Combined high-throughput high-speed drop temperature device

Abstract

A novel combined high-flux high-speed cooling and rewarming device includes an upper cover plate, a lower cover plate, and one or more sample slides, when there are multiple sample slides, which are stacked and installed between the upper cover plate and the lower cover plate; the upper cover plate and the lower cover plate have a sample interface and a sample exhaust port, which respectively communicate with the inner cavity inlet and the inner cavity outlet of the sample slide, and the inner cavity inlets and the inner cavity outlets between the multiple slides are interconnected; the sample slide has a through hole in the center for installing a working medium distributor, and the working medium distributor is arranged between the upper cover plate, the lower cover plate and the sample slide to form a multi-layer fixed gap; the upper cover plate has a cooling and rewarming working medium interface in the center. The application adopts a combined structure which can be conveniently disassembled, ensures that the cooling and rewarming working medium can perform high-speed cooling and rewarming, realizes quick change of the device flux, and through the superposition of the sample slide, milliliter-level sample cryopreservation can be realized, greatly improving the versatility and practicality of the low-temperature preservation device.

Description

Technical Field

[0001] This invention relates to the field of cryopreservation technology, specifically to a combined high-throughput, high-speed cooling and rewarming device for ultra-high-speed cooling and rewarming of large-throughput biological samples. Background Technology

[0002] Cryopreservation is currently the only effective long-term preservation method for many biological materials, and vitrification is an important method in biological cryopreservation. Rapid cooling to avoid the formation of ice crystals inside and outside cells is a necessary condition for vitrification. Furthermore, the rewarming rate required for the rewarming process is comparable to or higher than the critical cooling rate of the cooling process; insufficient rewarming may lead to cell recrystallization. Therefore, achieving rapid cooling and rewarming is of great significance for vitrification preservation. However, current vitrification preservation methods for biological samples suffer from one or more of the following problems: low cooling and rewarming rates, the need for high concentrations of cryoprotectants, low cryopreservation throughput, difficulty in increasing cryopreservation throughput to the milliliter level or the inability to change cryopreservation throughput, and specific samples requiring specific throughput consumables. These problems may lead to a narrow applicability of vitrification preservation methods, making them difficult to apply to the cryopreservation of various biological samples. Summary of the Invention

[0003] The purpose of this invention is to provide a high-throughput, high-speed de-freezing and rewarming device that allows for flexible adjustment of the loading capacity. This device provides a new de-freezing and rewarming device structure that can flexibly change the throughput of sample cryopreservation, while also providing a convenient and pollution-free method for loading biological samples.

[0004] To achieve the above objectives, the present invention adopts the following technical solution: a combined high-throughput, high-speed cooling and reheating device, comprising an upper cover plate, a lower cover plate, and one or more sample carriers; the sample carriers are flat and hollow, with an inner cavity for carrying samples; when multiple sample carriers are present, they are stacked between the upper and lower cover plates, and the inner cavity outlet of each sample carrier is sequentially connected to the inner cavity inlet of the sample carrier below it; the upper cover plate has a cooling and reheating working fluid interface and a sample interface at its center, and the lower cover plate has a sample exhaust port at its bottom; the sample interface is connected to the inner cavity inlet of the sample carrier below it, and the sample exhaust port is connected to the inner cavity outlet of the sample carrier above it; when the sample is injected through the sample interface, each sample carrier is sequentially filled. The sample carrier has a through hole in its center for mounting a working fluid distributor. The working fluid distributor has a normal flow guide ring and a tangential flow guide structure, with a through structure at its center. The normal flow guide ring is installed in the central through hole of the sample carrier, while the tangential flow guide structure is located between each sample carrier and the upper and lower cover plates, creating a gap of a specific height. The through structure in the center of each working fluid distributor is aligned and connected to the cooling / reheating working fluid interface of the upper cover plate. When the cooling / reheating working fluid is injected through the cooling / reheating working fluid interface, it will sequentially enter each working fluid distributor and be distributed to the upper and lower surfaces of the sample carrier via the tangential flow guide structure, simultaneously cooling or reheating the samples in all sample carriers.

[0005] The sample interface of the upper cover plate and the sample exhaust port of the lower cover plate are arranged symmetrically on the outer corners of the upper and lower cover plates respectively; the center of the upper cover plate is a through structure, which is connected to the cooling and reheating working medium interface, and an upper positioning groove for positioning the working medium distributor is provided below the center; the center of the lower cover plate is a closed structure, and a lower positioning groove for positioning the working medium distributor is provided above it.

[0006] The sample carrier includes a first heat exchange plate and a second heat exchange plate symmetrically arranged, as well as a sealed inner frame and a sealed outer frame in the middle. The hollow structure of the sample carrier is formed by the sealed inner frame and the sealed outer frame forming an annular flow channel, and the first heat exchange plate and the second heat exchange plate are tightly combined with it to form an annular chamber. The inlet of the inner chamber is located at the corner of the first heat exchange plate, and the outlet of the inner chamber is located at the corner of the second heat exchange plate, with the positions being centrally symmetrical with respect to the inlet of the inner chamber.

[0007] The first heat exchange plate, the second heat exchange plate, the middle sealing inner frame and the sealing outer frame are all made of high thermal conductivity materials, and the outer surfaces of the first heat exchange plate and the second heat exchange plate are all subjected to enhanced heat exchange treatment.

[0008] The tangential flow guide structure of the working fluid distributor is a radial flow channel structure with a positioning boss at the top; the normal flow guide ring of the working fluid distributor is a ring structure with a groove, the positioning boss is positioned and connected to the upper positioning groove of the upper cover plate or the groove of the normal flow guide ring of the upper working fluid distributor, and the groove of the normal flow guide ring is positioned and connected to the lower positioning groove of the lower cover plate or the positioning boss of the upper working fluid distributor.

[0009] The beneficial effects of this invention patent are:

[0010] 1) The sample slides of the present invention can be quickly combined and stacked to create different throughput inner cavity volumes to hold different throughput biological samples according to needs, which has higher flexibility and convenience compared with traditional biological sample cryopreservation methods.

[0011] 2) This invention enables the rapid loading of biological samples using syringes and other means. Combined sample slides can be gradually filled through interconnected flow channels without the need for individual filling, reducing the risk of contamination during operation and improving the convenience of loading and extraction. Attached Figure Description

[0012] Figure 1 This is a schematic diagram of the assembly structure of the present invention;

[0013] Figure 2 This is an exploded view of the present invention;

[0014] Figure 3 This is a schematic diagram of the structure of the upper cover plate of the present invention. Figure 1 ;

[0015] Figure 4 This is a schematic diagram of the structure of the upper cover plate of the present invention. Figure 2 ;

[0016] Figure 5 This is a schematic diagram of the structure of the lower cover plate of the present invention. Figure 1 ;

[0017] Figure 6 This is a schematic diagram of the structure of the lower cover plate of the present invention. Figure 2 ;

[0018] Figure 7 This is a schematic diagram of the structure of the sample carrier of the present invention;

[0019] Figure 8 yes Figure 7 An explosion diagram;

[0020] Figure 9 This is a schematic diagram of the working fluid distributor of the present invention. Figure 1 ;

[0021] Figure 10 This is a schematic diagram of the working fluid distributor of the present invention. Figure 2 ;

[0022] Figure 11 This is a schematic diagram of the multi-sample carrier assembly structure of the present invention;

[0023] The markings in the above figures are as follows: upper cover plate 1, cooling and reheating working fluid interface 11, sample interface 12, upper positioning groove 13, sample carrier plate 2, first heat exchange plate 21, second heat exchange plate 22, inner cavity inlet 23, inner cavity outlet 24, sealing inner frame 25, sealing outer frame 26, lower cover plate 3, sample exhaust port 31, lower positioning groove 32, working fluid distributor 4, tangential flow guide structure 41, positioning boss 42, normal flow guide ring 43. Detailed Implementation

[0024] The present invention will be further described below with reference to the accompanying drawings:

[0025] like Figure 1 , Figure 2 The illustrated combined high-throughput, high-speed cooling and rewarming device includes an upper cover plate 1, a lower cover plate 3, and a sample slide 2. The sample slide 2 has a through hole in its center and is fixedly held in the middle by the upper cover plate 1 and the lower cover plate 3. A working fluid distributor 4 is inserted into the through hole in the center of the sample slide 2 and is cross-assembled with the upper cover plate 1, the lower cover plate 3, and the sample slide 2. The working fluid distributor 4 has a normal flow guide ring 43 and a tangential flow guide structure 41, and its middle part is a through structure. The tangential flow guide structure 41 is used to form a specific assembly gap between the upper cover plate 1, the lower cover plate 3, and the sample slide 2. The upper cover plate 1 has a cooling and rewarming working fluid interface 11 in its center. The cooling and rewarming working fluid enters the working fluid distributor 4 through the cooling and rewarming working fluid interface 11, and enters the assembly gaps of each layer through the normal flow guide ring 43 and the tangential flow guide structure 41 to exchange heat with the sample slide 2 at high speed. A sample interface 12 is provided at the corner of the upper cover plate 1 for injecting biological samples. The lower cover plate 3 has a sample exhaust port 31 at its corner, which is used to expel air from the sample slide 2 when a sample is injected. The sample slide 2 has an inner cavity inlet 23 and an inner cavity outlet 24, which are aligned and connected to the sample interface 12 of the upper cover plate 1 and the sample exhaust port 31 of the lower cover plate 3, respectively.

[0026] Furthermore, such as Figure 3 , Figure 4 As shown, a cooling / reheating working medium interface 11 is provided at the center of the upper cover plate 1. The cooling / reheating working medium interface 11 is a through hole. An upper positioning groove 13 is provided below the upper cover plate 1. The position of the working medium distributor 4 is fixed by the upper positioning groove 13, and the external cooling / reheating working medium is introduced through the cooling / reheating working medium interface 11 and enters the working medium distributor 4 through the through hole. A sample interface 12 is provided at the corner of the upper cover plate 1. The sample interface 12 is sealed with an elastic material and can be inserted by a syringe needle to inject biological samples. The sample interface 12 corresponds to the inner cavity inlet 23 of the sample slide 2, through which the biological sample is injected into the sample slide 2.

[0027] Furthermore, such as Figure 5 , Figure 6 As shown, a lower positioning groove 32 is provided at the center of the upper part of the lower cover plate 3 to position the working fluid distributor 4. The lower positioning groove 32 is closed in the center, so that the cooling and reheating working fluid introduced into the working fluid distributor 4 can only flow out through the tangential flow guide structure 41 set by the working fluid distributor 4. Sample exhaust ports 31 are provided at the corners of the lower cover plate 3. The sample exhaust ports 31 are sealed with elastic material, and their upper parts correspond to the inner cavity outlet 24 of the sample slide 2. When injecting the sample, the needle is inserted, and the excess air in the sample slide 2 can be discharged through this outlet.

[0028] Furthermore, such as Figure 7 , Figure 8 The sample carrier 2 shown is a hollow structure, including a first heat exchange plate 21, a second heat exchange plate 22, and a sealed inner frame 25 and a sealed outer frame 26 in between. The sealed inner frame 25 and the sealed outer frame 26 form a flow channel. The first heat exchange plate 21 and the second heat exchange plate 22 close the upper and lower surfaces of the sealed inner frame 25 and the sealed outer frame 26 to form an internal cavity. The sample carrier 2 has a through hole in the center for inserting and assembling the working fluid distributor 4. The first heat exchange plate 21 is provided with an inner cavity inlet 23, which connects to the sample interface 1 of the upper cover plate 1. The second heat exchange plate 22 is aligned and connected, with an inner cavity outlet 24 that is aligned and connected to the sample exhaust port 31 of the lower cover plate 3. The inner cavity inlet 23 and the inner cavity outlet 24 are centrally symmetrically arranged. When injecting samples, the samples enter through the inner cavity inlet 23, gradually filling the hollow structure and expelling excess gas through the aligned inner cavity outlet 24. When setting up multi-stage sample carriers 2, the inner cavity outlet 24 of the previous stage sample carrier 2 is connected to the inner cavity inlet 23 of the next stage sample carrier 2, thus realizing the step-by-step filling of each stage of the sample carrier 2. The outer surfaces of the first heat exchange plate 21 and the second heat exchange plate 22 can be subjected to enhanced heat exchange treatment to improve the cooling and reheating effect.

[0029] Furthermore, such as Figure 9 , Figure 10 The working fluid distributor 4 shown has a normal flow guide ring 43 and a tangential flow guide structure 41. The tangential flow guide structure 41 has a radial flow channel structure and a positioning boss 42 on the top, which is positioned and connected to the upper positioning groove 13 of the upper cover plate 1. The normal flow guide ring 43 has a groove structure, which is positioned and connected to the lower positioning groove 32 of the lower cover plate 3. The working fluid distributors 4 are installed together and positioned by the positioning boss 42 and the groove structure of the normal flow guide ring 43. The stacked working fluid distributors 4 form a specific gap between the upper cover plate 1, the lower cover plate 3 and the sample carrier 2 through the tangential flow guide structure 41. The cooling and recooling working fluid introduced by the cooling and recooling working fluid interface 11 of the upper cover plate 1 is introduced through the normal flow guide ring 43 and flows out radially at high speed through the gap formed by the tangential flow guide structure 41 to exchange heat with the surface of each sample carrier 2 at high speed.

[0030] Furthermore, such as Figure 11As shown, when combining multiple sample carriers 2, only the inner cavity outlet 24 and inner cavity inlet 23 of the corresponding sample carrier 2 and the working fluid distributor 4 assembled in the center are required.

[0031] The working principle of this invention is as follows:

[0032] (1) Modular variable high-throughput structure. The overall throughput can be quickly changed through specially designed sample slides and cross-mounted working fluid dispensers, greatly increasing applicability and flexibility. Milliliter-level sample cryopreservation can be achieved by stacking sample slides.

[0033] (2) Pollution-free continuous filling. The sample slides are loaded into the shell through continuous flow channels, eliminating the need for individual filling combinations and reducing operational risks.

[0034] The cooling and reheating process of this invention can be controlled and operated based on a working fluid drive and control system. The specific structure of the working fluid drive and control system can be found in the prior art, and will not be described in detail here.

[0035] The cooling and reheating process of this invention is as follows:

[0036] First, prepare the cell solution and sterile syringe for the upcoming test, and use the sterile syringe to draw the required amount of cell solution for later use.

[0037] Second, insert the syringe filled with cell fluid into the sample port 12 of the upper cover plate 1, and insert the needle of a syringe or infusion tube into the sample vent 31 of the lower cover plate 3 to facilitate the expulsion of air during injection.

[0038] Third, gently push in the required amount of cell fluid. Stop when you see liquid appearing at the sample vent 31 of the lower cover plate 3. Then remove the needle from the sample vent 31 and pull out the syringe from the sample interface 12 of the upper cover plate 1.

[0039] Fourth, prepare the working fluid drive control system and liquid nitrogen required for the experiment, connect the working fluid interface 11 of the combined high-throughput high-speed cooling and reheating device to the end of the liquid nitrogen jet pipeline of the working fluid drive control system, and then place an insulation box at the bottom to collect the liquid nitrogen of the jet.

[0040] Fifth, turn on the pipeline of the working fluid drive and control system to complete the jet impact cooling process. Then, separate the liquid nitrogen tank line from the cooling and reheating working fluid interface 11 of the combined high-throughput high-speed cooling and reheating device, remove it from the pipeline and drop it into the liquid nitrogen in the bottom insulation box, thus completing the cooling experiment process.

[0041] Sixth, use tweezers or similar tools to adjust the position of the combined high-throughput high-speed cooling and reheating device in the insulation box so that it is aligned with the reheating pipeline of the working fluid drive control system and connected, and use the box body for temporary insulation.

[0042] Seventh, prepare the working fluid drive control system and the reheating working fluid required for the experiment, then turn on the pipeline of the working fluid drive and control system to complete the jet impact reheating process. Then separate the reheating pipeline from the cooling and reheating working fluid interface 11 of the combined high-throughput high-speed cooling and reheating device, and let it fall into the heat preservation box containing the reheating working fluid for heat preservation.

[0043] Eighth, take out the combined high-throughput high-speed cooling and reheating device from the insulated box, wipe off the surface moisture, prepare a sterile syringe and needle, insert the sterile syringe into the sample interface 12 on the upper cover plate 1, and insert the needle into the sample exhaust port 31 on the lower cover plate 3.

[0044] Ninth, slowly extract the sample from the combined high-throughput high-speed rewarming device by pulling the sterile syringe until all samples are extracted. Then, pull out the sterile syringe, remove the needle from the sample exhaust port 31, transfer the cell fluid in the syringe, and complete the cell rewarming experiment.

[0045] Example 1:

[0046] In this embodiment, the upper cover plate 1 and the lower cover plate 3 are made of PMMA, with an overall size of 45*45*5mm. The inner diameter of the cooling and reheating working fluid interface 11 of the upper cover plate 1 is 5mm. The upper positioning groove has a diameter of 8mm and a depth of 0.3mm. The lower positioning groove of the lower cover plate has a diameter of 9.1mm and a depth of 0.6mm. The first and second heat exchange plates 22 of the sample carrier 2 are made of aluminum nitride ceramic sheets with a size of 35*35*0.2mm. The surface heat exchange enhancement treatment is a fork-shaped biomimetic microchannel with a channel depth of 0.1mm. The sealing inner frame 25 and the sealing outer frame 26 in the middle of the sample carrier 2 are also made of aluminum nitride ceramic sheets. The hollow structure has an outer diameter of 29 mm, an inner diameter of 15 mm, and a thickness of 0.3 mm. The overall size of the sample slide is 35*35*0.6 mm, and it can hold approximately 0.15 ml of cell fluid. The diameter of the through hole in the middle of the sample slide 2 is 9.1 mm. The working fluid dispenser 4 has two layers. The tangential flow guide structure 41 has a flow channel height of 0.8 mm and a maximum diameter of 15 mm. The positioning boss 42 has a height of 0.3 mm and a diameter of 7.8 mm. The normal flow guide ring 43 has an outer diameter of 9 mm, a height of 0.6 mm, a groove diameter of 8 mm, a groove depth of 0.3 mm, and a central through hole diameter of 5 mm.

[0047] As described above, this embodiment uses one sample slide 2 with a layer height of 0.8 mm for the working fluid flow on both sides, capable of holding 0.15 ml of cell fluid for cooling and rewarming tests. Statistical analysis of the test results shows that the average cooling rate in this embodiment reaches 190℃ / s, and the rewarming rate reaches 960℃ / s.

[0048] Example 2:

[0049] like Figure 11 As shown, to increase the throughput of biological samples used in each cooling and rewarming cycle, the sample slides 2 of this invention are assembled into three pieces; correspondingly, the number of working fluid distributors 4 is increased to four, with one working fluid distributor 4 arranged between each pair of the upper cover plate 1, the lower cover plate 3, and the sample slides 2, and the sample interfaces 12, inner cavity inlets 23, inner cavity outlets 24, and sample exhaust ports 31 of each stage are installed accordingly. After increasing the throughput, the cooling and rewarming working fluid will be introduced into the surface of all sample slides 2 by the working fluid distributors 4 to achieve a high-speed cooling and rewarming effect.

[0050] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. A combined high-throughput, high-speed cooling and reheating device, characterized in that: Includes an upper cover plate (1), a lower cover plate (3), and multiple sample carriers (2); the sample carriers (2) are flat and hollow, with an inner cavity for carrying samples; multiple sample carriers (2) are stacked between the upper cover plate (1) and the lower cover plate (3), and the inner cavity outlet (24) of each sample carrier (2) is sequentially connected to the sample inner cavity inlet (23) below it; the upper cover plate (1) has a sample interface (12) above it, and the lower cover plate (3) has a sample exhaust port (31) below it. The sample interface (12) is connected to the inner cavity inlet (23) of the sample carrier (2) below it, and the sample exhaust port (31) is connected to the inner cavity outlet (24) of the sample carrier (2) above it, forming a continuous sample path. When the sample is injected through the sample interface (12), each sample carrier (2) is sequentially filled; the sample carrier (2) has a through hole in the center for installing a working fluid dispenser (4). The working fluid distributor (4) has a normal flow guide ring (43) and a tangential flow guide structure (41). The normal flow guide ring (43) is installed in the central through hole of the sample slide, while the tangential flow guide structure (41) is located between each sample slide (2) and the upper cover plate (1) and the lower cover plate (3), separating each sample slide (2) from the upper cover plate (1) and the lower cover plate (3) to form a gap of a specific height. The upper cover plate (1) has a cooling and reheating working fluid interface (11) in the center, which is aligned and connected with the normal flow guide ring (43) of each working fluid distributor (4). When the cooling and reheating working fluid is injected through the cooling and reheating working fluid interface (11), it will enter each working fluid distributor (4) in sequence and be distributed to the upper and lower surfaces of the sample slide (2) through the tangential flow guide structure (41). The samples in all sample slides (2) are simultaneously cooled or reheated by high-speed shear flow on the surface of the sample slide (2).

2. The combined high-throughput high-speed cooling and reheating device according to claim 1, characterized in that: The sample interface (12) of the upper cover plate (1) and the sample exhaust port (31) of the lower cover plate (3) are arranged symmetrically on the outer corners of the upper cover plate (1) and the lower cover plate (3), respectively. The center of the upper cover plate (1) is a through structure, which is connected to the cooling and reheating working medium interface (11). An upper positioning groove (13) for positioning the working medium distributor (4) is provided below the center. The center of the lower cover plate (3) is a closed structure, and a lower positioning groove (32) for positioning the working medium distributor (4) is provided above it.

3. The combined high-throughput high-speed cooling and reheating device according to claim 1, characterized in that: The sample carrier (2) includes a first heat exchange plate (21) and a second heat exchange plate (22) arranged symmetrically, as well as a sealed inner frame (25) and a sealed outer frame (26) in the middle. The hollow structure of the sample carrier forms an annular flow channel by the sealed inner frame (25) and the sealed outer frame (26), and the first heat exchange plate (21) and the second heat exchange plate (22) are tightly connected with the upper and lower surfaces of the annular flow channel formed by the sealed inner frame (25) and the sealed outer frame (26) to form an annular chamber. The inner cavity inlet (23) is located at the corner of the first heat exchange plate (21), and the inner cavity outlet (24) is located at the corner of the second heat exchange plate (22), with the positions being centrally symmetrical with the inner cavity inlet (23).

4. The combined high-throughput high-speed cooling and reheating device according to claim 3, characterized in that: The first heat exchange plate (21), the second heat exchange plate (22), the middle sealing inner frame (25) and the sealing outer frame (26) are all made of high thermal conductivity materials, and the outer surfaces of the first heat exchange plate (21) and the second heat exchange plate (22) are all subjected to enhanced heat exchange treatment.

5. The combined high-throughput high-speed cooling and reheating device according to claim 1, characterized in that: The tangential flow guide structure (41) of the working fluid distributor (4) is a radial flow channel structure with a positioning boss (42) on the top; the normal flow guide ring (43) of the working fluid distributor (4) is a ring structure with a groove. The positioning boss (42) is positioned and connected to the upper positioning groove (13) of the upper cover plate (1) or the groove of the normal flow guide ring (43) of the upper working fluid distributor (4). The groove of the normal flow guide ring (43) is positioned and connected to the lower positioning groove (32) of the lower cover plate (3) or the positioning boss (42) of the upper working fluid distributor (4).

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