Isotope calcium 45 uptake quantitative analysis experimental device and experimental method thereof

Abstract

This invention relates to an experimental apparatus and method for quantitative analysis of calcium-45 isotope uptake, belonging to the field of cell biology experimental equipment. The apparatus includes a negative pressure chamber, an experimental cover, assembled filter plates, control valves, and an experimental filter membrane. The negative pressure chamber has a storage cavity inside, with the experimental cover sealed at the top opening. The experimental cover has multiple arrayed flow channels, each connected to the storage cavity. An assembled filter plate is sealed within each flow channel, with its top surface lower than the top surface of the experimental cover to form a groove, preventing reaction liquid overflow. An experimental filter membrane is laid on the upper surface of the assembled filter plate. Each flow channel has an independent control valve for individual on / off control. The apparatus also includes a radiation baffle to block beta radiation. This apparatus enables multi-channel independent filtration, effectively preventing cross-contamination and improving experimental efficiency. It also possesses advantages such as radiation protection, compact structure, and safe operation.

Description

Technical Field

[0001] This invention relates to the field of experimental equipment for cell biology. In particular, it relates to an experimental apparatus and method for quantitative analysis of calcium-45 isotope uptake. Background Technology

[0002] Calcium signaling is widely involved in various life activities. Calcium ion concentrations vary greatly among different organelles; the cytoplasmic calcium ion concentration is approximately 100 nM, while the endoplasmic reticulum (ER) calcium ion concentration is approximately 0.5-1 mM, a difference of tens of thousands of times. ER calcium homeostasis is maintained by the SERCA pathway, which continuously pumps calcium ions back to the ER, and ER calcium release channels. Alterations in SERCA activity are closely related to various diseases, and accurate analysis of SERCA activity is crucial for identifying drugs or proteins that regulate SERCA activity.

[0003] Currently, there is no specialized equipment capable of quantitatively measuring the activity of the endoplasmic reticulum calcium pump SERCA via calcium-45 uptake. Existing experimental methods typically employ commercially available ordinary vacuum filter manifolds, which have the following drawbacks: first, they lack protection against beta rays, posing a risk of radiation exposure to operators; second, they lack dedicated storage devices for radioactive sources and scintillation tubes, making radioactive material management inconvenient; third, in multi-channel filtration, each channel cannot be independently controlled, resulting in poor operational flexibility and low experimental efficiency; and fourth, the entire experimental process relies on dispersed general-purpose equipment, leading to low integration and cumbersome operation. Therefore, there is an urgent need for a specialized experimental apparatus and supporting method for the quantitative analysis of calcium-45 uptake. Summary of the Invention

[0004] The purpose of this invention is to provide an experimental apparatus and method for quantitative analysis of calcium-45 isotope uptake, in order to solve one or all of the technical problems existing in the prior art.

[0005] To achieve the above objectives, the present invention provides the following solution: This invention provides an experimental apparatus for quantitative analysis of calcium-45 isotope uptake, comprising a negative pressure chamber, an experimental cover, an assembled filter plate, a control valve, and an experimental filter membrane. The negative pressure chamber has a storage cavity on its inner side and an opening at its top. The side walls of the negative pressure chamber are provided with a negative pressure connector and a drain connector communicating with the storage cavity. The experimental cover is fixedly installed at the opening at the top of the negative pressure chamber, and the connection between the experimental cover and the negative pressure chamber is sealed. A through-flow groove is provided on the experimental cover, communicating with the storage cavity. The control valve is located on the experimental cover and cooperates with the flow groove, controlling the opening and closing of the flow groove. The assembled filter plate is placed inside the flow groove, and the connection between the assembled filter plate and the flow groove is sealed. The experimental filter membrane is laid flat on the upper surface of the assembled filter plate.

[0006] According to one embodiment of the present invention, the mounting height of the filter plate is lower than the horizontal height of the upper end face of the experimental cover, so that the mounting filter plate and the flow channel form a groove for storing experimental materials.

[0007] According to one embodiment of the present invention, multiple flow channels are provided, and an array of multiple flow channels is arranged on the experimental cover; each flow channel is provided with an assembly filter plate, a control valve and an experimental filter membrane, and the opening and closing of a single flow channel can be controlled individually by the corresponding control valve.

[0008] According to one embodiment of the present invention, the experimental cover is provided with a horizontal limiting groove, the limiting groove passing through the flow groove; the control valve includes a cylindrical valve body and an adjusting head, the valve body is provided with a valve port, the valve body is rotatably embedded in the limiting groove, and the valve port is located at the flow groove; rotating the valve body can adjust the communication state between the valve port and the flow groove; the adjusting head is located on the outside of the experimental cover, and rotating the adjusting head can drive the valve body to rotate.

[0009] According to one embodiment of the present invention, the adjusting head is provided with a marking line, which is used to indicate the opening direction of the valve port.

[0010] According to one embodiment of the present invention, the assembled filter plate is a quartz sand filter plate.

[0011] According to one embodiment of the present invention, it further includes a radiation baffle, which is mounted on the side of the negative pressure chamber; the upper part of the radiation baffle is bent to form an observation section.

[0012] According to one embodiment of the present invention, the negative pressure connector is disposed on the upper part of the side wall of the negative pressure box to prevent the liquid in the storage cavity from being discharged through the negative pressure connector; the drain connector is disposed on the bottom of the side wall of the negative pressure box for draining the liquid in the storage cavity.

[0013] A quantitative analysis method for calcium-45 uptake using isotope, employing the aforementioned quantitative analysis apparatus for calcium-45 uptake, includes the following steps: S1. After wetting the experimental filter membrane, lay it flat on the upper surface of the assembled filter plate placed in the flow channel, and close the control valve; S2. Incubate the reaction solution containing endoplasmic reticulum vesicles and calcium 45 isotope at the set calcium ion concentration and time. S3. After the incubation is completed, the reaction solution is transferred to the filter membrane at the top of the filter plate in the flow channel. The control valve is opened and the negative pressure is started so that the reaction solution enters the storage chamber after being filtered through the experimental filter membrane and the filter plate. The endoplasmic reticulum vesicles are trapped on the experimental filter membrane. S4. Close the control valve and repeat steps S1 to S3 to process other samples in batches. S5. After the experimental filter membrane dries, transfer it to a scintillation tube, add scintillation fluid, and perform isotope concentration detection.

[0014] According to one embodiment of the present invention, in step S3, the reaction solution is transferred in its entirety to the flow channel for filtration, and in step S5, the experimental filter membrane is dried and then transferred as a whole to the scintillation tube.

[0015] Beneficial effects This invention has at least one of the following technical effects: First, this invention creates a groove for storing experimental materials by setting multiple arrayed flow channels on the experimental cover and setting the height of the assembled filter plate in each flow channel to be lower than the upper end face of the experimental cover. This allows operators to directly transfer the reaction liquid into the groove for temporary storage during multi-channel batch filtration, avoiding liquid overflow or cross-contamination due to surface tension. At the same time, the reaction liquid is effectively constrained in the groove area during negative pressure filtration, improving the stability of multi-channel parallel operation and the reliability of experimental results.

[0016] Secondly, by configuring an independently controlled valve for each flow cell, the channels do not interfere with each other. Operators can flexibly choose to start or close any channel according to the experimental design. When processing samples with different calcium ion concentrations or at different time points in batches, there is no need to repeatedly disassemble and reassemble the filter membrane or switch pipelines, which significantly improves experimental efficiency. It is especially suitable for SERCA activity determination experiments that require the filtration of multiple samples in a short time. Its operational flexibility and high throughput advantages are outstanding.

[0017] Third, the present invention installs a radiation baffle on the side of the negative pressure chamber. The baffle is made of 1 cm thick transparent acrylic material and has a bending observation part. It can effectively block the β rays released by calcium 45 to protect the operator from radiation damage, without obstructing the operator's line of sight, so as to facilitate real-time observation of the filtration process and the status of the filter membrane. At the same time, the radiation baffle has a simple structure, is easy to clean, and is integrated with the negative pressure chamber, which improves the safety and convenience of the experiment.

[0018] Fourth, by placing the negative pressure connector on the upper part of the side wall of the negative pressure box and the drain connector on the bottom of the side wall, the present invention makes reasonable use of the gravity of the liquid and the direction of the negative pressure airflow, effectively preventing the waste liquid in the storage chamber from being sucked back into the negative pressure pump through the negative pressure connector, thus avoiding equipment pollution and damage. The drain connector is located at the bottom to facilitate the complete discharge of waste liquid. Combined with the sealing structure of the negative pressure box, the entire negative pressure system operates stably, is easy to maintain, and extends the service life of the device. Attached Figure Description

[0019] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0020] Figure 1 A schematic diagram of the overall structure of the experimental apparatus for quantitative analysis of calcium-45 isotope uptake. Figure 2 for Figure 1 Enlarged structural diagram at point A; Figure 3 A side cross-sectional view of the experimental setup for quantitative analysis of calcium-45 isotope uptake. Figure 4 Schematic diagram of the control valve status in the experimental apparatus for quantitative analysis of calcium-45 isotope uptake. Figure 1 ; Figure 5 Schematic diagram of the control valve status in the experimental apparatus for quantitative analysis of calcium-45 isotope uptake. Figure 2 ; Figure 6 A cross-sectional view of the control valve section in the experimental apparatus for quantitative analysis of calcium-45 uptake from isotope. Figure 7 A schematic diagram showing the setup of the radiation baffle in the experimental apparatus for quantitative analysis of calcium-45 uptake. Figure 8 A schematic diagram showing the setup of the negative pressure pump and waste bottle in the experimental apparatus for quantitative analysis of calcium-45 uptake. Figure 9 A structural block diagram of the experimental method for quantitative analysis of calcium-45 isotope uptake.

[0021] Explanation of reference numerals in the attached figures: Negative pressure box 1, storage chamber 11, negative pressure connector 12, drain connector 13, experimental cover 2, limiting groove 21, assembled filter plate 3, control valve 4, valve body 41, valve port 411, adjusting head 42, marking line 421, experimental filter membrane 5, radiation baffle 6. Detailed Implementation

[0022] The features and exemplary embodiments of various aspects of the present invention will now be described in detail. To make the objectives, technical solutions, and advantages of the present invention clearer, the present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments. It should be understood that the specific embodiments described herein are only configured to explain the present invention and to exemplify the principles of the present invention, and are not configured to limit the present invention. In addition, the structural components in the drawings are not necessarily drawn to scale. For example, the dimensions of some structural components or regions in the drawings may be enlarged for other structural components or regions to aid in the understanding of the embodiments of the present invention.

[0023] The directional terms used in the following description refer to the directions shown in the figures and are not intended to limit the specific structure of the embodiments of the present invention. In the description of the present invention, it should be noted that, unless otherwise stated, the terms "installation," "connection," and "joining" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral connection; they can refer to a direct connection or an indirect connection through an intermediate medium. Those skilled in the art can understand the specific meaning of the above terms in the present invention according to the specific circumstances.

[0024] Furthermore, the terms "comprising," "including," "having," or any other variations thereof are intended to cover non-exclusive inclusion, such that a structure or component that includes a list of elements includes not only those elements but also other structural elements that are not expressly listed or inherent to the structure or component. Without further limitations, an element defined by the phrase "comprising..." does not exclude the presence of other identical elements in the article or apparatus that includes the element.

[0025] Spatial relation terms such as "below," "under," "under," "low," "above," "on," and "high" are used for descriptive convenience to explain the positioning of one element relative to a second element, indicating that these terms are intended to cover different orientations of the device, in addition to those different from those shown in the figure. Furthermore, phrases such as "one element on / below another element" can indicate that two elements are in direct contact, or that there are other elements between the two elements. In addition, terms such as "first" and "second" are also used to describe individual elements, areas, parts, etc., without specifically indicating order or sequence, and should not be considered restrictive. Similar terms are used throughout the description to represent similar elements.

[0026] It will be apparent to those skilled in the art that the present invention can be practiced without requiring some of these specific details. The following description of embodiments is merely intended to provide a better understanding of the invention by illustrating examples of the invention.

[0027] In the following embodiments, there may be descriptions such as "this device". Those skilled in the art should understand that "this device" refers to the experimental apparatus and experimental method for quantitative analysis of calcium-45 isotope uptake provided by the present invention.

[0028] An experimental apparatus for quantitative analysis of calcium-45 isotope uptake.

[0029] The negative pressure chamber 1 is made of transparent acrylic sheet with a thickness of 1 cm, which can block beta rays and facilitate observation of the internal liquid level. The inner side of the negative pressure chamber 1 forms a storage cavity 11 for temporarily storing the filtered waste liquid. The top of the negative pressure chamber 1 is open, and an experimental cover 2 is installed at the open. The experimental cover 2 and the negative pressure chamber 1 are sealed together by a sealing ring to ensure that the negative pressure does not leak.

[0030] The negative pressure connector 12 is located on the upper side wall of the negative pressure chamber 1, near the top, and is used to connect to an external negative pressure pump. Because the negative pressure connector 12 is located high, even if a certain amount of waste liquid accumulates in the storage chamber 11, the waste liquid will not be sucked into the negative pressure pump through the negative pressure connector 12, thus protecting the negative pressure pump from contamination. The drain connector 13 is located at the bottom side wall of the negative pressure chamber 1 and is used to connect to a waste liquid bottle or a drain pipe. After the experiment is completed or when too much liquid accumulates in the storage chamber 11, the drain connector 13 can be opened to drain the waste liquid.

[0031] The experimental cover 2 is provided with multiple flow channels. In this embodiment, there are 14 flow channels, which are evenly distributed on the experimental cover 2 in an array of 2 rows and 7 columns. Each flow channel is a circular through hole that penetrates the experimental cover 2. Its lower end is connected to the storage cavity 11 of the negative pressure box 1, and its upper end is used to receive the reaction liquid.

[0032] Each flow channel is equipped with an assembly filter plate 3, which is a quartz sand filter plate. The preparation method is as follows: quartz sand particles with a particle size of 0.3-0.8 mm are mixed with a small amount of binder, loaded into a mold, and sintered at a high temperature of 1200-1300 degrees Celsius to form a porous plate with a large number of interconnected micropores. The thickness of the quartz sand filter plate is about 5 mm, and its porosity is about 30-40%. It can support the experimental filter membrane 5 above it and allow the liquid to pass through quickly under negative pressure. The outer edge of the assembly filter plate 3 is sealed with the inner wall of the flow channel by a silicone sealing ring to prevent liquid from leaking from the edge.

[0033] The top surface of the assembled filter plate 3 is set at a height approximately 3 mm lower than the upper end surface of the experimental cover 2, thereby forming a shallow groove between the assembled filter plate 3 and the side wall of the flow channel. This groove is used to temporarily store the reaction liquid before filtration, preventing the reaction liquid from overflowing the flow channel due to surface tension.

[0034] Each flow channel is equipped with a corresponding control valve 4. The specific structure of the control valve 4 is as follows: A horizontal limiting groove 21 is opened on the experimental cover 2 in a direction perpendicular to the flow channel. The limiting groove 21 is a cylindrical hole with a diameter slightly larger than the diameter of the valve body 41 of the control valve 4. The valve body 41 of the control valve 4 is cylindrical and made of polytetrafluoroethylene (PTFE), which has excellent corrosion resistance and self-lubricating properties. A through valve port 411 is opened on the valve body 41, and the diameter of the valve port 411 is the same as the diameter of the flow channel. The valve body 41 is rotatably installed in the limiting groove 21, and the position of the valve port 411 corresponds to the flow channel. One end of the valve body 41 extends out of the experimental cover 2 and is fixedly connected to an adjusting head 42. 2 is a knob shape, easy to grip and rotate with fingers. The surface of the adjusting head 42 is engraved with a marking line 421. The direction of the marking line 421 is consistent with the axis of the valve port 411. When the operator rotates the adjusting head 42 so that the marking line 421 points to the axis of the flow channel, the valve port 411 is completely aligned with the flow channel. At this time, the flow channel is in a passable state, and negative pressure can be applied to the reaction liquid above the assembled filter plate 3. When the adjusting head 42 is rotated so that the marking line 421 is perpendicular to the axis of the flow channel, the valve port 411 and the flow channel form a cross, and the solid part of the valve body 41 blocks the flow channel. At this time, the flow channel is in a blocked state. In this way, each channel can be independently controlled to open and close without interfering with each other.

[0035] Each flow channel has an experimental filter membrane 5 laid on the upper surface of the assembly filter plate 3. The experimental filter membrane 5 is a nitrocellulose filter membrane or a mixed cellulose filter membrane with a pore size of 0.45 micrometers. Its diameter is slightly larger than that of the assembly filter plate 3. After being laid flat, it can completely cover the upper surface of the assembly filter plate 3, and its edges are pressed by the side wall of the flow channel.

[0036] A radiation baffle 6 is mounted on the front of the negative pressure chamber 1. The baffle 6 is also made of 1 cm thick transparent acrylic sheet. The lower part of the baffle 6 is vertically fixed to the workbench or connected to the base of the negative pressure chamber 1, while the upper part is bent backward to form an inclined observation section. The angle between the observation section and the vertical direction is approximately 30 degrees. When the operator is using the equipment, the radiation baffle 6 is positioned precisely between their body and the negative pressure chamber 1, blocking any beta rays that may be scattered from the direction of the negative pressure chamber 1. Simultaneously, the operator can directly observe the status of each channel on the experimental cover 2 at the top of the negative pressure chamber 1 through the observation section, facilitating operation.

[0037] The isotope calcium-45 uptake quantitative analysis experimental apparatus described above in this embodiment is used to quantitatively analyze the SERCA calcium pump activity in test samples containing endoplasmic reticulum vesicles. The specific operating steps are as follows. S1: Installation of experimental filter membrane 5. Use a suitable size experimental filter membrane 5 and wet it thoroughly with washing buffer. Open the control valve 4 of a certain channel on the negative pressure box 1. Lay the wetted experimental filter membrane 5 flat on the upper surface of the filter plate 3 in that channel, ensuring that the filter membrane completely covers the filter plate 3 and that the edges are flat and free of air bubbles. Then rotate the adjusting head 42 of that channel so that the marking line 421 is perpendicular to the axis of the flow channel. Close the control valve 4. In the same way, install the wetted experimental filter membrane 5 in multiple channels according to the number of samples required for the experiment, and keep the control valve 4 of all channels closed.

[0038] S2: Incubation of the reaction solution. Prepare a reaction solution containing endoplasmic reticulum vesicles and calcium-45 isotope. The reaction solution contains: a suspension of the test sample, calcium-45 isotope, oxalate, ATP, a buffer system, and a set concentration of free calcium ions. The calcium ion concentration can be set to multiple gradients according to experimental needs, such as from 0.1 μM to 10 μM, to cover the physiological response range of SERCA. Place the prepared reaction solution in a reaction tube and put it in a constant temperature water bath. Keep it warm for the set incubation time, which can be set according to experimental needs, such as 30 seconds, 60 seconds, or 90 seconds. For each calcium ion concentration and each time point, an independent reaction tube is usually set up. At the same time, a blank control tube is set up to subtract background non-specific binding.

[0039] S3: After filtration and incubation, immediately remove the reaction tube from the water bath and quickly transfer the entire reaction solution to the flow channel of the corresponding channel. Since the height of the assembled filter plate 3 is lower than the upper surface of the experimental cover 2, a groove is formed between the assembled filter plate 3 and the flow channel, and the reaction solution is temporarily contained in the groove. Then, rotate the adjusting head 42 of the channel so that the marking line 421 on the adjusting head 42 points to the axis of the flow channel, open the control valve 4, and start the negative pressure pump at the same time so that the negative pressure is transmitted to the storage chamber 11 and the bottom of the flow channel through the negative pressure connector 12. Under the action of negative pressure, the reaction solution quickly passes through the experimental filter membrane 5. The liquid part flows into the storage chamber 11 through the assembled filter plate 3, while the endoplasmic reticulum vesicles in the reaction solution are trapped on the upper surface of the experimental filter membrane 5. After the reaction solution is completely drained, rotate the adjusting head 42 to close the control valve 4. For samples that require different time points or different calcium concentration conditions, repeat the above filtration operation using different channels.

[0040] S4: Batch processing of other samples. After closing the control valve 4 of the current channel, replace the experimental filter membrane 5 of the channel. Then, operate the next sample according to steps S1 to S3. Since the device of the present invention has multiple arrayed flow channels and independent control valves 4, the operator can process multiple samples under different conditions sequentially or in parallel according to the experimental design. Each channel operates and is controlled independently without interference. Through batch processing, the filtration and separation of dozens of samples can be completed in a short time, which significantly improves experimental efficiency.

[0041] S5: Drying, Transfer and Detection. After all samples have been filtered, remove each experimental filter membrane 5 from the top of the assembled filter plate 3. Place the filter membrane on clean absorbent paper or filter paper and allow it to dry naturally at room temperature for about 10-15 minutes until there is no obvious moisture on the surface of the filter membrane. Take scintillation tubes corresponding to the number of experimental filter membranes 5. Add an appropriate amount of scintillation fluid to each scintillation tube in advance. Transfer the dried experimental filter membrane 5 into the scintillation tube, so that the filter membrane is completely immersed in the scintillation fluid. Tighten the cap and gently shake to ensure that the filter membrane and scintillation fluid are in full contact. Place the scintillation tube into the liquid scintillation counter, set an appropriate counting time, and perform isotope concentration detection, recording the count per minute.

[0042] Data Analysis: By subtracting the background from the count value of the blank control, the net count of each sample was obtained. The net count was converted into calcium ion uptake using standard samples. For calcium uptake at different time points under each calcium ion concentration, a linear regression was performed with time as the x-axis and uptake as the y-axis. The slope obtained is the initial uptake rate at that calcium ion concentration. The uptake rate was divided by the protein content of the sample to obtain the normalized uptake rate. A SERCA activity curve was plotted with calcium ion concentration as the x-axis and normalized uptake rate as the y-axis. The maximum uptake rate and apparent affinity of calcium ions could be obtained by fitting the curve, thereby achieving quantitative analysis of SERCA calcium pump activity.

[0043] In practical applications, the number of flow channels can be adjusted according to the experimental throughput requirements. In addition to the 14 channels described in Example 1, it can also be set to 6, 8, 12, 16 or 24, etc. Different numbers of flow channels are arranged in an equally spaced array on the experimental cover 2. Increasing the number of channels can improve the batch processing capacity, but the pumping speed of the negative pressure pump needs to be increased accordingly to maintain sufficient negative pressure. Those skilled in the art can select the appropriate number of channels according to the conventional experimental requirements.

[0044] Negative pressure connector 12 is connected to the inlet of the buffer bottle via a pressure-resistant silicone tube, and the outlet of the buffer bottle is connected to the air inlet of the negative pressure pump. The volume of the buffer bottle should be larger than the volume of the storage chamber 11 to prevent liquid from being accidentally sucked into the negative pressure pump. A 0.22-micron air filter can also be installed between the negative pressure pump and the buffer bottle to prevent microorganisms or particulate matter in the air from contaminating the system. Drain connector 13 is connected to the waste liquid collection bottle via a silicone tube. The waste liquid collection bottle should be placed below the negative pressure box 1 so that the waste liquid can be discharged by gravity. The waste liquid collection bottle must have a radioactive marking and a sealed cap, and comply with the isotope waste liquid management regulations.

[0045] It should be understood that the above-described embodiments or examples of the present invention can be combined with each other and have corresponding technical effects.

[0046] The above are merely preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. An experimental apparatus for quantitative analysis of isotope calcium 45 uptake, characterized in that, Includes a negative pressure chamber (1), an experimental cover (2), an assembly filter plate (3), a control valve (4), and an experimental filter membrane (5); The negative pressure box (1) has a storage cavity (11) on its inner side, an open top on its top, and a negative pressure connector (12) and a drain connector (13) communicating with the storage cavity (11) on its side wall. The experimental cover (2) is fixedly installed at the opening on the top of the negative pressure box (1), and the connection between the experimental cover (2) and the negative pressure box (1) is sealed; a through flow groove is provided on the experimental cover (2), and the flow groove is connected to the storage cavity (11); The control valve (4) is installed on the experimental cover (2) and cooperates with the flow channel. The control valve (4) can control the opening and closing of the flow channel. The assembled filter plate (3) is disposed in the flow groove, and the connection between the assembled filter plate (3) and the flow groove is sealed. The experimental filter membrane (5) is laid flat on the upper surface of the assembled filter plate (3).

2. The experimental apparatus for quantitative analysis of calcium-45 isotope uptake according to claim 1, characterized in that, The assembly filter plate (3) is set at a height lower than the horizontal height of the upper surface of the experimental cover (2), so that the assembly filter plate (3) and the flow channel form a groove that can store experimental materials.

3. The isotope calcium 45 uptake quantification assay apparatus according to claim 1, wherein, The flow channel is provided in multiple ways, and the multiple flow channel array is arranged on the experimental cover (2); each flow channel is provided with an assembly filter plate (3), a control valve (4) and an experimental filter membrane (5), and the opening and closing of a single flow channel can be controlled individually through the corresponding control valve (4).

4. The isotope calcium 45 uptake quantification assay apparatus according to claim 1, wherein, The experimental cover (2) is provided with a horizontal limiting groove (21), which can pass through the flow groove; the control valve (4) includes a cylindrical valve body (41) and an adjusting head (42), a valve port (411) is provided through the valve body (41), the valve body (41) is rotatably embedded in the limiting groove (21), and the valve port (411) is located at the flow groove; rotating the valve body (41) can adjust the communication state between the valve port (411) and the flow groove; the adjusting head (42) is located on the outside of the experimental cover (2), and rotating the adjusting head (42) can drive the valve body (41) to rotate.

5. The isotope calcium 45 uptake quantification assay device according to claim 4, characterized in that, The adjusting head (42) is provided with an identification line (421), which is used to indicate the opening direction of the valve port (411).

6. The isotope calcium 45 uptake quantification assay apparatus according to claim 1, wherein, The assembled filter plate (3) is a quartz sand filter plate.

7. The isotope calcium 45 uptake quantification assay apparatus according to claim 1, wherein, It also includes a radiation baffle (6), which is mounted on the side of the negative pressure box (1); the upper part of the radiation baffle (6) is bent to form an observation section.

8. The isotope calcium 45 uptake quantification assay apparatus according to claim 1, wherein, The negative pressure connector (12) is located on the upper side wall of the negative pressure box (1) to prevent the liquid in the storage chamber (11) from being discharged through the negative pressure connector (12); the drain connector (13) is located at the bottom side wall of the negative pressure box (1) to drain the liquid in the storage chamber (11).

9. An experimental method for quantitative analysis of isotope calcium 45 uptake, characterized by, Using the quantitative analysis apparatus for isotope calcium-45 uptake according to any one of claims 1 to 8, the method comprises the following steps: S1. After wetting the experimental filter membrane (5), lay it flat on the upper surface of the assembled filter plate (3) placed in the flow channel, and close the control valve (4). S2. Incubate the reaction solution containing endoplasmic reticulum vesicles and calcium 45 isotope at the set calcium ion concentration and time. S3. After the incubation is completed, the reaction solution is transferred to the filter membrane on the upper surface of the filter plate in the flow channel. The control valve (4) is opened and the negative pressure is started so that the reaction solution enters the storage chamber (11) after being filtered through the experimental filter membrane (5). The endoplasmic reticulum vesicles are trapped on the experimental filter membrane (5). S4. Close the control valve (4) and repeat steps S1 to S3 to process other samples in batches. S5. After the experimental filter membrane (5) is dried, it is transferred to the scintillation tube, scintillation fluid is added, and isotope concentration is detected.

10. The method for quantitative analysis of the isotope calcium 45 uptake according to claim 9, characterized in that, In step S3, the reaction solution is transferred in its entirety to the flow channel for filtration. In step S5, the experimental filter membrane (5) is dried and then transferred as a whole to the scintillation tube.

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