Density gradient centrifugation sample separation and collection device

Abstract

The invention discloses a density gradient centrifugation sample separation and collection device which comprises a body, a separation analysis part, a recovery part and a main control part. Wherein a sample groove is formed in the body; the separation analysis part is arranged at the top of the body, a sample channel is formed in the separation analysis part, and the inlet end of the sample channel corresponds to the sample groove; the recovery part is arranged at the top of the body and corresponds to the outlet end of the sample channel; the main control part is arranged on the body, controls the sample groove to move towards the separation analysis part so as to enable the density gradient centrifugal samples of different layers to sequentially enter the sample channel for separation, and controls the recovery part to respectively receive the separated density gradient centrifugal samples. Work is controlled by the main control part, so that a density gradient centrifugal sample is separated after quickly passing through the separation analysis part, the operation is simple, the flow time is short, the sample separation accuracy can be ensured to the greatest extent, the pollution in the whole separation process is reduced, and the separation and storage of a biological sample are facilitated.

Description

Technical Field

[0001] The invention relates to the technical field of separation and collection of products after centrifugation, in particular to a density gradient centrifugation sample separation and collection device. Background Art

[0002] Ultracentrifugation is the most commonly used technique for separating and purifying biomacromolecules. When centrifuging different particles with small differences, density gradient centrifugation is often relied on. Density gradient centrifugation can separate samples with similar sedimentation coefficients. The samples are separated by centrifugal force in the density gradient medium and finally distributed to specific positions in the gradient medium. Density gradient centrifugation is a commonly used method for separating biomolecules. It is widely used in the fields of virus separation and purification, protein separation, etc. It generally includes three steps: first, prepare the density gradient medium in the centrifuge tube, and the sample to be separated is above the medium; use an ultracentrifuge to centrifuge so that the components in the sample to be separated are layered in the centrifuge tube; finally, collect and detect the distribution of samples in each gradient in the medium after centrifugation. Therefore, the final key to the density gradient method is how to take out the layered samples after centrifugation layer by layer without causing disturbance of the upper and lower layers in the gradient medium, so as to ensure the purity of each layer of samples after separation.

[0003] At present, the collection and detection of the final sample of the density gradient method mainly uses a pipette or a syringe to extract the centrifuged sample by volume and then analyze it. However, this method has a large disturbance to the sample, and there is blindness in sample collection, which is time-consuming and requires high manual operation, and it is difficult to meet the user's demand for rapid sample separation. Therefore, the prior art began to use a fully automatic density gradient separation system to achieve fully automated sample separation, detection, and collection. However, this type of fully automatic density gradient separation system still has the problems of cumbersome operation, high requirements for the operating environment, and is not conducive to the preservation of biological samples.

[0004] Therefore, the prior art still needs to be improved and developed. Summary of the invention

[0005] In view of the above-mentioned deficiencies in the prior art, the purpose of the present invention is to provide a density gradient centrifugation sample separation and collection device, aiming to solve the problem in the prior art that the separation process of density gradient centrifugation separation samples is complicated, the steps are cumbersome and the operating environment requirements are high, which is not conducive to the separation and preservation of biological samples.

[0006] The present invention provides a density gradient centrifugation sample separation and collection device, which comprises:

[0007] A body, wherein a sample slot is provided inside the body for placing density gradient centrifugation samples;

[0008] A separation and analysis part, wherein the separation and analysis part is arranged at the top of the body, a sample channel is formed inside the separation and analysis part, and an inlet end of the sample channel is arranged corresponding to the sample slot to receive a density gradient centrifugation sample;

[0009] A recovery part, which is arranged at the top of the body and corresponds to the outlet end of the sample channel to receive the density gradient centrifugation sample passing through the sample channel;

[0010] The main control unit is arranged on the main body, controls the sample slot to move toward the separation and analysis unit so that the density gradient centrifugation samples of different layers enter the sample channel in sequence for separation, and controls the recovery unit to receive the separated density gradient centrifugation samples respectively.

[0011] In one embodiment, the sample tank comprises:

[0012] A placement position, which is arranged at the center of the sample slot and is used to place a centrifuge tube containing a density gradient centrifugation sample, and an opening is arranged at the top of the body corresponding to the placement position;

[0013] A first motor is arranged below the placement position, and under the control of the main control unit, pushes the placement position to move in a vertical direction, so that the centrifuge tube in the placement position passes through the opening and enters the separation and analysis part, and the density gradient centrifugation samples of different layers in the centrifuge tube are squeezed into the sample channel in sequence.

[0014] In one embodiment, in the separation and analysis section, the inlet end of the sample channel is in the shape of an inverted trumpet, and a gap is formed between the inlet end and the side wall of the separation and analysis section, so that after the centrifuge tube enters the separation and analysis section, the tube wall of the centrifuge tube is located between the inlet end and the side wall and is sleeved on the inlet end, so that density gradient centrifugation samples of different layers in the centrifuge tube are squeezed into the sample channel in sequence.

[0015] In one embodiment, the separation and analysis unit further comprises:

[0016] A sealing ring, which is arranged outside the inlet end of the sample channel to seal the centrifuge tube and the inlet end of the sample channel after the centrifuge tube is sleeved on the inlet end, so as to ensure that the density gradient centrifugation sample enters the sample channel in a layered order;

[0017] The analytical detector is arranged on both sides of the sample channel to obtain data of the density gradient centrifugation sample passing through the sample channel to determine the time when different layers of density gradient centrifugation samples pass through the sample channel to achieve separation of the density gradient centrifugation samples.

[0018] In one embodiment, the recovery unit comprises:

[0019] A test tube rack, wherein the test tube rack is provided with a plurality of test tube positions for placing a plurality of test tubes;

[0020] A second motor is disposed below the test tube rack, and controls the test tube rack to move so that the test tubes correspond to the outlet ends of the sample channels in sequence, so as to respectively receive the separated density gradient centrifugation samples.

[0021] In one embodiment, the test tube rack is a disc structure, and the second motor controls the test tube rack to rotate so that the plurality of test tubes correspond to the outlet ends of the sample channels in sequence.

[0022] In one embodiment, the main control unit includes:

[0023] A main control panel, which is disposed on a side of the body and receives the acquired density gradient centrifugation sample data from the analytical detector and generates a waveform diagram to determine the composition of the density gradient centrifugation sample currently flowing through the sample channel;

[0024] A main control button is arranged below the main control panel to control the operation of the density gradient centrifugation sample separation device and set corresponding working parameters.

[0025] In one embodiment, it further includes:

[0026] A fixed bracket is fixedly arranged on the top of the main body and is used to clamp and fix the separation and analysis part so that the inlet end of the sample channel is fixed above the opening.

[0027] In one embodiment, it further includes:

[0028] An ultrasonic cleaning unit, which is disposed on the top of the body and is used to clean the separation and analysis unit after removing the separation and analysis unit from the fixed support;

[0029] A temperature control unit is disposed around the body to control the temperature of the density gradient centrifugation sample in the density gradient centrifugation sample separation and collection device.

[0030] In one embodiment, a sliding door is provided on the side of the body to place a centrifuge tube containing a density gradient centrifugation sample into the sample slot.

[0031] In summary, the present invention discloses a density gradient centrifugal sample separation and collection device, comprising: a body, a separation and analysis unit, a recovery unit and a main control unit. Among them, a sample slot is provided inside the body to place the density gradient centrifugal sample; the separation and analysis unit is arranged at the top of the body, and a sample channel is formed inside the separation and analysis unit, and the inlet end of the sample channel is arranged corresponding to the sample slot to receive the density gradient centrifugal sample; the recovery unit is arranged at the top of the body and corresponds to the outlet end of the sample channel to receive the density gradient centrifugal sample passing through the sample channel; the main control unit is arranged on the body, controls the sample slot to move toward the separation and analysis unit so that the density gradient centrifugal samples of different layers enter the sample channel in sequence for separation, and controls the recovery unit to receive the separated density gradient centrifugal samples respectively. The present invention controls the work through the main control unit, so that the density gradient centrifugal sample can be separated after passing through the separation and analysis unit quickly, the operation is simple, the process time is short, the accuracy of sample separation can be ensured to the greatest extent, and the pollution of the entire separation process can be reduced, which is conducive to the separation and preservation of biological samples. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings in the following description are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0033] Figure 1 This is a schematic diagram of the structural explosion of the density gradient centrifugation sample separation and collection device of the present invention.

[0034] Figure 2 It is a schematic diagram of an explosion after the separation and analysis part is clamped on the fixed bracket in the density gradient centrifugation sample separation and collection device of the present invention.

[0035] Figure 3 It is a side view of the density gradient centrifugation sample separation and collection device of the present invention.

[0036] Figure 4 It is a side sectional view of the density gradient centrifugation sample separation and collection device of the present invention.

[0037] Figure 5 This is a schematic diagram of the structural explosion of another embodiment of the density gradient centrifugation sample separation and collection device of the present invention.

[0038] Figure 6 It is a side view of another embodiment of the density gradient centrifugation sample separation and collection device of the present invention.

[0039] Figure 7 It is a schematic diagram of the corresponding relationship between the centrifuge tube and the separation and analysis part in the density gradient centrifugation sample separation and collection device of the present invention.

[0040] Figure 8 This is a spectral image detected by the separation and analysis part of the density gradient centrifugation sample separation and collection device in one embodiment of the present invention. DETAILED DESCRIPTION

[0041] The present invention provides a density gradient centrifugal sample separation and collection device. To make the purpose, technical solution and effect of the present invention clearer and more specific, the present invention is further described in detail below. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0042] It should be noted that the directions or positional relationships indicated by terms such as “center”, “up”, “down”, “left”, “right”, “inside”, “outside”, “vertical” and “horizontal” are based on the directions or positional relationships shown in the accompanying drawings and are only for the convenience of describing the present invention and simplifying the description. They do not indicate or imply that the structure referred to must have a specific direction or must be constructed in a specific direction, and should not be understood as a limitation on the present invention.

[0043] In addition, unless there is a special limitation on the articles in the text, "one" and "the" may refer to a single one or multiple ones. If there are descriptions involving "first", "second", etc. in the embodiments of the present invention, the descriptions of "first", "second", etc. are only used for descriptive purposes and cannot be understood as indicating or implying their relative importance or implicitly indicating the number of technical features indicated. Therefore, the features defined as "first" and "second" may explicitly or implicitly include at least one of the features. In addition, the technical solutions between the various embodiments can be combined with each other, but they must be based on the ability of ordinary technicians in the field to implement them. When the combination of technical solutions is contradictory or cannot be implemented, it should be deemed that such a combination of technical solutions does not exist and is not within the scope of protection required by the present invention.

[0044] Density gradient centrifugation is a common method for separating biomolecules. It is widely used in virus separation and purification, protein separation and other fields. It generally includes three steps: first, prepare a density gradient medium in a centrifuge tube, with the sample to be separated on top of the medium; use an ultracentrifuge to centrifuge so that the components in the sample to be separated are separated into layers in the centrifuge tube; finally, collect and detect the distribution of samples in each gradient in the medium after centrifugation. After the medium is separated into layers by density gradient centrifugation, the components in different layers are generally obtained by cutting, puncture and siphoning. Among them, the cutting method is to put the centrifuge tube and the sample after centrifugation into a refrigerator for quick freezing, and then use a centrifuge tube cutter to cut the frozen centrifuge tube into thin slices, and take out the samples layer by layer, but this method is costly, time-consuming, and not suitable for thick-walled tubes or metal tubes; the puncture method uses a needle to puncture the bottom of the centrifuge tube, allowing the sample to drip freely by gravity, and then distribute and collect in sequence, but this method is also costly and not suitable for the situation where there is precipitation at the bottom after centrifugation; the siphon method is to extend a polyethylene tube from the opening of the centrifuge tube until it is inserted into the bottom of the tube, and use the siphon method to suck out the gradient density centrifugation sample in layers, but this method easily causes the separation zone to be disturbed, affecting the final separation result. Therefore, the separation methods of density gradient centrifugation samples in the prior art are all limited, and it is difficult to ensure fast and accurate sample separation results.

[0045] In order to solve this problem and realize the rapid and accurate separation function of density gradient centrifugation samples, the present invention provides a density gradient centrifugation sample separation and collection device. Specifically, Figure 1 and Figure 2 As shown, the density gradient centrifugal sample separation and collection device comprises: a body 100, a separation and analysis unit 200, a recovery unit 300 and a main control unit 400. Figure 1 and Figure 4As shown, a sample slot 110 is provided inside the body 100 to place a density gradient centrifugation sample; the separation and analysis part 200 is arranged at the top of the body 100, and a sample channel 210 is formed inside the separation and analysis part 200, and an inlet end 211 of the sample channel 210 is arranged corresponding to the sample slot 110 to receive the density gradient centrifugation sample; the recovery part 300 is arranged at the top of the body 100 and is arranged corresponding to the outlet end 212 of the sample channel 210 to receive the density gradient centrifugation sample passing through the sample channel 210; the main control part 400 is arranged on the body 100, controls the sample slot 110 to move toward the separation and analysis part 200 so that different layers of density gradient centrifugation samples enter the sample channel 210 in sequence for separation, and controls the recovery part 300 to receive the separated density gradient centrifugation samples respectively. The density gradient centrifugation sample separation device is controlled by the main control unit 400, so that the density gradient centrifugation sample is quickly separated after passing through the separation and analysis unit 200. The operation is simple and the process time is short. The accuracy of sample separation can be ensured to the greatest extent, and the contamination of the entire separation process can be reduced, which is beneficial to the separation and preservation of biological samples.

[0046] Furthermore, if Figure 1 and Figure 2 As shown, the sample slot 110 includes a placement position 111 and a first motor 112. The placement position 111 is set at the center of the sample slot 110, and is used to place a centrifuge tube containing a density gradient centrifugation sample. The top of the body 100 is provided with an opening 120 corresponding to the placement position 111; the first motor 112 is set below the placement position 111, and under the control of the main control unit 400, the placement position 111 is pushed to move in the vertical direction, so that the centrifuge tube in the placement position 111 passes through the opening 120 and enters the separation and analysis unit 200, and the density gradient centrifugation samples of different layers in the centrifuge tube are squeezed into the sample channel in sequence. The first motor 112 pushes the centrifuge tube to move synchronously with the placement position 111 in the vertical direction, and the layered samples in the centrifuge tube are squeezed into the separation and analysis unit 200 in order from top to bottom, thereby ensuring the order of the samples of each layer entering the separation and analysis unit 200, and at the same time ensuring that there is no contamination between the samples of each layer, so as to achieve accurate and rapid sample separation.

[0047] In one embodiment, Figure 5 and Figure 6 As shown, a sliding door 140 is provided on the side of the body 100 corresponding to the position of the sample slot 110 . The sliding door 140 can be opened and closed freely to facilitate the user to place a centrifuge tube containing a density gradient centrifugation sample in the sample slot 110 .

[0048] Furthermore, if Figure 4 and Figure 7 As shown, the sample channel 210 in the separation and analysis section 200 is an overall slender tubular structure, and a structure with a size gradient change is formed at the inlet end 211 to match the centrifuge tube containing the density gradient centrifugation sample at the opening of the inlet end 211, ensuring that the density gradient centrifugation sample in the centrifuge tube can enter the sample channel 210 in a layered order. Specifically, the sample channel 210 is a capillary structure as a whole, and the inlet end 211 is an inverted trumpet-shaped structure facing the sample slot 110, and a gap is formed between the inlet end 211 and the side wall of the separation and analysis part 200, wherein the outer diameter of the opening of the inlet end 211 is slightly smaller than the inner diameter of the centrifuge tube, and the inner diameter of the side wall of the separation and analysis part 200 is slightly larger than the outer diameter of the centrifuge tube, so that after the centrifuge tube enters the separation and analysis part 200, the tube wall of the centrifuge tube is arranged between the inlet end 211 and the side wall corresponding to the gap, and the centrifuge tube is sleeved on the inlet end 211, thereby ensuring that when the centrifuge tube continues to move toward the sample channel 210, the density gradient centrifugation sample in the centrifuge tube enters the sample channel 210 layer by layer in a top-down order, thereby realizing stable separation between samples in different layers.

[0049] Furthermore, if Figure 4 As shown, the separation and analysis part 200 also includes a sealing ring 220 and an analysis detector 230. The sealing ring 220 is arranged outside the inlet end 211 of the sample channel 210, so as to be sandwiched between the inner wall of the centrifuge tube and the outer wall of the inlet end 211 after the centrifuge tube enters the separation and analysis part 200, thereby sealing the centrifuge tube and the inlet end 211 of the sample channel 210, ensuring that the density gradient centrifugation sample will not leak, and at the same time ensuring that different layers of samples can enter the sample channel 210 in sequence, thereby achieving stable separation between different layers of samples. The analysis detector 230 is arranged on both sides of the sample channel 210, and obtains data of the density gradient centrifugation sample passing through the sample channel 210, so as to determine the time when the density gradient centrifugation samples of different layers pass through the sample channel 210, thereby achieving separation of the density gradient centrifugation samples. The sealing ring 220 seals the inlet end 211 of the centrifuge tube and the sample channel 210, thereby enhancing the sealing between the centrifuge tube and the sample channel 210, improving the accuracy of sample separation, and avoiding external contamination of the sample, thereby achieving separation and preservation of biological samples.

[0050] Furthermore, the analytical detector is a UV detector, which detects the UV spectrum data of the density gradient centrifugation sample passing through the sample channel 210, and records the corresponding passing time, so as to achieve the separation of density gradient centrifugation samples of different components. By forming a UV spectrum to identify the components of the sample, even if there are impurity layer components, or adjacent layer components are mutually soluble, they can be quickly identified and distinguished, and the impurity part and the mutually soluble part can be collected as separate components, thereby ensuring the purity of the collected components of each layer and improving the accuracy of separation. At the same time, through rapid UV detection, the sample does not need to stay in the sample channel 210 for a long time, and the sample channel 210 can be set to a particularly short flow path, which can accurately determine the composition of the sample, thereby reducing contamination between samples of different layers and achieving rapid and accurate sample separation.

[0051] Furthermore, if Figure 1 and Figure 2 As shown, the recovery unit 300 includes a test tube rack 310 and a second motor 320. The test tube rack 310 is provided with a plurality of test tube positions 311 for placing a plurality of test tubes; the second motor 320 is disposed below the test tube rack 310 to control the movement of the test tube rack 310 so that the plurality of test tubes correspond to the outlet end 212 of the sample channel 210 in sequence to receive the separated density gradient centrifugation samples respectively.

[0052] Furthermore, the test tube rack 310 is a disc structure, and the second motor 320 controls the test tube rack 310 to rotate so that the plurality of test tubes correspond to the outlet ends 212 of the sample channels 210 in sequence.

[0053] Furthermore, if Figure 1 and Figure 2 As shown, the main control unit 400 includes a main control panel 410 and a main control button 420. The main control panel 410 is arranged on the side of the body 100, receives the acquired density gradient centrifugation sample data from the analysis detector 230, and generates a waveform diagram to determine the composition of the density gradient centrifugation sample currently flowing through the sample channel 210; the main control button 420 is arranged below the main control panel 410 to control the operation of the density gradient centrifugation sample separation device and set corresponding working parameters. Specifically, the working parameters include the moving speed of the placement position 111 driven by the first motor 112, the rotation speed of the test tube rack 310 driven by the second motor 320, etc., to ensure that the density gradient centrifugation sample can be separated automatically and accurately.

[0054] Furthermore, if Figure 2 and Figure 3As shown, the density gradient centrifugal sample separation and collection device further includes a fixing bracket 500, which is fixedly disposed on the top of the body 100 and is used to clamp and fix the separation and analysis part 200 so that the inlet end 211 of the sample channel 210 is fixed above the opening 120. The fixing bracket 500 realizes a detachable connection between the separation and analysis part 200 and the body 100, thereby facilitating adjustment of the height of the separation and analysis part 200 relative to the opening 120, and cleaning and replacement of the separation and analysis part 200 after removal.

[0055] Furthermore, if Figure 1 and Figure 2 As shown, the density gradient centrifugal sample separation and collection device also includes an ultrasonic cleaning unit 600, which is arranged at the top of the body 100 and is used to remove the separation and analysis unit 200 from the fixed bracket 500 and then clean the separation and analysis unit 200. The ultrasonic cleaning unit 600 is used to clean the separation and analysis unit 200, thereby extending the service life of the density gradient centrifugal sample separation and collection device. Furthermore, the ultrasonic cleaning unit 600 is also provided with a drying device to quickly dry the separation and analysis unit 200 after ultrasonic cleaning, so as to facilitate the next use.

[0056] Furthermore, if Figure 4 As shown, the density gradient centrifugal sample separation and collection device also includes a temperature control unit 700, which is arranged around the body 100 to control the temperature of the density gradient centrifugal sample in the density gradient centrifugal sample separation and collection device. Optionally, the temperature control unit 700 is set outside the body 100 through a structure similar to a protective cover, so as to control the temperature of the entire density gradient centrifugal sample separation and collection device, thereby ensuring that the temperature of the density gradient centrifugal sample to be separated, the density gradient centrifugal sample in separation, and the separated density gradient centrifugal sample are all kept within a suitable range. The temperature of the density gradient centrifugal sample in the density gradient centrifugal sample separation and collection device of the present invention is adjusted by the temperature control unit 700 to ensure that the sample is at the optimal temperature, thereby avoiding sample inactivation, improving sample stability, and realizing separation and collection of biological samples. Optionally, the temperature control unit 700 is an air conditioning circulation system, which is respectively arranged at different positions of the density gradient centrifugal sample separation and collection device, and controls the temperature of the density gradient centrifugal sample in the density gradient centrifugal sample separation and collection device through compressor refrigeration.

[0057] The present invention controls the operation of the density gradient centrifugation sample separation device through a main control unit, so that the density gradient centrifugation sample is quickly separated after passing through a separation and analysis unit. The operation is simple and the process time is short. The accuracy of sample separation can be ensured to the greatest extent, and contamination in the entire separation process can be reduced, which is beneficial to the separation and preservation of biological samples.

[0058] In one embodiment, Figure 1 and Figure 4 As shown, the density gradient centrifugal sample separation and collection device of the present invention is composed of a body 100, a separation and analysis unit 200, a recovery unit 300, a main control unit 400, a fixed bracket 500, an ultrasonic cleaning unit 600 and a temperature control unit 700. In one embodiment, the temperature control unit 700 forms a protective cover structure and is arranged outside the body 100. The temperature control unit 700 is arranged on the body 100 from above and accommodates the body 100, the separation and analysis unit 200, the recovery unit 300, the main control unit 400, the fixed bracket 500 and the ultrasonic cleaning unit 600, so as to avoid the influence of the external environment on the various components in the density gradient centrifugal sample separation and collection device, and control the temperature in the density gradient centrifugal sample separation and collection device as needed.

[0059] Specifically, Figure 1 As shown, in this embodiment, the body 100 is a hollow box structure, wherein the body top plate 130 of the body 100 can be removed to expose the sample tank 110 arranged inside the body 100. Specifically, the sample tank 110 is a circular tank, and the center position is depressed downward to form a placement position 111, which is used to place centrifuge tubes that have been centrifuged and have been layered by density gradient inside. The sample tank 110 is accommodated inside the body 100 to avoid contamination and extend the storage time of density gradient centrifugation components in the sample tank 110. Further, a first motor 112 is provided below the placement position 111, and the first motor 112 can be a stepping motor to control the placement position 111 to move in the vertical direction. An opening 120 is provided at the position of the body top plate 130 corresponding to the placement position 111, so that when the first motor 112 drives the placement position 111 to move upward, the centrifuge tube placed in the placement position 111 can extend from the body top plate 130.

[0060] Further, in another embodiment, Figure 5 and Figure 6 As shown, a sliding door 140 is provided on the side of the main body 100. When the sliding door 140 is opened, a centrifuge tube containing a density gradient centrifugation sample can be placed in the sample slot 110. When the sliding door 140 is closed, the centrifuge tube placed in the sample slot 110 is sealed in the main body 100.

[0061] Furthermore, in another embodiment, the top plate of the main body 100 is fixed, and a drawer structure is formed on the side of the main body 100. The sample slot 110 is fixed on the drawer structure. The sample slot 110 is taken out of the main body 100 by pulling the drawer structure, so that the centrifuge tube to be processed can be placed on the placement position 111.

[0062] Furthermore, if Figure 2 and Figure 3 As shown, in this embodiment, the main control unit 400 is arranged on the side of the body 100, and the main control unit 400 includes a main control panel 410 and a main control button 420, wherein the main control panel 410 is used to display the data of the density gradient centrifugation sample obtained by the separation and analysis unit 200, so that the user can determine the sample components currently flowing out of the separation and analysis unit 200, and the main control button 420 is respectively connected to the sample tank 110, the separation and analysis unit 200, the recovery unit 300, the ultrasonic cleaning unit 600 and the temperature control unit 700 for communication, so as to control the working parameters of each component in the density gradient centrifugation sample separation and collection device. Optionally, the working parameters include parameters such as injection speed, recovery conditions, temperature conditions and ultrasonic parameters, so as to realize the automatic processing of the density gradient centrifugation sample separation and collection device.

[0063] Furthermore, if Figure 1 As shown, in this embodiment, the temperature control unit 700 is arranged outside the body 100 through a protective cover structure, so that the temperature control unit 700 is arranged around the body 100. Optionally, the temperature control unit 700 is an air conditioner arranged around the body, and the air conditioner is connected to the main control unit 400 for communication, and the power of the air conditioner is controlled by the main control unit 400, thereby controlling the temperature of the density gradient centrifugation sample to be separated, the density gradient centrifugation sample in the separation process, and the separated density gradient centrifugation sample in the density gradient centrifugation sample separation and collection device, ensuring that the corresponding samples are stored at a suitable temperature.

[0064] Furthermore, if Figure 1 and Figure 2As shown, in this embodiment, a fixed bracket 500 is fixedly provided on the main body top plate 130 at the top of the main body 100, and the fixed bracket 500 is clamped and connected with the separation and analysis part 200, so as to realize the detachable connection between the separation and analysis part 200 and the main body 100, and when the fixed bracket 500 is clamped with the separation and analysis part 200, the separation and analysis part 200 is located above the opening 120, so as to ensure that when the first motor 112 pushes the placement position 111 to move upward, the centrifuge tube in the placement position 111 can be directly extended into the separation and analysis part 200. Optionally, the fixed bracket 500 is a bracket structure with adjustable height to adjust the height of the separation and analysis part 200 relative to the opening 120, so as to match centrifuge tubes of different sizes placed in the placement position 111.

[0065] Furthermore, if Figure 1 and Figure 2 As shown, in this embodiment, an ultrasonic cleaning unit 600 is provided on the main body top plate 130, and the ultrasonic cleaning unit 600 is an ultrasonic cleaning tank formed on the top surface of the main body top plate 130. After the separation and analysis unit 200 is used, the separation and analysis unit 200 is removed from the fixing bracket 500 and placed in the ultrasonic cleaning unit 600, and the main control unit 400 controls the ultrasonic cleaning unit 600 to start working, thereby cleaning the separation and analysis unit 200 and preventing residual components in the separation and analysis unit 200 from affecting the next separation result.

[0066] Furthermore, if Figure 4As shown, in this embodiment, a sample channel 210 is formed in the separation and analysis part 200 for density gradient centrifugation samples to pass through, and the density gradient centrifugation samples passing through the sample channel 210 are separated. Specifically, the sample channel 210 is a capillary channel, including an inlet end 211 and an outlet end 212 in the vertical direction, and a connecting tube 213 connecting the inlet end 211 and the outlet end 212 in the horizontal direction. Among them, the inlet end 211 is arranged above the opening 120 corresponding to the centrifuge tube arrangement in the placement position 111, and the inlet end 211 is arranged in an inverted trumpet shape and forms a gap with the side wall of the separation and analysis part 200, so that when the first motor 112 drives the placement position 111 to move upward, the tube wall of the centrifuge tube placed in the placement position 111 is stuck in the gap, so that the centrifuge tube is sleeved on the inlet end 211. Furthermore, an elastic sealing ring 220 is provided on the outside of the inlet end 211 to seal the gap between the centrifuge tube and the inlet end 211 after the centrifuge tube enters the separation and analysis part 200 and is sleeved on the inlet end 211, so that when the first motor 112 drives the placement position 111 to continue to rise, the centrifuge tube is squeezed to completely squeeze the density gradient centrifugation components inside into the inlet end 211 without leaking to a position outside the sample channel. Since the sample channel 210 is a capillary channel, the density gradient centrifugation components enter the sample channel in sequence from top to bottom and move along the sample channel 210, and finally drip out from the outlet end 212 in sequence. Furthermore, the sealing ring 220 ensures the sealing of the centrifuge tube and the inlet end 211, prevents air or other impurities from entering the sample channel 210, and thus improves the accuracy of sample separation.

[0067] Furthermore, if Figure 4As shown, in this embodiment, an analytical detector 230 is provided on the side of the connecting tube 213 of the sample channel 210. The analytical detector 230 is a UV detector, which determines the composition of the current sample component by detecting the UV spectrum data of the sample component flowing through the specific position of the sample channel 210. Further, the analytical detector 230 sends the acquired UV spectrum data to the main control unit 400, and generates a spectrum diagram on the main control panel 410 to indicate the composition of the current sample component. When the UV spectrum data detected by the analytical detector 230 changes, it means that the components of the same layer in the original density gradient centrifugation component have all flowed through the sample channel 210, and the components that continue to flow through the sample channel 210 are the components of the next layer. The time when the component composition changes is recorded, so as to determine the time when different components drip out from the outlet 212, so as to achieve the separation of different layers of components in the density gradient centrifugation component. Optionally, the density gradient medium in the density gradient centrifugation component is a sucrose solution. Furthermore, the composition of the current component is determined by UV spectral data. In this case, even if there is mutual solubility between adjacent layers of the sample to be separated, they can be identified and distinguished through different spectral waveforms on the main control panel 410, and the mutual solubility parts can be collected as separate components, thereby ensuring the purity of the collected components in each layer, avoiding mutual interference between components in adjacent layers, and improving the accuracy of separation.

[0068] Furthermore, if Figure 1 and Figure 2As shown, in this embodiment, the recovery part 300 is arranged on the top surface of the main body top plate 130 and is arranged corresponding to the outlet end 212 of the sample channel 210 in the separation and analysis part 200. Specifically, the recovery part 300 includes a disc-shaped test tube rack 310 and a second motor 320 arranged below the test tube rack 310. Among them, the test tube rack 310 is evenly spaced along the circumferential direction to place multiple test tubes to receive density gradient centrifugation components of different components. Specifically, after the main control unit 400 obtains the UV spectrum data and corresponding time data collected by the analysis detector 230 in the separation and analysis unit 200, the start time and end time of the components of different layers in the density gradient centrifugation components dripping out from the sample channel 210 are determined, and the main control unit 400 controls the second motor 320 to drive the test tube rack 310 to rotate, so as to ensure that during the time period when the components of the same layer drip out from the sample channel 210, the outlet end 212 of the sample channel 210 corresponding to the same test tube receives the corresponding density gradient centrifugation components; and after all the components of the same layer drip out, the test tube corresponding to the outlet end 212 of the sample channel 210 is switched, so as to ensure that the density gradient centrifugation components of different layers are received by different test tubes, and the separation and collection of the density gradient centrifugation components are completed. At the same time, combined with the spectrum generated on the main control panel 410, the components of the density gradient centrifugation components in each test tube can be determined, so as to achieve rapid and accurate separation, collection and analysis of the density gradient centrifugation components.

[0069] like Figure 8 The figure shows that when the density gradient centrifugation sample separation and collection device of the present invention is used to separate the ribosome components of yeast cells, the corresponding components flow through the sample channel 210 and sequentially pass through the analysis detector 230, and the analysis detector 230 detects the acquired spectral data. Specifically, in this embodiment, the preparation process of the density gradient centrifugation sample to be separated is: first, a 10% sucrose solution and a 60% sucrose solution are slowly added from the bottom of the SW40 sample tube using a syringe, and the solution is left to stand overnight to obtain a uniformly distributed density gradient centrifugation solution; then, the yeast cells are centrifuged and quickly ground with liquid nitrogen, and centrifuged at 14000rpm and 4℃ for 20min, at which time the supernatant contains a large number of ribosomes; then, the supernatant is slowly added to the top of the sucrose density gradient solution, and an ultracentrifuge is used to centrifuge at 38krpm and 4℃ for 2h40min to obtain a density gradient centrifugation sample, at which time different components of the ribosome are distributed in samples of different densities. The density gradient centrifugation sample in this embodiment is placed in the sample slot 110 of the density gradient centrifugation sample separation and collection device of the present invention, the temperature is set to 4°C by the temperature control unit 700, and the centrifuge tube is injected into the separation and analysis unit 200 at 1 mL / min to obtain the following: Figure 8The peak graph shown. According to the peak data in the image, the solutions passing through the separation detector 230 in the separation and analysis unit 200 are the solution containing the 40S ribosome small subunit, the solution containing the 60S ribosome large subunit, and the solution containing the 80S monoribosome, followed by the solution containing the polysome. According to the corresponding sequence, the main control unit 400 controls the second motor 320 to drive the test tube rack 310 to rotate, so that different test tubes are set corresponding to the outlet end 212 of the sample channel 210 in the corresponding time period to collect the corresponding components respectively, thereby completing the rapid and accurate separation, collection and analysis of different yeast cell ribosome components. The overall process is convenient and fast, and easy to use.

[0070] In summary, the present invention discloses a density gradient centrifugal sample separation and collection device, comprising: a body, a separation and analysis unit, a recovery unit and a main control unit. Among them, a sample slot is provided inside the body to place the density gradient centrifugal sample; the separation and analysis unit is arranged at the top of the body, and a sample channel is formed inside the separation and analysis unit, and the inlet end of the sample channel is arranged corresponding to the sample slot to receive the density gradient centrifugal sample; the recovery unit is arranged at the top of the body and corresponds to the outlet end of the sample channel to receive the density gradient centrifugal sample passing through the sample channel; the main control unit is arranged on the body, controls the sample slot to move toward the separation and analysis unit so that the density gradient centrifugal samples of different layers enter the sample channel in sequence for separation, and controls the recovery unit to receive the separated density gradient centrifugal samples respectively. The present invention controls the work through the main control unit, so that the density gradient centrifugal sample can be separated after passing through the separation and analysis unit quickly, the operation is simple, the process time is short, the accuracy of sample separation can be ensured to the greatest extent, and the pollution of the entire separation process can be reduced, which is conducive to the separation and preservation of biological samples.

[0071] The following is a brief description of the workflow of the density gradient centrifugation sample separation and collection device of the present invention:

[0072] The sliding door 140 is opened, and the centrifuge tube containing the sample to be separated is placed on the placement position 111 of the sample slot 110 in the body 100, and the temperature control unit 700 ensures that the overall environment is at a temperature suitable for each component sample; the working parameters are set at the main control unit 400, and the density gradient centrifugation sample separation and collection device is controlled to work; the first motor 112 pushes the placement position 111 to move upward in the vertical direction, and the centrifuge tube passes through the opening 120 to enter the separation and analysis unit 200, and is sleeved outside the inlet end 211 of the sample channel 210, and the gap between the centrifuge tube and the inlet end 211 is sealed by the sealing ring 220; the first motor 112 continues to push The placement position 111 moves upward in the vertical direction to squeeze the sample to be separated in the centrifuge tube into the sample channel 210; the spectral absorption data of the sample to be separated is collected by the analysis detector 230 in the separation and analysis unit 200, and a spectrum is drawn and displayed on the main control panel 410 to determine the components of different layers of samples in the sample to be separated and the time for each layer of samples to flow through the sample channel 210; the second motor 320 in the recovery unit 300 drives the test tube rack 310 to rotate, so that different components in the sample to be separated are collected separately using different test tubes when they drip out from the outlet end 212 of the sample channel 210, so as to complete the separation, analysis and collection of the components to be separated. After the work is completed, the separation and analysis unit 200 is removed from the fixed bracket 500, placed in the ultrasonic cleaning unit 600 for ultrasonic cleaning and drying, so as to facilitate continued use next time.

[0073] The present invention allows the centrifuge tube containing the sample to be separated to move upward, and continues to squeeze the centrifuge tube after the centrifuge tube is connected to the separation and analysis part, thereby ensuring that the sample to be separated in the centrifuge tube enters the sample channel of the separation and analysis part in a layered order from top to bottom. At the same time, the spectral data of the sample flowing through the sample channel is detected in real time by an analytical detector to determine the components of the current sample, thereby determining the initial time and end time of the components of different components dripping out from the sample channel, so that the components of different components are collected by the recovery part using different test tubes, thereby realizing the rapid separation, analysis and collection of the sample to be separated. Due to the fast detection speed, the sample channel can be set to be very short, which can maximize the accuracy of the separated sample and reduce the contamination between samples. At the same time, since the components of the current component are determined by spectral data, even if there is a mutual solubility between adjacent layers of the sample to be separated, it can be quickly identified in the separation and analysis part of the present invention, and the mutual solubility part is collected as a separate component, thereby ensuring the purity of the components of each layer collected, avoiding mutual interference between components of adjacent layers, further improving the accuracy of separation, and avoiding contamination between the samples finally collected.

[0074] In summary, the present invention discloses a density gradient centrifugal sample separation and collection device, comprising: a body, a separation and analysis unit, a recovery unit and a main control unit. Among them, a sample slot is provided inside the body to place the density gradient centrifugal sample; the separation and analysis unit is arranged at the top of the body, and a sample channel is formed inside the separation and analysis unit, and the inlet end of the sample channel is arranged corresponding to the sample slot to receive the density gradient centrifugal sample; the recovery unit is arranged at the top of the body and corresponds to the outlet end of the sample channel to receive the density gradient centrifugal sample passing through the sample channel; the main control unit is arranged on the body, controls the sample slot to move toward the separation and analysis unit so that the density gradient centrifugal samples of different layers enter the sample channel in sequence for separation, and controls the recovery unit to receive the separated density gradient centrifugal samples respectively. The present invention controls the work through the main control unit, so that the density gradient centrifugal sample can be separated after passing through the separation and analysis unit quickly, the operation is simple, the process time is short, the accuracy of sample separation can be ensured to the greatest extent, and the pollution of the entire separation process can be reduced, which is conducive to the separation and preservation of biological samples.

[0075] The embodiments described above are only used to illustrate the technical solutions of the present invention, rather than to limit the same. Although the present invention has been described in detail with reference to the aforementioned embodiments, it should be understood by those skilled in the art that the technical solutions described in the aforementioned embodiments may still be modified, or some of the technical features therein may be replaced by equivalents. However, these modifications or replacements do not deviate from the spirit and scope of the technical solutions of the embodiments of the present invention, and should be included in the protection scope of the present invention.

Claims

1. A density gradient centrifugation sample separation and collection device, characterized in that: include: A body, wherein a sample slot is provided inside the body for placing density gradient centrifugation samples; A separation and analysis part, wherein the separation and analysis part is arranged at the top of the body, a sample channel is formed inside the separation and analysis part, and an inlet end of the sample channel is arranged corresponding to the sample slot to receive a density gradient centrifugation sample; A recovery part, which is arranged at the top of the body and corresponds to the outlet end of the sample channel to receive the density gradient centrifugation sample passing through the sample channel; The main control unit is arranged on the main body, controls the sample slot to move toward the separation and analysis unit so that the density gradient centrifugation samples of different layers enter the sample channel in sequence for separation, and controls the recovery unit to receive the separated density gradient centrifugation samples respectively.

2. The density gradient centrifugation sample separation and collection device according to claim 1, characterized in that: The sample tank comprises: A placement position, which is arranged at the center of the sample slot and is used to place a centrifuge tube containing a density gradient centrifugation sample, and an opening is arranged at the top of the body corresponding to the placement position; A first motor is arranged below the placement position, and under the control of the main control unit, pushes the placement position to move in a vertical direction, so that the centrifuge tube in the placement position passes through the opening and enters the separation and analysis part, and the density gradient centrifugation samples of different layers in the centrifuge tube are squeezed into the sample channel in sequence.

3. The density gradient centrifugation sample separation and collection device according to claim 2, characterized in that: In the separation and analysis section, the inlet end of the sample channel is in the shape of an inverted trumpet, and a gap is formed between the inlet end and the side wall of the separation and analysis section, so that after the centrifuge tube enters the separation and analysis section, the tube wall of the centrifuge tube is located between the inlet end and the side wall and is sleeved on the inlet end, so that density gradient centrifugation samples of different layers in the centrifuge tube are squeezed into the sample channel in sequence.

4. The density gradient centrifugation sample separation and collection device according to claim 3, characterized in that: The separation and analysis unit further comprises: A sealing ring, which is arranged outside the inlet end of the sample channel to seal the centrifuge tube and the inlet end of the sample channel after the centrifuge tube is sleeved on the inlet end, so as to ensure that the density gradient centrifugation sample enters the sample channel in a layered order; The analytical detector is arranged on both sides of the sample channel to obtain data of the density gradient centrifugation sample passing through the sample channel to determine the time when different layers of density gradient centrifugation samples pass through the sample channel to achieve separation of the density gradient centrifugation samples.

5. The density gradient centrifugation sample separation and collection device according to claim 4, characterized in that: The recovery unit comprises: A test tube rack, wherein the test tube rack is provided with a plurality of test tube positions for placing a plurality of test tubes; A second motor is disposed below the test tube rack, and controls the test tube rack to move so that the test tubes correspond to the outlet ends of the sample channels in sequence, so as to respectively receive the separated density gradient centrifugation samples.

6. The density gradient centrifugation sample separation and collection device according to claim 5, characterized in that: The test tube rack is a disc structure, and the second motor controls the test tube rack to rotate so that the plurality of test tubes correspond to the outlet ends of the sample channels in sequence.

7. The density gradient centrifugation sample separation and collection device according to claim 4, characterized in that: The main control unit comprises: A main control panel, which is disposed on a side of the body and receives the acquired density gradient centrifugation sample data from the analytical detector and generates a waveform diagram to determine the composition of the density gradient centrifugation sample currently flowing through the sample channel; A main control button is arranged below the main control panel to control the operation of the density gradient centrifugation sample separation device and set corresponding working parameters.

8. The density gradient centrifugation sample separation and collection device according to claim 2, characterized in that: Also includes: A fixed bracket is fixedly arranged on the top of the main body and is used to clamp and fix the separation and analysis part so that the inlet end of the sample channel is fixed above the opening.

9. The density gradient centrifugation sample separation and collection device according to claim 8, characterized in that: Also includes: An ultrasonic cleaning unit, which is disposed on the top of the body and is used to clean the separation and analysis unit after removing the separation and analysis unit from the fixed support; A temperature control unit is disposed around the body to control the temperature of the density gradient centrifugation sample in the density gradient centrifugation sample separation and collection device.

10. The density gradient centrifugation sample separation and collection device according to claim 1, characterized in that: A sliding door is provided on the side of the body so as to place a centrifuge tube containing a density gradient centrifugation sample into the sample slot.

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