A sample displacement stage device and system for in-situ diffraction of biomacromolecule crystals
By using a sample displacement stage device with a linear motor module and a sample well plate clamp in the diffraction of biomacromolecule crystals, in-situ diffraction of samples was achieved, solving the problem of high sample loss rate, improving data acquisition efficiency and accuracy, and reducing experimental costs.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- SHANGHAI ADVANCED RES INST CHINESE ACADEMY OF SCI
- Filing Date
- 2025-08-11
- Publication Date
- 2026-07-03
AI Technical Summary
In the existing technology of biomacromolecule crystal diffraction analysis, the sample loss rate is high, resulting in incomplete or biased data, and increasing the experimental cycle and cost.
A sample displacement stage device consisting of two vertically arranged linear motor modules is used to achieve in-situ diffraction of the sample through the linear motor modules and sample orifice plate fixtures, avoiding damage and positioning deviation of the sample during the operation of the robotic arm.
It significantly improved data acquisition efficiency and quality, reduced sample loss, shortened experimental cycles and reduced costs, while also improving experimental stability and accuracy.
Smart Images
Figure CN224456634U_ABST
Abstract
Description
Technical Field
[0001] This utility model relates to the field of detection instruments, and in particular to a sample displacement stage device and system for in-situ diffraction of biological macromolecular crystals. Background Technology
[0002] Crystallography beamlines are experimental platforms located in synchrotron radiation sources or neutron sources, specifically designed for crystal structure research in fields such as materials (e.g., MOFs, superconductors), chemistry (e.g., catalysis), and biology (e.g., atomic structure determination of proteins, viruses, and nucleic acids). They utilize high-brightness X-rays or neutron beams to perform diffraction analysis on crystal samples to determine atomic-scale structural information. For example, X-ray diffraction, through single-crystal or multi-crystal diffraction techniques, resolves crystal structures (e.g., proteins, inorganic materials, or nanomaterials), which is of great significance for advancing research in life sciences, drug development, and other fields.
[0003] In existing protein diffraction analysis techniques, after crystallization, samples are frozen (e.g., using small molecule immersion techniques), then individually retrieved and rapidly frozen (usually in liquid nitrogen) to form samples in an automated storage system (liquid nitrogen environment). A robotic arm then picks up the samples and loads them onto the diffractometer for subsequent analysis. The robotic arm sample loading process includes sample picking, cryogenic transport, and alignment and positioning. This operation is prone to sample detachment, damage, or positioning errors during robotic arm loading, leading to a high sample loss rate.
[0004] The high sample loss rate, incomplete or biased data due to missing key structures and statistical bias, the need for repeated preparation increases costs and experimental cycles, and waste of synchrotron radiation machine resources and costs.
[0005] How to better reduce sample loss rate has been a long-standing problem in the field of biomolecular diffraction analysis. Utility Model Content
[0006] In view of the shortcomings of the prior art described above, the purpose of this utility model is to provide an operating device and system for in-situ diffraction of biological macromolecular crystals, so as to solve the problem of high sample loss rate in the prior art.
[0007] To achieve the above-mentioned and other related objectives, this utility model is implemented through the following technical solutions.
[0008] This invention provides an operating device for in-situ diffraction of biological macromolecule crystals, wherein the sample shifting stage device includes two mutually perpendicularly arranged linear motor modules:
[0009] An arbitrary linear motor module includes a linear guide rail, a slider, and a control module;
[0010] The slide rail of one linear motor module is fixed to the slider of another linear motor module;
[0011] One linear motor module has a sample orifice plate clamp on its slider; the other linear motor module also has a measuring head connecting and fixing component on the side of its slide rail.
[0012] In one embodiment, the free end of the angle measuring instrument connecting fastener includes an annular connecting portion for fitting onto the angle measuring head.
[0013] In one embodiment, the sample transfer stage device further includes a vertical connection, the vertical connection including a first connection part and a second connection part for auxiliary vertical setting, the first connection part being used to connect to the slider of one of the linear motor modules, and the second connection part being used for the other linear motor module to sit on.
[0014] In one embodiment, the first connecting portion is detachably connected to the slider; and / or, the second connecting portion is detachably connected to the slide rail.
[0015] In one embodiment, the sample well plate clamp has a frame structure for supporting and limiting the sample well plate, wherein one side is used to fix it to the slider.
[0016] In one embodiment, the distance between the sample plate clamp and the slider is adjustable.
[0017] In one embodiment, the sample plate clamp and the slider are fixed by screws, and a plurality of hollow guide posts protrude from its frame structure for the screws to pass through; it also includes locking screws for inserting into the frame from the side to abut against the screws in order to adjust the spacing.
[0018] In one embodiment, the sample plate clamp has two sets of limiting structures arranged in a stepped manner to allow the sample plate to be placed in either direction.
[0019] In one embodiment, the limiting structure is a limiting protrusion or a limiting key, used for interference fit with the side of the sample well plate.
[0020] This utility model also provides a sample displacement stage system, including a sample displacement stage device as described above and a host computer, wherein the host computer is electrically connected to the control module of the linear motor module.
[0021] As described above, the sample displacement stage device and system for in-situ diffraction of biomacromolecule crystals of this invention have the following beneficial effects:
[0022] This invention employs a highly efficient and precise linear motor module and orifice plate clamp to form a sample displacement stage device and system. This sample displacement stage device can be connected and fixed to a goniometer head, allowing for direct in-situ diffraction and crystal analysis of biomolecular samples located in the orifice plate. This eliminates the need to retrieve the sample from the orifice plate and place it in a sample cell for loading with a robotic arm, thus avoiding damage and breakage to the sample during this operation. Using this device and system at a crystallography beamline, in-situ collection of crystals from the crystallography plate at room temperature is achieved. This technology enables the direct acquisition of high-quality diffraction data from the crystallography plate without complex processing. This technical solution can acquire complete diffraction data over a large angular range from a single crystal, significantly improving data acquisition efficiency and quality. Attached Figure Description
[0023] Figure 1 The diagram shown is a three-dimensional schematic of the sample displacement stage device of this utility model.
[0024] Figure 2 Displayed as Figure 1 A magnified structural diagram of point A in the middle.
[0025] Figure 3 The diagram shows the structure of the angle measuring head connection and fixing component.
[0026] Figure 4 The diagram shown is a partial exploded view of the sample displacement stage device, excluding the sample orifice plate fixture.
[0027] Figure 5 The diagram shows the structure of a linear motor module.
[0028] Figure 6 The diagram shows the structure of the sample orifice plate fixture.
[0029] Figure 7 This is one of the side structural schematic diagrams of the sample displacement stage device.
[0030] Figure 8 The diagram shows a front view of a sample displacement stage device in which a sample well plate is fixed in one of the ways.
[0031] Figure 9 Displayed as Figure 8 The AA cross-sectional view is used to show the mating features of the sample orifice plate and the sample orifice plate fixture.
[0032] Figure 10 Displayed as Figure 9 The enlarged structural diagram at point I in the diagram shows the interference fit feature.
[0033] Figure 11The diagram shows a front view of a sample displacement stage device with a sample well plate fixed in an alternative manner.
[0034] Figure 12 Displayed as Figure 11 A BB cross-sectional view is provided to show the mating features of the sample orifice plate and the sample orifice plate fixture.
[0035] Figure 13 Displayed as Figure 12 The enlarged structural diagram at point II in the diagram shows the interference fit feature.
[0036] Figure 14 The diagram shown is a schematic of the system controlling movement in the XY directions to change the hole position in this application.
[0037] Figures 1 to 13 The symbols in the attached icons are explained as follows:
[0038] 1 Linear motor module
[0039] 11, 21 slide rails
[0040] Sliders 12 and 22
[0041] 2 Linear Motor Module
[0042] 3. Sample plate fixture
[0043] 4. Angle measuring head connecting fastener
[0044] 41 Annular connecting part
[0045] 5. Vertical connectors
[0046] 51 First connecting part
[0047] 52 Second connecting part
[0048] 6 Hollow Guide Posts
[0049] 7 Locking screws
[0050] 8 screws
[0051] 9. Limiting Structure
[0052] 10. Hole plate or hole plate simulator Detailed Implementation
[0053] The following specific embodiments illustrate the implementation of this utility model. Those skilled in the art can easily understand other advantages and effects of this utility model from the content disclosed in this specification.
[0054] Please see Figures 1 to 14It should be understood that the structures, proportions, sizes, etc., illustrated in the accompanying drawings are merely for illustrative purposes to aid those skilled in the art and are not intended to limit the scope of this invention. Therefore, they have no substantial technical significance. Any modifications to the structure, changes in proportions, or adjustments to size, without affecting the effectiveness and purpose of this invention, should still fall within the scope of the disclosed technical content. Furthermore, the terms "upper," "lower," "left," "right," "middle," and "one" used in this specification are merely for clarity and not intended to limit the scope of this invention. Changes or adjustments to their relative relationships, without substantially altering the technical content, should also be considered within the scope of this invention.
[0055] In crystallography beamlines, especially single-crystal diffraction beamlines using synchrotron or neutron sources, the sample displacement stage is a crucial component. Its primary task is to precisely position, orient, and move the sample (such as a crystal) to the center of the X-ray / neutron beam spot, and to adjust the crystal's orientation as needed during the experiment. Precise positioning is a prerequisite for obtaining high-quality diffraction data. During data acquisition, the sample displacement stage must possess high mechanical stability to ensure that the sample position does not shift even slightly due to environmental vibrations or thermal drift. Any minute vibration can lead to blurred diffraction points and decreased resolution.
[0056] like Figure 1 As shown, this utility model provides a sample displacement stage device for in-situ diffraction of biomacromolecule crystals, comprising two mutually perpendicular linear motor modules.
[0057] An arbitrary linear motor module includes a slide rail, a slider, and a control module;
[0058] The slide rail of one linear motor module is fixed to the slider of another linear motor module;
[0059] One linear motor module has a sample orifice plate clamp on its slider; the other linear motor module also has a measuring head connecting and fixing component on the side of its slide rail.
[0060] In this application, the angle measuring head connecting fastener is used to fix the entire sample displacement stage device to the angle measuring head, thereby facilitating subsequent operations on the orifice plate held and fixed in the sample orifice plate fixture of the sample displacement stage device for in-situ X-ray testing. The angle measuring head connecting fastener can be detachably connected to the angle measuring head of the diffractometer by means of sleeve, threaded connection, or snap-fit. In a... Figure 3In a more specific embodiment shown, the angle measuring head connecting fastener includes an annular connecting portion for sleeved onto the angle measuring head. The annular connecting portion is detachably connected to the angle measuring head, such as by snap-fit, sleeve, or screw connection. Generally, this annular connecting portion is used to detachably sleeve onto the corresponding position of the angle measuring head of the X-ray diffractometer. The corresponding position has an axial structure, thereby fixing the entire sample displacement stage device onto the angle measuring head by snap-fit, adhesive, or screw fixation after sleeve. As the angle measuring head rotates, it drives the rotation of the sample displacement stage device and the sample in the clamped test plate.
[0061] Biological sample well plates are generally used to store samples and are available in 96-well and 384-well configurations. These plates can have round or square holes. The sample displacement stage device claimed in this application can directly hold the well plate using a clamp and switch between different holes using a linear motor module. The rotation of the goniometer head drives the sample displacement stage device to rotate, thereby achieving efficient and low-sample-damage in-situ X-ray crystal diffraction analysis. In one specific embodiment, the goniometer head can rotate 360°, which can also drive the sample well plate to rotate, thereby changing the angle and obtaining more information about the sample.
[0062] The linear motor module used in this application is part of the prior art. It is based on direct electromagnetic thrust drive, eliminating the need for intermediate transmission mechanisms; it can achieve efficient and high-precision linear motion. The control module includes a motion control module, which receives controller commands, adjusts the primary current (controls the primary speed), and combines position information from detection feedback elements to adjust the current in real time for more precise control; generally, it also contains a primary (mover), a secondary (stator), and detection feedback elements (such as Hall sensors). The linear guide rail and slider serve as the supporting and guiding mechanism.
[0063] The control module is used to receive instructions from the host computer and convert them into motor displacement parameters (such as current signals) to control the motor movement.
[0064] In one embodiment, such as Figure 1 and Figure 4As shown, the sample shifting stage device described in this application further includes a vertical connector 5 for assisting in vertical setting. The vertical connector 5 includes a first connecting part 51 and a second connecting part 52, which are vertically set. The first connecting part 51 is used to connect to the slider 12 of one of the linear motor modules 1, and the second connecting part 52 is used for mounting another linear motor module 2. Specifically, the slider 12 and the first connecting part 51 are detachably connected, such as by snap-fit or threaded connection. In a specific embodiment shown in the figure, the slider 12 and the first connecting part 51 are detachably connected by screw holes and screws. Specifically, another linear motor module 2 is mounted on the second connecting part 52, and the slide rail of the linear motor module is detachably connected to the second connecting part 52, such as by snap-fit or threaded connection. In a specific embodiment shown in the figure, the slide rail 21 of the linear motor module 2 is detachably connected to the second connecting part 52 by threaded holes and screws. With the help of the vertical connector 5, the movement of the first motor module is perpendicular to the movement direction of the second motor module, or its slide rail is perpendicular, and the two sliders can move independently, with their independent movement directions also perpendicular, such as along the X and Y directions respectively. Therefore, the precise control of the perforated plate displacement in the X and Y directions by the linear motor module allows for the switching of different perforation samples.
[0065] In a like Figure 1 and Figure 6 In the illustrated embodiment, the sample well plate clamp 3 has a frame structure for supporting and positioning the sample well plate. In one embodiment, one side of the sample well plate clamp is used to fix it to the slider, as shown in the figure, and can be detachably connected by screws. Figure 1 and Figure 6 As shown, it specifically has a rectangular frame structure, or it could be a frame structure adapted to a perforated plate. Further, as... Figure 6 As shown, the frame structure forms a placement stage for inserting or removing the sample well plate from the side. Figure 6 The direction indicated by the middle arrow is the sample plate insertion direction. This placement stage can be a stepped stage formed by a frame structure, and the side in the insertion direction only has a platform, without any stepped elevation.
[0066] like Figure 1 and Figure 2 In one embodiment shown, the distance between the sample plate clamp 3 and the slider 22 is adjustable. For example, the distance can be 1–8 mm. Figure 2 In the embodiment shown, the sample plate clamp 3 and the slider 22 are fixed by screws 8, and a plurality of hollow guide posts 6 protrude from its frame structure for screws to pass through; it also includes locking screws 7, which are inserted into the frame from the side to abut against the screws 8 in order to adjust the spacing. Figure 2 The direction indicated by the middle arrow is the adjustable position direction (such as forward or backward adjustment) along this direction or its opposite direction, thereby adjusting the spacing.
[0067] The sample well plate clamp with a frame structure in this application also forms a limiting structure to engage, limit, and fix the inserted sample well plate. The limiting structure protrudes and is positioned relative to each other. To facilitate installation, disassembly, and operation, the sample well plate clamp provided in this application can hold the sample well plate in two orientations. In a... Figure 6 In the specific embodiment shown, the sample well plate clamp 3 has two sets of limiting structures 9 arranged in a stepped layout, allowing the sample well plate to be placed in either direction. Specifically, the two sets of limiting structures are respectively located at corresponding positions on the two stepped placement platforms. Each set of limiting structures 9 includes two oppositely arranged limiting protrusions, a limiting key, a snap-fit key, or an interference fit key. Figure 6 It can be seen that the two sets of limiting structures are arranged in an upper and lower layer (set on the lower step placement platform and the upper step placement platform respectively); more specifically, as Figure 6 As shown, the lower limiting structure includes four limiting structures arranged in pairs opposite each other; the upper limiting structure also includes four limiting structures arranged in pairs opposite each other. The upper and lower limiting structures are staggered. More specifically, in the upper and lower layers, one pair of limiting structures is positioned closer to the insertion end, and the other pair is positioned further away from the insertion end. Specifically, the limiting structure is a limiting protrusion or a limiting key, used for interference fit with the side of the sample well plate; see [link to details]. Figure 10 and Figure 13 .
[0068] To better demonstrate that the sample well plate fixture in this application can place the sample well plate in two directions, combined with Figures 7-13 Further explanation is provided. The sample well plate with two opposing orientations in this application allows for placement of the well plate either forward or backward, thus enabling the goniometer to obtain more diffraction resolution information from the sample without requiring a larger rotation angle. For example, Figure 7 The diagram shown is a structural schematic of a mold without or without a perforated plate, for comparison. Figures 8-10 (placed square) and Figures 11-13 (Upside down)
[0069] For example Figure 8 As shown, the orifice plate 10 (e.g., a 96-well plate) is inserted into the sample orifice plate fixture from the side and is supported and confined on the fixture. By controlling the movement of two linear motor modules via a host computer, the orifice plate can be moved together, thereby changing the sample orifice for in-situ analysis. For example, controlling the linear motor modules to move in the XY direction to a specific orifice position allows for in-situ diffraction analysis of the sample at that orifice position. Specifically, the orifice plate 10 is engaged and fixed by the lower limiting structure of the sample orifice plate fixture 3, with the interference fit forming as shown in the figure. Figure 11 As shown.
[0070] For example Figure 11 and 12 As shown, the well plate 10 (e.g., a 96-well plate) is inserted from the side of the sample well plate fixture in the reverse direction and is supported and confined on the sample well plate fixture. By controlling the movement of two linear motor modules via a host computer, the well plate can be moved together, thereby changing the sample well position for in-situ analysis. For example, controlling the linear motor module to move to a specific well position in the XY direction allows for in-situ diffraction analysis of the sample at that well position. Specifically, the well plate 10 is locked and fixed by the upper limiting structure of the sample well plate fixture 3, and the resulting interference fit is shown in the figure. Figure 13 As shown.
[0071] This utility model also provides a sample displacement stage system, including the above-mentioned sample displacement stage device and a host computer, wherein the host computer is connected to the motor drive electrical signal.
[0072] In one embodiment, the host computer in this application can be an industrial computer, a personal computer, a server, an embedded industrial control device, or a cloud computing platform.
[0073] The specific process of control in this application is described as follows:
[0074] The host computer sends control commands;
[0075] The control module of the sample displacement stage receives the instruction and converts it into motor displacement parameters (current signal). Simultaneously, it sends drive commands to the first and second motors to generate the specified displacement; this causes the slider, fixture, and orifice plate to move, with the sample orifice plate moving along the XY direction. The XY directions are perpendicular to each other.
[0076] After the sample plate has moved, the control module sends the displacement information to the host computer.
[0077] The host computer records and refreshes the current position of the sample aperture plate based on the received displacement information, thereby obtaining the sample information of the currently aligned beam.
[0078] In the above technical solution of this application, the length and height of the orifice plate are both fixed.
[0079] In one specific embodiment, considering the limited space at the sample cell of the diffractometer, the linear motor module selected in this application is characterized by high efficiency, precision, and portability. To allow the aforementioned device to be placed in the available space, the track length of the linear motor module in the X or Y direction can be controlled. In one specific embodiment, the track length of the linear motor module can be approximately 10 cm. For example, for a 96-well plate, the process can be found in [reference needed]. Figure 13As shown, the current hole position can be confirmed as blue E5 by the host computer. If you want to change the hole position, you can set the X-axis movement step size (distance) and Y-axis movement step size (distance) through the host computer. After receiving the instruction, the control module will drive the control slider to move precisely along the X or Y direction by the set step size, thereby achieving precise hole position replacement. Since the length and height of the perforated plate and the size of the holes, as well as the distance between two adjacent holes, are all fixed, this step size can be set by inputting it at the beginning, such as setting the center distance between two adjacent holes as one step size.
[0080] In one embodiment, the control module can maintain the position even after power failure using an absolute value method. This is achieved through an absolute encoder or similar technology, ensuring that the control module accurately retains the current position of the sample plate upon power restoration after a power outage, without requiring recalibration or zeroing.
[0081] In summary, the automated sample displacement stage device and system enable the individual testing of all samples on a well plate (such as a 96-well plate) with only a single installation, significantly improving experimental efficiency. Furthermore, because all samples are loaded in a single setup, the device and control system ensure the accurate positioning of each sample through precise control of the motor movement. In addition, the device guarantees sufficiently high stability when connected to crystallography beamlines, ensuring that the samples remain continuously within the X-ray path during the experiment. While maintaining stability, the device also remains within the load-bearing capacity of a high-precision diffractometer.
[0082] The technical solution employed in this invention successfully solves some key technical challenges in traditional crystal diffraction methods, particularly in in-situ data acquisition and applications within a wide diffraction range. By achieving in-situ collection of crystals from a crystallography plate at room temperature using a crystallography beamline, this technique enables the direct acquisition of high-quality diffraction data from the crystallography plate without complex processing. This technical solution can acquire complete diffraction data over a wide angle range from a single crystal, significantly improving data acquisition efficiency and quality. Specifically, based on this technical solution, diffraction data for proteins such as lysozyme and Pin1 at room temperature can be successfully acquired, and their three-dimensional structures can be resolved smoothly. In this way, a single crystal can complete the complete diffraction data acquisition without the need for repeated crystal preparation processes, thereby greatly shortening the experimental cycle and reducing experimental costs.
[0083] Furthermore, the system boasts high compatibility, being compatible with multiple crystallography beamlines (such as BL17B1, BL18U1, and BL19U1), further expanding its application scope. This technology not only improves the efficiency and accuracy of data acquisition but also provides a more efficient and convenient technical means for fields such as protein structure analysis, possessing significant technical and economic value.
[0084] The above embodiments are merely illustrative of the principles and effects of this utility model and are not intended to limit the scope of this utility model. Any person skilled in the art can modify or alter the above embodiments without departing from the spirit and scope of this utility model. Therefore, all equivalent modifications or alterations made by those skilled in the art without departing from the spirit and technical concept disclosed in this utility model should still be covered by the claims of this utility model.
Claims
1. A sample displacement stage apparatus for in situ diffraction of biological macromolecular crystals, characterized by, The sample shifting stage device includes two linear motor modules arranged perpendicularly to each other: An arbitrary linear motor module includes a linear guide rail, a slider, and a control module; The slide rail of one linear motor module is fixed to the slider of another linear motor module; One linear motor module has a sample orifice plate clamp on its slider; the other linear motor module also has a measuring head connecting and fixing component on the side of its slide rail.
2. The sample displacement stage apparatus of claim 1, wherein, The free end of the angle measuring head connecting fastener includes an annular connecting part for fitting onto the angle measuring head.
3. The sample displacement stage apparatus of claim 1, wherein, The sample transfer stage device further includes a vertical connector for assisting in vertical setting. The vertical connector includes a first connecting part and a second connecting part that are vertically set. The first connecting part is used to connect to the slider of one of the linear motor modules, and the second connecting part is used to allow another linear motor module to sit on it.
4. The sample displacement stage apparatus of claim 3, wherein, The first connecting part is detachably connected to the slider; and / or, the second connecting part is detachably connected to the slide rail.
5. The sample displacement stage apparatus of claim 2, wherein, The sample well plate clamp has a frame structure and is used to support and limit the position of the sample well plate.
6. The sample displacement stage apparatus of claim 5, wherein, The distance between the sample plate clamp and the slider is adjustable; and / or, the frame structure is formed with a placement stage for inserting or removing the sample plate from the side.
7. The sample displacement stage apparatus of claim 6, wherein, The sample well plate fixture is fixed to the slider by screws, and its frame structure has several hollow guide posts protruding to allow the screws to pass through; it also includes locking screws, which are inserted into the frame from the side to abut against the screws in order to adjust the spacing.
8. The sample displacement stage apparatus of claim 1, wherein, The sample well plate clamp has two sets of limiting structures arranged in a stepped manner to allow the sample well plate to be placed in either direction.
9. The sample displacement stage apparatus of claim 8, wherein, The limiting structure is a limiting protrusion or a limiting key, used for interference fit with the side of the sample well plate.
10. A sample displacement stage system, characterized in that, It includes the sample displacement stage device and the host computer as described in any one of claims 1 to 9, wherein the host computer is electrically connected to the control module of the linear motor module.