Micro-reaction chamber, multi-group parallel micro-reaction chamber and sample observation method

Abstract

The invention relates to the technical field of micro-reaction cavities, in particular to a micro-reaction cavity, multiple groups of parallel micro-reaction cavities and a sample observation method.The micro-reaction cavity comprises a shell, an upper partition piece and a lower partition piece are arranged in the shell, and an inner cavity of the shell is divided into an upper cavity located at the top, a reaction cavity located in the middle and a lower cavity located at the bottom; the distance between the upper separator and the lower separator can be adjusted, so that the bending conditions of the upper separator and the lower separator of the reaction cavity in which the sample is positioned can be independently changed by adjusting the pressure values of the upper chamber and the lower chamber under the condition of ensuring the set pressure of the reaction cavity in which the sample is positioned, and the bending conditions of the upper separator and the lower separator of the reaction cavity in which the sample is positioned can be independently changed under the condition of changing the sample pressure. The lower separator attached to the sample is maintained not to generate vertical displacement, so that under the condition that the sample is not out of focus, the situations of electron scattering and absorption can be reduced at the same time, and the electronic imaging quality and imaging stability are improved.

Description

Technical Field

[0001] The present disclosure relates to the technical field of micro-reaction chambers, and in particular to a micro-reaction chamber, a plurality of groups of micro-reaction chambers connected in parallel, and a sample observation method. Background Art

[0002] Electron microscope, referred to as EM, has significantly higher magnification and resolution than optical microscope, so it is widely used in disease diagnosis, material analysis, substance identification and other fields. According to the structure and use, electron microscopes can be divided into scanning electron microscope (SEM), transmission electron microscope (TEM), scanning transmission electron microscope (STEM), etc. Among them, TEM has the highest magnification and resolution, and can be used to observe biochemical and material reaction processes at the atomic scale. However, biochemical and material reactions mostly occur in gas or liquid environments, such as the observation of viruses and proteins in liquids, the reaction between electrode materials and electrolytes in lithium batteries, and the role of catalysts in gas reactions.

[0003] In order to solve the above problems and make in-situ observation possible, people usually use in-situ sample observation methods based on double-layer membrane structure micro-reactors. At present, the micro-reactor solution providers for electron microscopes mainly include DENS-solution in the Netherlands and Protochips in the United States. Figure 1 As shown. The principle is to confine the reaction material in a sealed thin layer space, and set an electron-permeable thin film window on the surface of the reaction chamber, allowing the electron beam to pass through the window to observe the sample inside the reaction chamber. This method can image and observe solid, gaseous, and liquid substances inside the reaction chamber while ensuring the vacuum environment inside the electron microscope. Further systems will also increase the temperature and magnetic field control inside the cavity, and use pipes to connect the reaction chamber to the gas and liquid supply system, so that the ambient temperature control, material exchange, and pressure control inside the reaction chamber can be achieved, thereby dynamically simulating the actual reaction process. This is what we usually call dynamic in-situ observation.

[0004] A core idea of ​​in-situ electron microscopy or X-Ray microscopy is to ensure that the sample is in the environment set for the experiment, in which pressure P is an important environmental variable. That is, the thickness of the reaction chamber needs to be adjusted under the premise that the sample pressure (gas or liquid) is the set value. This is especially important for pressure-sensitive samples. For example, in catalyst experiments, gas pressure is an important control factor for catalyst efficiency and working status. For example, in living cell observations, appropriate liquid pressure is a necessary factor for cell survival and life behavior. It is unacceptable to change the pressure of the reaction chamber at will.

[0005] In the existing reaction chamber based on the double-layer membrane structure, since the outside of the reaction chamber is a vacuum and the inside of the reaction chamber is a specified pressure, the double-layer membrane structure of the reaction chamber will bend toward the outside of the reaction chamber, and the degree of bending increases as the pressure in the reaction chamber increases. Since the sample is placed on the film, when the pressure in the reaction chamber is adjusted, the sample will have a vertical displacement and needs to be refocused, making it difficult to image continuously. Pressure and bending are causally related, so they cannot be controlled independently, that is, it is impossible to control the bending of the membrane while maintaining the pressure in the reaction chamber at a set value, and the corresponding thickness of the reaction chamber cannot be controlled. Since the thickness of the reaction chamber cannot be controlled, it is difficult to effectively thin the liquid layer when the inside of the reaction chamber is liquid. Due to the heat dissipation and absorption of the electron beam by the liquid, imaging is impossible or the imaging quality is poor.

[0006] The prior art also provides a reaction chamber based on a three-layer membrane structure, in which a support structure specifically used to carry the sample is inserted into the original cavity. Since the pressure on the upper and lower sides of the sample support structure is always the same, it will not move vertically with the change of the reaction chamber pressure. However, there is still no reaction chamber structure that can control the sample thickness while ensuring that the focus does not shift. Summary of the invention

[0007] In order to solve the problems in the related art, the embodiments of the present disclosure provide a micro-reaction chamber, a plurality of groups of micro-reaction chambers connected in parallel, and a sample observation method.

[0008] In a first aspect, a micro-reaction chamber is provided in an embodiment of the present disclosure, comprising:

[0009] The shell is provided with an upper separator and a lower separator, which divide the inner cavity of the shell into an upper chamber located at the top, a reaction chamber located in the middle and a lower chamber located at the bottom, and the distance between the upper separator and the lower separator can be adjusted.

[0010] Optionally, the upper chamber, the reaction chamber, and the lower chamber are all gas chambers; or, the upper chamber and the lower chamber are gas chambers, and the reaction chamber is a liquid chamber.

[0011] Optionally, the upper separator and / or the lower separator is at least partially a membrane structure, and the membrane structure has a thinned area to form an electron beam observation area.

[0012] Optionally, a portion of the upper surface and / or lower surface of the upper separator is thinned in a thickness direction to form the thinned region; and / or,

[0013] A partial area of ​​the upper surface and / or the lower surface of the lower separator is thinned in a thickness direction to form the thinned area.

[0014] Optionally, the upper separator and / or the lower separator is composed of an upper layer and a support plate, the upper layer is a membrane structure, the upper layer is thinned along the thickness direction and forms the thinned area with the support plate, or the support plate is thinned along the thickness direction and forms the thinned area with the upper layer, or the upper layer and the support plate are each thinned along the thickness direction to form the thinned area.

[0015] Optionally, the upper separator and / or the lower separator is / are a non-crystalline carbon film or an elastic polymer film.

[0016] Optionally, the thickness of the thinned region is 0.3-100 nm, and / or the thickness of the upper separator and / or the lower separator is less than 1 μm, preferably 20-100 nm.

[0017] Optionally, the thickness of the upper separator and / or the lower separator is smaller than the thickness of the shell.

[0018] Optionally, the shell has a thickness of 20-1000 nm, preferably 20-100 nm, and the upper separator and / or the lower separator has a thickness of 0.3-100 nm, preferably 5-20 nm.

[0019] Optionally, the shell is at least partially a membrane structure.

[0020] Optionally, the upper chamber, reaction chamber, and lower chamber are each connected to a pressure control device.

[0021] Optionally, the pressure value in the reaction chamber is a set value, the pressure value in the upper chamber is greater than, equal to or less than the set value, and the pressure value in the lower chamber is greater than, equal to or less than the set value.

[0022] Optionally, an observation window is provided on the shell to adapt to focused electron microscopy, optical microscopy or X-Ray microscopy for sample observation.

[0023] In a second aspect, a plurality of groups of micro-reaction chambers connected in parallel are provided in an embodiment of the present disclosure, comprising a plurality of micro-reaction chambers as described in any one of the first aspect.

[0024] In a third aspect, an embodiment of the present disclosure provides a method for adjusting a micro-reaction chamber, wherein the micro-reaction chamber is any one of the micro-reaction chambers described in the first aspect, comprising the following steps:

[0025] a step of regulating the pressure in the reaction chamber;

[0026] a step of regulating pressure in the upper chamber and / or the lower chamber;

[0027] Steps for sample observation.

[0028] Optionally, in the step of adjusting the pressure in the reaction chamber, the pressure value in the reaction chamber is adjusted to a predetermined pressure value;

[0029] In the step of adjusting the pressure in the upper chamber and / or the lower chamber, the pressure value in the lower chamber is adjusted to the predetermined pressure value, and the pressure value in the upper chamber is adjusted to a value greater than the predetermined pressure value;

[0030] In the step of observing the sample, the sample is observed under the conditions of the pressure value in the upper chamber, the pressure value in the reaction chamber, and the pressure value in the lower chamber.

[0031] According to the technical solution provided in the embodiment of the present disclosure, the distance between the upper partition and the lower partition can be adjusted. By ensuring that the reaction chamber where the sample is located is at a set pressure, the bending conditions of the upper partition and the lower partition of the reaction chamber where the sample is located can be independently changed by adjusting the pressure values ​​of the upper chamber and the lower chamber. It is also possible to maintain the lower partition to which the sample is attached without vertical displacement under the condition of changing the sample pressure. This can reduce electron scattering and absorption while keeping the sample in focus, thereby improving the quality and stability of electronic imaging.

[0032] It is to be understood that the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the present disclosure. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Other features, objectives and advantages of the present disclosure will become more apparent through the following detailed description of non-limiting embodiments in conjunction with the accompanying drawings. In the accompanying drawings:

[0034] Figure 1 The schematic diagram of the structure of the micro-reaction chamber in the prior art is shown.

[0035] Figure 2 A schematic structural diagram of a micro-reaction chamber according to an embodiment of the present disclosure is shown.

[0036] Figure 3 A schematic diagram showing a thinning area according to an embodiment of the present disclosure.

[0037] Figure 4 A schematic diagram showing a thinned region according to another embodiment of the present disclosure.

[0038] Figure 5 Show Figure 1 The assembly flow chart of the micro-reaction chamber is shown.

[0039] Figure 6 A schematic diagram showing the structure of multiple groups of parallel micro-reaction chambers according to an embodiment of the present disclosure.

[0040] Figure 7 A schematic diagram showing a first usage state of the micro-reaction chamber according to an embodiment of the present disclosure.

[0041] Figure 8 A schematic diagram showing a second use state of the micro-reaction chamber according to an embodiment of the present disclosure.

[0042] Fig. 9 A schematic diagram showing a third usage state of the micro-reaction chamber according to an embodiment of the present disclosure.

[0043] Fig.10 A schematic diagram showing a fourth use state of the micro-reaction chamber according to an embodiment of the present disclosure.

[0044] Fig.11 A schematic diagram showing a fifth use state of the micro-reaction chamber according to an embodiment of the present disclosure. DETAILED DESCRIPTION

[0045] Hereinafter, exemplary embodiments of the present disclosure will be described in detail with reference to the accompanying drawings so that those skilled in the art can easily implement them. In addition, for the sake of clarity, parts not related to the description of the exemplary embodiments are omitted in the accompanying drawings.

[0046] In the present disclosure, it should be understood that terms such as "including" or "having" are intended to indicate the presence of features, numbers, steps, behaviors, components, parts, or combinations thereof disclosed in the present specification, and are not intended to exclude the possibility that one or more other features, numbers, steps, behaviors, components, parts, or combinations thereof exist or are added. It should also be noted that the embodiments in the present disclosure and the features therein may be combined with each other in the absence of conflict.

[0047] The present disclosure will be described in detail below with reference to the accompanying drawings and in conjunction with embodiments.

[0048] Figure 2 A schematic structural diagram of a micro-reaction chamber according to an embodiment of the present disclosure is shown.

[0049] like Figure 2The micro-reaction chamber 1 comprises: a shell 2, in which an upper partition 3 and a lower partition 4 are arranged, dividing the inner cavity of the shell into an upper chamber 5 located at the top, a reaction chamber 6 located in the middle, and a lower chamber 7 located at the bottom, wherein the upper chamber 5, the reaction chamber 6, and the lower chamber 7 contain pressure-controllable fluids, such as liquids or gases, wherein the pressure value in the upper chamber 5 is P1, the pressure value in the reaction chamber 6 is P2, and the pressure value in the lower chamber 7 is P3, and by changing the pressure difference among the pressure values ​​P1, P2, and P3, the distance between the upper partition 3 and the lower partition 4 (or the thickness of the reaction chamber) can be adjusted, for example, the thickness of the reaction chamber can be increased so that the sample in the reaction chamber can react fully, while reducing the thickness of the reaction chamber can reduce the influence of the fluid in the reaction chamber on the imaging quality, thereby obtaining a high-resolution image. Since three chambers are formed, the pressure value P2 in the reaction chamber 6 can be set as needed, and the thickness of the reaction chamber can be adjusted by adjusting the pressure difference among the pressure values ​​P1, P2, and P3, thereby achieving independent adjustment of the sample pressure and the thickness of the reaction chamber.

[0050] In the disclosed method, the upper chamber 5, the reaction chamber 6, and the lower chamber 7 can all be gas chambers; or, the upper chamber 5 and the lower chamber 7 are gas chambers, and the reaction chamber 6 is a liquid chamber. The reaction chamber 6 is a gas chamber or a liquid chamber to adapt to the observation of samples in different biochemical and material reactions, such as the observation of viruses and proteins in liquids, the reaction of electrode materials and electrolytes in lithium batteries, the role of catalysts in gas reactions, etc.

[0051] In the disclosed method, the upper separator 3 and / or the lower separator 4 are membrane structures, that is, they are deformed under pressure difference to achieve the change of the thickness of the reaction chamber. Further, the upper separator 3 and / or the lower separator 4 can be a thin film material as a whole, such as an amorphous carbon film or an elastic polymer film, with a thickness of less than 1 micron, preferably 20-100nm, or a part of the area is a thin film material. In this case, the sample only needs to be placed in the membrane structure area, and when imaging the sample, the thickness of the reaction chamber can be reduced to obtain a high-resolution image.

[0052] Furthermore, the film structure of the upper separator 3 and / or the lower separator 4 has a thinned region to form an electron beam observation region, and the thickness of the thinned region is 0.3-100 nm.

[0053] Figure 3 A schematic diagram showing a thinning area according to an embodiment of the present disclosure. Figure 3 The above separator 3 is used as an example for schematic illustration. It is understandable that the lower separator 4 may also adopt the same structure, and the present disclosure does not limit this. Figure 3As shown, a part of the upper surface of the upper separator 3 is thinned in the thickness direction to form a thinning area a, and the sample is placed in the reaction chamber 6 opposite to the thinning area a. When observing the sample, the electron beam is focused from the thinning area a to the sample, so as to improve the observation accuracy. Similarly, a part of the lower surface of the upper separator 3 can also be thinned in the thickness direction to form a thinning area b, or the corresponding areas of the upper and lower surfaces of the upper separator 3 are simultaneously thinned in the thickness direction to form a thinning area (not shown in the figure), so as to improve the observation accuracy. It should be noted that the range and number of the thinning areas are preferably set so as not to affect the overall strength of the upper separator 3 and the lower separator 4 (to adapt to the subsequent pressure adjustment), and can be flexibly adjusted as needed. By setting a thinning area on the upper separator 3 and / or the lower separator 4, the sample is placed in the thinning area, so that the electrons pass through the thinning area to the sample for imaging, which is conducive to improving the imaging quality and facilitating sample observation.

[0054] Figure 4 A schematic diagram showing a thinned region according to another embodiment of the present disclosure. Figure 4 The following separator 4 is used as an example for schematic illustration. It is understood that the upper separator 3 may also adopt the same structure, and the present disclosure does not limit this. Figure 4 As shown, the lower separator 4 is composed of an upper layer 41 of a membrane structure and a support plate 42 that plays a supporting role. The present disclosure only uses the upper layer 41 and the support plate 42 as examples for schematic illustration. Of course, the support plate 42 can also be located above the upper layer 41. It can be understood that the lower separator 4 can also be a single layer or other multi-layer structure, which will not be described in detail here. Among them, the upper layer 41 can be thinned in the thickness direction and form a thinning area c with the support plate 42 below. Similarly, the support plate 42 can be thinned in the thickness direction and form a thinning area d with the upper layer 41 above. In some cases, the upper layer 41 and the support plate 42 can be thinned in the thickness direction at the same time to form a thinning area (not shown in the figure). The range and number of the thinning area are set so as not to affect the overall strength of the upper separator 3 and the lower separator 4 (to adapt to the subsequent pressure adjustment), and can be flexibly adjusted as needed. The present disclosure will not be described in detail here. The combination of the support plate and the film layer makes the process simpler and it is also easier to obtain a thinning area of ​​the required thickness. Moreover, different materials can be selected for the support plate and the upper layer, so that they can withstand the pressure difference and produce corresponding deformation in the subsequent pressure adjustment process, and the thickness of the thinned area can be thin enough to improve the imaging quality.

[0055] In the disclosed method, the material of the membrane structure can be selected from one or more of silicon nitride, silicon carbide, aluminum oxide, carbon film, graphene, etc., and the thickness of the membrane structure ranges from 0.3nm to 1μm. The material of the support plate can also be selected from one or more of silicon nitride, silicon carbide, aluminum oxide, carbon film, graphene, etc., with a thickness range of 50nm to 1μm. The thinning area is set to allow the electron beam to be transmitted, and the thickness of the thinning area is 0.3nm-100nm. Among them, the 0.3nm material can be selected from a single layer of graphene.

[0056] For example, the support plate 42 is made of 200-400 nm silicon nitride, and the thinned region (upper layer 41) is made of 10-20 nm silicon nitride or 5-10 nm aluminum oxide or single-layer / multi-layer graphene material.

[0057] In the disclosed method, the shell 2 is at least partially a membrane structure. The upper separator 3 and / or the lower separator 4 are at least partially a membrane structure. Specifically, the shell 2 can be formed by sealing the upper shell 21 and the lower shell 22, wherein at least a part of the upper shell 21 or the lower shell 22 is a thin film material, and similarly, at least a part of the upper separator 3 and / or the lower separator 4 is a thin film material, which is convenient for adapting to focusing electron microscopy, optical microscope or X-Ray microscope for sample observation.

[0058] In the disclosed method, the thickness of the upper separator 3 and / or the lower separator 4 is less than the thickness of the shell 2, that is, the upper shell 21 and the lower shell 22 can choose a relatively thicker pressure-resistant film. Since they are far away from the sample and are not in the focusing plane, the impact on the imaging quality can be optimized by adjusting the imaging equipment and parameters, and the upper separator 3 and / or the lower separator 4 selects a thinner material that is more suitable for imaging, which is beneficial to improving the imaging quality. For example, the shell thickness can be 20-1000nm, preferably 20-100nm, and the thickness of the upper separator 3 and / or the lower separator 4 is 0.3-100nm, preferably 5-20nm. It should be noted that the shell can also be thinned in the thickness direction to form a thinned area. At this time, the thickness of the thinned shell is between 20-100nm, and the thickness of the remaining part of the shell can be between 100nm-1000nm. The thinned area of ​​the shell needs to correspond to the thinned area of ​​the upper separator 3 and / or the lower separator 4 to ensure that the electron beam irradiates the sample from the thinned area for imaging, thereby obtaining a higher imaging quality.

[0059] In the following, unless otherwise specified, the three chambers formed by the four-layer membrane structure composed of the shell 2, the upper separator 3 and the lower separator 4, namely the upper chamber 5, the reaction chamber 6 and the lower chamber 7 are schematically described as an example.

[0060] A four-layer membrane structure is adopted to form three chambers. The upper chamber 5, the reaction chamber 6, and the lower chamber 7 are respectively connected to a pressure control device to independently adjust the pressure in the chamber, wherein the pressure value P2 in the reaction chamber 6 is a set value, the pressure value in the upper chamber 5 can be greater than, equal to, or less than the set value, and the pressure value in the lower chamber 7 can be greater than, equal to, or less than the set value.

[0061] like Figure 7 As shown, the pressure value P1=P2=P3 is adjusted. When the sample is observed, the micro-reaction chamber is in a vacuum environment, the upper shell 21 and the lower shell 22 will expand outward, and the partition 3 and the lower partition 4 are not bent. This is considered to be the initial thickness of the reaction chamber. As an example, those skilled in the art can know that the initial thickness of the reaction chamber can also be adjusted when the external environment of the micro-reaction chamber is a non-vacuum environment, such as under normal pressure, and the focal position can be guaranteed not to change.

[0062] like Figure 8 As shown, by adjusting the pressure value P1<P2=P3, since P2=P3, the lower partition 4 where the sample is located can be maintained without vertical displacement, ensuring that the sample is not out of focus. By adjusting the value P1<P2, the upper partition 3 bends outward, and the thickness of the reaction chamber increases compared to the initial thickness. The fluid capacity in the reaction chamber also increases, which facilitates the sample to fully react.

[0063] like Fig. 9 As shown, by adjusting the pressure value P1>P2=P3, since P2=P3, the lower partition 4 where the sample is located can be maintained without vertical displacement, ensuring that the sample is not out of focus. By adjusting the value P1>P2, the upper partition 3 bends inward, and the thickness of the reaction chamber is reduced compared to the initial thickness, and the thickness of the fluid in the reaction chamber is thinned accordingly. While ensuring that the sample is not out of focus, the influence of the fluid in the reaction chamber on the imaging quality can be reduced, thereby obtaining a high-resolution image.

[0064] like Fig.10 As shown, the pressure values ​​are adjusted to P1<P2, P3<P2, the upper partition 3 and the lower partition 4 are bent outward, the thickness of the reaction chamber is increased compared to the initial thickness, and the fluid capacity in the reaction chamber is increased accordingly, which is convenient for the sample to fully react.

[0065] like Fig.11 As shown, by adjusting the pressure values ​​P1>P2 and P3>P2, the upper partition 3 and the lower partition 4 bend inward, the thickness of the reaction chamber is reduced compared to the initial thickness, and the thickness of the fluid in the reaction chamber is thinned accordingly, so as to reduce the influence of the fluid in the reaction chamber on the imaging quality, thereby obtaining a high-resolution image.

[0066] It should be noted that Fig.10 , Fig.11In some scenarios, the sample may be out of focus. In this case, you only need to refocus the sample and then perform imaging to obtain a high-resolution image.

[0067] In the disclosed method, an observation window (not shown in the figure) is opened on the shell 2 to adapt to focused electron microscopy technology, optical microscope or X-Ray microscope for sample observation.

[0068] In the disclosed method, at least one through hole may be provided on the lower partition 4 to connect the reaction chamber and the lower chamber, so that the pressure value P2=P3. The reaction chamber and the lower chamber do not need to be connected to pressure control devices respectively, which reduces one pressure control device. The pressure value can be adjusted synchronously using one pressure control device, simplifying the structure of the micro-reaction chamber. In the process of adjusting the relative size of the pressure values ​​P1 and P2 and observing the sample, since P2=P3, the sample will not be out of focus. It can be understood that at least one through hole may also be provided on the upper partition 3 to connect the reaction chamber and the upper chamber, and then the relative size of the pressure values ​​P1 and P2 is adjusted and the sample is observed. It is also possible to change the thickness of the reaction chamber for observation without losing focus of the sample.

[0069] Figure 5 Show Figure 1 The assembly flow chart of the micro-reaction chamber is shown.

[0070] like Figure 5 As shown, a thicker upper shell 21 (a pressure-resistant film is selected) and a thinner upper separator 3 (a film material is selected) are assembled together, and then the two assembled parts are assembled together in the order of the film toward the center.

[0071] Figure 6 A schematic diagram showing the structure of multiple groups of parallel micro-reaction chambers according to an embodiment of the present disclosure.

[0072] like Figure 6 As shown, the plurality of parallel micro-reaction chambers include a plurality of micro-reaction chambers 1, wherein one or more micro-reaction chambers 1 may be Figure 1 The structure shown.

[0073] The present disclosure also provides a method for adjusting a micro-reaction chamber, comprising the following steps:

[0074] a step of regulating the pressure in the reaction chamber;

[0075] Steps for regulating pressure in the upper chamber and the lower chamber;

[0076] Steps for sample observation.

[0077] In the present disclosure, Figure 1In the micro-reaction chamber shown, in the pressure adjustment step in the reaction chamber, the pressure value P2 in the reaction chamber is adjusted to a predetermined pressure value; in the pressure adjustment step in the upper chamber and / or the lower chamber, the pressure value P3 in the lower chamber is adjusted to the predetermined pressure value, and the pressure value P1 in the upper chamber is adjusted to a value greater than the predetermined pressure value; in the sample observation step, the sample is observed under the conditions of the pressure value P1 in the upper chamber, the pressure value P2 in the reaction chamber, and the pressure value P3 in the lower chamber, which can reduce the influence of the fluid in the reaction chamber on the imaging quality while ensuring that the sample is not out of focus, thereby obtaining a high-resolution image.

[0078] Taking the observation of virus invading cells as an example, the following steps are included:

[0079] The sample cell is placed on the lower separator;

[0080] The pressure in the reaction chamber and the lower chamber is adjusted to atmospheric pressure, and the vertical positions of the upper and lower separators are ensured to be stable while maintaining the natural state of the cells;

[0081] The micro-reaction chamber is placed in the vacuum environment of the TEM, the pressure in the upper chamber is adjusted to be less than the atmospheric pressure, and the distance between the upper and lower separators is increased to allow the virus to fully react with the cells. The pressure in the upper chamber is then adjusted to be greater than the atmospheric pressure, and the distance between the upper and lower separators is reduced to reduce the scattering and absorption of electrons by the liquid.

[0082] TEM is used to observe samples, thereby improving imaging resolution and performing high-quality imaging.

[0083] The above description is only a preferred embodiment of the present disclosure and an explanation of the technical principles used. Those skilled in the art should understand that the scope of the invention involved in the present disclosure is not limited to the technical solution formed by a specific combination of the above technical features, but should also cover other technical solutions formed by any combination of the above technical features or their equivalent features without departing from the inventive concept. For example, the above features are replaced with the technical features with similar functions disclosed in the present disclosure (but not limited to) by each other.

Claims

1. A micro-reaction chamber, characterized in that: include: The shell is provided with an upper separator and a lower separator, which divide the inner cavity of the shell into an upper chamber located at the top, a reaction chamber located in the middle and a lower chamber located at the bottom, and the distance between the upper separator and the lower separator can be adjusted.

2. The micro-reaction chamber according to claim 1, characterized in that: The upper chamber, the reaction chamber, and the lower chamber are all gas chambers; or, the upper chamber and the lower chamber are gas chambers, and the reaction chamber is a liquid chamber.

3. The micro-reaction chamber according to claim 1 or 2, characterized in that: The upper spacer and / or the lower spacer is at least partially a film structure having a thinned area to form an electron beam observation area.

4. The micro-reaction chamber according to claim 3, characterized in that: Part of the upper surface and / or lower surface of the upper separator is thinned in the thickness direction to form the thinned area; and / or, A partial area of ​​the upper surface and / or the lower surface of the lower separator is thinned in a thickness direction to form the thinned area.

5. The micro-reaction chamber according to claim 3, characterized in that: The upper separator and / or the lower separator are composed of an upper layer and a support plate, the upper layer is a membrane structure, the upper layer is thinned along the thickness direction and forms the thinned area with the support plate, or the support plate is thinned along the thickness direction and forms the thinned area with the upper layer, or the upper layer and the support plate are each thinned along the thickness direction to form the thinned area.

6. The micro-reaction chamber according to claim 3, characterized in that: The upper separator and / or the lower separator is / are a non-crystalline carbon film or a polymer film having elasticity.

7. The micro-reaction chamber according to any one of claims 3 to 6, characterized in that: The thickness of the thinned region is 0.3-100 nm, and / or the thickness of the upper separator and / or the lower separator is less than 1 μm, preferably 20-100 nm.

8. The micro-reaction chamber according to any one of claims 4 to 6, characterized in that: The thickness of the upper partition and / or the lower partition is smaller than the thickness of the housing.

9. The micro-reaction chamber according to claim 8, characterized in that: The shell has a thickness of 20-1000 nm, preferably 20-100 nm, and the upper separator and / or the lower separator has a thickness of 0.3-100 nm, preferably 5-20 nm.

10. The micro-reaction chamber according to claim 3, characterized in that: The housing is at least partially a membrane structure.

11. The micro-reaction chamber according to any one of claims 1 to 10, characterized in that: The upper chamber, the reaction chamber and the lower chamber are respectively connected to a pressure control device.

12. The micro-reaction chamber according to claim 11, characterized in that: The pressure value in the reaction chamber is a set value, the pressure value in the upper chamber is greater than, equal to, or less than the set value, and the pressure value in the lower chamber is greater than, equal to, or less than the set value.

13. The micro-reaction chamber according to claim 1, characterized in that: The shell is provided with an observation window to adapt to focusing electron microscopy, optical microscope or X-Ray microscope for sample observation.

14. A plurality of micro-reaction chambers connected in parallel, characterized in that: It comprises a plurality of micro-reaction chambers as described in any one of claims 1-13.

15. A method for adjusting the micro-reaction chamber according to any one of claims 1 to 14, comprising the following steps: a step of regulating the pressure in the reaction chamber; a step of regulating pressure in the upper chamber and / or the lower chamber; Steps for sample observation.

16. The method for adjusting a micro-reaction chamber according to claim 15, characterized in that: In the step of adjusting the pressure in the reaction chamber, the pressure value in the reaction chamber is adjusted to a predetermined pressure value; In the step of adjusting the pressure in the upper chamber and / or the lower chamber, the pressure value in the lower chamber is adjusted to the predetermined pressure value, and the pressure value in the upper chamber is adjusted to a value greater than the predetermined pressure value; In the step of observing the sample, the sample is observed under the conditions of the pressure value in the upper chamber, the pressure value in the reaction chamber, and the pressure value in the lower chamber.

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