhesion layer 2 is composed of a wax layer (food-grade beeswax or tooth wax, etc.) and is attached to the inner sides of the two bracket arms of the closed ring-shaped bracket 1, the inner side spacing of the two bracket arms of the closed ring-shaped bracket 1 is 3 to 4.5 mm, the inner side spacing of the adhesion layer 2 is 1 to 2.6 mm, and the edge of the copper mesh 3 is a copper ring with an outer diameter of 3 mm, an inner diameter of 2.6 mm, and a ring width of about 0.2 mm; there are hollow grids with mesh sizes ranging from 50 to 400 within the ring width; the entire copper mesh 3 is about 18 μm thick; there is a layer of organic flannel film on the front of the copper mesh 3, which can effectively support the sliced ​​sample.

[0047] Figure 6 It is a schematic diagram of batch dyeing of copper meshes according to the present invention, wherein the spacing between adjacent copper meshes fixed on the attachment layer is greater than 2.5 mm to reduce the carry-out of dye solution 4; the portion of the device carrying the copper mesh 3 is immersed below the liquid surface of the dye solution 4 to achieve batch dyeing of the copper meshes.

Claims

1. A method for preparing a double-arm clamping grid frame, characterized in that: The following steps are involved: a. A hard material is made into a bracket (1), wherein the bracket comprises at least two bracket arms, and two adjacent bracket arms are located in the same plane and are in a clamping structure, with a spacing of 3-4.5 mm, and the diameter of the bracket arm is smaller than the outer diameter of the copper mesh; b. placing the stent into the molten attachment liquid to obtain a stent dipped in the attachment liquid (1); c. Retract the support (1) dipped in the attachment liquid, and wait for the attachment liquid to solidify into an attachment layer (2), wherein the attachment layer (2) is used to fix and support the copper mesh.

2. The method according to claim 1, characterized in that Step c also includes removing the attachment layer (2) attached to the support (1) other than the inner sides of two adjacent support arms.

3. A double-arm clamping grid-carrying frame device, comprising a support (1) and an attachment layer (2), characterized in that: The support (1) is composed of at least two support arms and a connecting portion, the two adjacent support arms on both sides of the connecting portion are located in the same plane and are in a clamping structure, with a spacing of 3-4.5 mm, the diameter of the support arm is smaller than the outer diameter of the copper mesh, the attachment layer (2) is attached to the inner side of the clamping structure between the two adjacent support arms of the support, and the attachment layer (2) is used to fix and support the copper mesh.

4. The device according to claim 3, characterized in that The support (1) is made of a hard material.

5. The device according to claim 3, characterized in that The adhesion layer (2) is composed of a soft material.

6. The device according to claim 3, characterized in that The longitudinal section of the device is in a single U shape.

7. The device according to claim 3, characterized in that The longitudinal section of the device is in a parallel spiral U shape.

8. The device according to claim 3, characterized in that The longitudinal section of the device is in the form of a closed ring.

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