Rapid freezing quenching device and mixed injection mechanism and rapid freezing quenching method thereof

Abstract

The invention relates to a rapid freezing quenching device, a mixed injection mechanism thereof and a rapid freezing quenching method. The mixing and spraying mechanism comprises a mixing component and a nozzle. The mixing component is provided with a first convection flow channel, a second convection flow channel and a converging flow channel, one end of the first convection flow channel is connected with one end of the second convection flow channel and converged at one end of the converging flow channel, the first convection flow channel is used for being communicated with the first injection component, and the second convection flow channel is used for being communicated with the second injection component; and the nozzle is communicated with the other end of the converging runner. In the mixing part, all reaction liquids can be rapidly and fully mixed after being contacted, and the cavity structure of the mixing part has a relatively small dead volume, so that the dead time in the sample preparation process is effectively reduced, rapid conveying is realized while the samples are mixed, the mixture is sprayed out at a high speed through the nozzle, the flight time is reduced, and the sample preparation efficiency is improved. The time resolution is improved, and the intermediate in the rapid reaction process can be better captured.

Description

Technical Field

[0001] The present invention relates to the technical field of biochemical experimental equipment, and particularly relates to a rapid freeze quench device, its mixing and spraying mechanism, and a rapid freeze quench method. Background Art

[0002] A rapid freeze quench (RFQ) device is a sample preparation device for studying biochemical reaction kinetics, and particularly plays an important role in the research on the structure and properties of intermediates in rapid reaction processes. The rapid freeze quench device rapidly stops the ongoing chemical reaction or biological process by reducing the temperature of the sample to an extremely low level in an extremely short time.

[0003] The core requirements of the rapid freeze quench device include: 1. Each sample can be rapidly and fully mixed; 2. After the samples are mixed and ejected, they can be rapidly frozen and formed by a refrigerant; 3. The frozen and formed samples can be efficiently collected for subsequent testing; 4. The mixing and freezing time of the samples can be precisely controlled (accurate to milliseconds or even shorter), that is, it has a higher time resolution.

[0004] For a rapid freeze quench device, a higher time resolution means that the reaction process can be observed more carefully. The time resolution of traditional rapid freeze quench devices (such as the SFM-3000 of Bio-Logic) is generally at the millisecond level, which limits the traditional rapid freeze quench devices in studying some key scientific problems in disciplines such as protein folding kinetics, enzyme kinetics, and rapid chemical reaction kinetics. Summary of the Invention

[0005] Based on this, it is necessary to provide a rapid freeze quench device, its mixing and spraying mechanism, and a rapid freeze quench method to improve the time resolution of the device.

[0006] The first aspect of the present invention is to provide a mixing and spraying mechanism for a rapid freeze quench device, and the solution is as follows:

[0007] A mixing and spraying mechanism for a rapid freeze quench device includes a mixing component and a nozzle;

[0008] The mixing component is provided with a first convection channel, a second convection channel and a confluence channel. One end of the first convection channel is connected to one end of the second convection channel and converges at one end of the confluence channel. The other end of the first convection channel is used to communicate with a first injection component, the other end of the second convection channel is used to communicate with a second injection component, and the nozzle is communicated with the other end of the confluence channel.

[0009] In one embodiment, both the first convection channel and the second convection channel are linear channels, and the included angle is 150° - 180°.

[0010] In one embodiment, the confluence channel is a linear channel, and the included angles between the confluence channel and the first convection channel and the second convection channel are 90° - 105°.

[0011] In one embodiment, the length of the first convection channel is 0.5 cm - 1 cm, and the inner diameter is 0.5 mm - 1 mm.

[0012] In one embodiment, the length of the second convection channel is 0.5 cm - 1 cm, and the inner diameter is 0.5 mm - 1 mm.

[0013] In one embodiment, the length of the confluence channel is 0.5 cm - 1 cm, and the inner diameter is 0.5 mm - 1 mm.

[0014] In one embodiment, the caliber of the nozzle is 0.1 mm - 0.2 mm.

[0015] In one embodiment, the mixing and spraying mechanism includes a plurality of nozzles with different calibers, and each nozzle is detachably connected to the mixing component.

[0016] In one embodiment, the mixing and spraying mechanism includes nozzles with calibers of 0.1 mm, 0.15 mm and 0.2 mm respectively.

[0017] In one embodiment, the mixing and spraying mechanism further includes a first injection component and a second injection component. The first injection component is communicated with the first convection channel through a first sample delivery pipeline, and the second injection component is communicated with the second convection channel through a second sample delivery pipeline.

[0018] In one embodiment, the specification of the first injection component is 500 μL - 2 mL.

[0019] In one embodiment, the specification of the second injection component is 500 μL - 2 mL.

[0020] In one embodiment, the inner diameter of the first sample delivery pipe is 0.13 mm to 0.5 mm.

[0021] In one embodiment, the inner diameter of the second sample delivery pipe is 0.13 mm to 0.5 mm.

[0022] In one embodiment, the mixing and injection mechanism further includes a pushing member for pushing the first injection member and the second injection member to perform synchronous injection sampling.

[0023] In one embodiment, the pushing speed of the pushing member can reach 100 mm / s, and the displacement accuracy is 0.01 mm to 0.02 mm.

[0024] In one embodiment, the pushing member is an electric push rod.

[0025] The second aspect of the present invention is to provide a rapid freezing and quenching device, and the solution is as follows:

[0026] A rapid freezing and quenching device includes the mixing and injection mechanism described in any one of the above embodiments and a freezing mechanism for receiving the mixture ejected from the nozzle and freezing the mixture.

[0027] In one embodiment, the freezing mechanism includes a temperature reduction member, a receiving and freezing member, and a rotation driving member. The receiving and freezing member is located below the nozzle for receiving the mixture ejected from the nozzle. The temperature reduction member is used to refrigerate the receiving and freezing member, and the rotation driving member is connected to the receiving and freezing member to drive the receiving and freezing member to rotate.

[0028] In one embodiment, an annular groove is provided at the upper end of the receiving and freezing member around the rotation axis.

[0029] In one embodiment, the temperature reduction member includes a cooling medium container with an opening at the upper end. The receiving and freezing member extends into the cooling medium container through the opening, and the size of the receiving and freezing member gradually decreases from the upper end to the lower end.

[0030] In one embodiment, the freezing mechanism further includes a cover box, and the mixing member, the nozzle, and the receiving and freezing member are all housed in the cover box.

[0031] The third aspect of the present invention is to provide a rapid freezing and quenching method, and the solution is as follows:

[0032] A rapid freezing and quenching method uses the rapid freezing and quenching device described in any one of the above embodiments, and the rapid freezing and quenching method includes the following steps:

[0033] Samples are respectively injected into the first convective flow channel and the second convective flow channel of the mixing component. The samples are mixed and ejected through the confluent flow channel and the nozzle onto the freezing mechanism for freeze forming.

[0034] Compared with the traditional technology, the above-mentioned rapid freezing and quenching device, its mixing and injection mechanism, and the rapid freezing and quenching method have the following beneficial effects:

[0035] The above-mentioned mixing and injection mechanism is provided with a mixing component to mix different samples. One end of the first convective flow channel and one end of the second convective flow channel in the mixing component are connected and converge at one end of the confluent flow channel, so that the reaction solutions of all paths can be mixed quickly and fully after contact. The cavity structure of the mixing component has a small dead volume, thus effectively reducing the dead time in the sample preparation process, and realizing rapid transportation while mixing the samples. The above-mentioned mixing and injection mechanism is also provided with a nozzle communicated with the confluent flow channel, and the mixture is ejected at a high speed through the nozzle, reducing the flight time, improving the time resolution of the device, and being able to better capture the intermediate in the rapid reaction process.

[0036] The above-mentioned rapid freezing and quenching device includes the mixing and injection mechanism described in any one of the above embodiments, and thus has corresponding technical features and can obtain corresponding beneficial effects. The above-mentioned rapid freezing and quenching method uses the rapid freezing and quenching device described in any one of the above embodiments for freezing and quenching, and can obtain corresponding beneficial effects. Description of the Drawings

[0037] Figure 1 It is a schematic structural diagram of a mixing and injection mechanism of an embodiment;

[0038] Figure 2 is Figure 1 a schematic structural diagram of the mixing component and the nozzle in the mixing and injection mechanism shown;

[0039] Figure 3 is an embodiment including Figure 1 a schematic structural diagram of a rapid freezing and quenching device with the mixing and injection mechanism shown;

[0040] Figure 4 is Figure 3 a schematic structural diagram of the receiving freezing component, the rotation driving component and the connecting rod in the rapid freezing and quenching device shown.

[0041] Description of the Reference Numerals:

[0042] 100, mixing injection mechanism; 110, first injection component; 120, second injection component; 130, mixing component; 131, first convection channel; 132, second convection channel; 133, converging channel; 134, first connecting hole; 135, second connecting hole; 136, third connecting hole; 140, nozzle; 150, first sample delivery pipeline; 160, second sample delivery pipeline; 170, pushing component; 180, position adjustment component; 181, vertical adjustment member; 182, horizontal adjustment member; 183, optical cross fixing clamp; 10, rapid freezing quenching device; 200, freezing mechanism; 210, cooling component; 211, cooling medium container; 212, cooling medium delivery pipeline; 220, freezing receiving component; 221, annular groove; 230, rotating driving component; 240, connecting rod; 250, cover box. DETAILED DESCRIPTION

[0043] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the specific embodiments of the present invention are described in detail below in conjunction with the accompanying drawings. In the following description, many specific details are set forth to facilitate a full understanding of the present invention. However, the present invention can be implemented in many other ways different from those described herein, and those skilled in the art can make similar improvements without violating the connotation of the present invention, so the present invention is not limited by the specific embodiments disclosed below.

[0044] In the description of the present invention, it should be understood that the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside", "clockwise", "counterclockwise", "axial", "radial", "circumferential" and the like indicate orientations or positional relationships based on the orientations or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present invention and simplifying the description, and do not indicate or imply that the referred device or element must have a specific orientation, be constructed and operated in a specific orientation, and therefore should not be understood as limiting the present invention.

[0045] In addition, the terms "first" and "second" are used for descriptive purposes only and should not be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features. Therefore, the features defined as "first" and "second" may explicitly or implicitly include at least one of the features. In the description of the present invention, the meaning of "plurality" is at least two, such as two, three, etc., unless otherwise clearly and specifically defined.

[0046] In the present invention, unless otherwise clearly defined or limited, terms such as "installed", "connected", "joined", "fixed", etc. shall be construed broadly. For example, it may be a fixed connection, a detachable connection, or an integral one; it may be a mechanical connection or an electrical connection; it may be directly connected or indirectly connected through an intermediate medium, and it may be the internal communication between two elements or the interaction relationship between two elements, unless otherwise clearly defined. For those of ordinary skill in the art, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0047] In the present invention, unless otherwise clearly defined or limited, the first feature being "on" or "under" the second feature may be that the first and second features are in direct contact, or the first and second features are indirectly in contact through an intermediate medium. Moreover, the first feature being "above", "over" and "on top of" the second feature may be that the first feature is directly above or obliquely above the second feature, or merely indicates that the first feature has a higher horizontal height than the second feature. The first feature being "under", "beneath" and "underneath" the second feature may be that the first feature is directly below or obliquely below the second feature, or merely indicates that the first feature has a lower horizontal height than the second feature.

[0048] It should be noted that when an element is referred to as "fixed to" or "disposed on" another element, it may be directly on the other element or there may also be an intermediate element. When an element is considered to be "connected" to another element, it may be directly connected to the other element or there may be an intermediate element at the same time. The terms "vertical", "horizontal", "up", "down", "left", "right" and similar expressions used herein are only for the purpose of illustration and do not represent the only implementation.

[0049] In the description of the present invention, the meaning of "several" is more than one, the meaning of "multiple" is more than two, and understandings such as "greater than", "less than", "exceeding", etc. do not include the present number, and understandings such as "above", "below", "within", etc. include the present number. If there is a description of "first" and "second", it is only for the purpose of distinguishing technical features and cannot be understood as indicating or implying relative importance or implicitly indicating the quantity of the indicated technical features or implicitly indicating the sequence relationship of the indicated technical features.

[0050] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the technical field to which the present invention belongs. The terms used herein in the description of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention. The term "and / or" used herein includes any and all combinations of one or more of the related listed items.

[0051] The present invention provides a hybrid injection mechanism and a rapid freezing quenching device including the hybrid injection mechanism. The hybrid injection mechanism is used to quickly mix different reactants and eject them to a freezing mechanism for instant freezing and molding. The above-mentioned rapid freezing quenching device can be used to prepare samples at a specific time point during a rapid reaction process, for example, it can be used to capture and characterize transient, short-lived paramagnetic intermediates in biochemical reactions.

[0052] Combined Figure 1 with Figure 2 As shown, the hybrid injection mechanism 100 of an embodiment of the present invention includes a mixing component 130 and a nozzle 140.

[0053] As Figure 2 shown, the mixing component 130 is provided with a first convection channel 131, a second convection channel 132, and a converging channel 133. One end of the first convection channel 131 and one end of the second convection channel 132 are connected and converge at one end of the converging channel 133. The other end of the first convection channel 131 is used to communicate with a first injection component, and the other end of the second convection channel 132 is used to communicate with a second injection component. The nozzle 140 is communicated with the other end of the converging channel 133.

[0054] The above-mentioned hybrid injection mechanism 100 sets the mixing component 130 to mix different samples. One end of the first convection channel 131 and one end of the second convection channel 132 in the mixing component 130 are connected and converge at one end of the converging channel 133, so that the reaction liquids of each path can be quickly and fully mixed after contact. The cavity structure of the mixing component 130 has a small dead volume, thus effectively reducing the dead time in the sample preparation process and achieving rapid transportation while the samples are being mixed. The above-mentioned hybrid injection mechanism 100 is also provided with a nozzle 140 communicated with the converging channel 133, and the mixture is ejected at a high speed through the nozzle 140, reducing the flight time, improving the time resolution of the device, and being able to better capture the intermediates in the rapid reaction process.

[0055] The above-mentioned hybrid injection mechanism 100 also simplifies the steps of sample preparation, is beneficial to reducing the complexity and cost of experimental operations, improving the reliability and repeatability of experimental results, and providing strong technical support for research in related fields.

[0056] In some examples, the hybrid injection mechanism 100 further includes a first injection component 110 and a second injection component 120. The first injection component 110 and the second injection component 120 are used to inject different samples into the mixing component 130 respectively.

[0057] The materials of the first injection component 110 and the second injection component 120 can be, but are not limited to, glass. The first injection component 110 and the second injection component 120 can be installed on a bracket. For example, the first injection component 110 and the second injection component 120 are installed on an aluminum alloy bracket through detachable fasteners. The contact positions between the above injection components and the bracket can be wrapped with tape to prevent the injection components from being crushed due to stress during installation.

[0058] The first injection component 110 adopts a precision microinjector, which can accurately control the injection volume. For example, the specification of the first injection component 110 is, for example, 500 μL to 2 mL.

[0059] The second injection component 120 adopts a precision microinjector, which can accurately control the injection volume. For example, the specification of the second injection component 120 is, for example, 500 μL to 2 mL.

[0060] As Figure 1 shown, the first injection component 110 is communicated with the first convection channel 131 through the first sample delivery pipe 150. The second injection component 120 is communicated with the second convection channel 132 through the second sample delivery pipe 160.

[0061] In some examples, the lengths of the first sample delivery pipe 150 and the second sample delivery pipe 160 are 0.2 m to 0.5 m. In some examples, the inner diameters of the first sample delivery pipe 150 and the second sample delivery pipe 160 are 0.13 mm to 0.5 mm.

[0062] The materials of the first sample delivery pipe 150 and the second sample delivery pipe 160 can be, but are not limited to, polymers, specifically, for example, PEEK (polyetheretherketone). PEEK is relatively inert to biochemical reactions and will not react further with the components in the reaction system to interfere with the experimental results.

[0063] The needle parts of the above injection components and the corresponding sample delivery pipes can be connected through a first connecting component (not shown in the figure). The first connecting component includes a Luer connector, a gland, and a compression ring arranged between the Luer connector and the gland. The Luer connector is, for example, an M6 internal thread Luer connector, and the gland is, for example, an M6 threaded gland. The above first connecting component can improve the safety and accuracy of liquid delivery.

[0064] As Figure 1 shown, in some examples, the mixing and injection mechanism 100 further includes a pushing component 170, and the pushing component 170 is used to push the first injection component 110 and the second injection component 120 to perform synchronous injection sampling.

[0065] The pushing component 170 is installed on the bracket to prevent displacement during operation. For example, the pushing component 170 is installed on an aluminum alloy bracket through detachable fasteners.

[0066] The pushing component 170 preferably has high precision. For example, the pushing speed of the pushing component 170 can reach 100 mm / s, and the displacement precision of the pushing component 170 is 0.01 mm to 0.02 mm. The pushing component 170 can be, but is not limited to, an electric push rod. The electric push rod is preferably a high-precision electric push rod. For example, the fastest speed of the electric push rod can reach 100 mm / s. The displacement precision of the electric push rod is 0.01 mm to 0.02 mm, so that the pushing speeds of the first injection component 110 and the second injection component 120 can be accurately controlled, the accuracy of the injection volume of the above injection components can be improved, and the synchronous injection and sampling of the above injection components can also be ensured.

[0067] The power supply of the electric push rod uses a direct current (DC) power supply, and the output voltage is, for example, 24V ± 10%, and the output current is, for example, 5A.

[0068] The material of the mixing component 130 can be, but is not limited to, metal, specifically, for example, stainless steel. Stainless steel is relatively inert for biochemical reactions and will not react further with the components in the reaction system to interfere with the experimental results. The shape of the mixing component 130 can be, but is not limited to, a cuboid, for example, a cuboid with dimensions of 5 cm × 3 cm × 1 cm.

[0069] As Figure 2 As shown in (a) below, in some examples, the mixing component 130 is provided with a first connection hole 134 and a second connection hole 135. The first connection hole 134 communicates with the first convection channel 131 and is used to connect the first sample delivery pipe 150. The second connection hole 135 communicates with the second convection channel 132 and is used to connect the second sample delivery pipe 160.

[0070] In some examples, the first connection hole 134 and the second connection hole 135 have internal threads, such as M6 internal threads. The above connection holes and the corresponding sample delivery pipes can be connected through a second connecting component (not shown in the figure). The second connecting component includes a Luer connector, a gland, and a compression ring disposed between the Luer connector and the gland. The Luer connector is, for example, an M6 internal thread Luer connector, and the gland is, for example, an M6 threaded gland.

[0071] As Figure 2 As shown in (b) below, in some examples, the nozzle 140 is detachably connected to the mixing component 130, and the nozzle 140 can be replaced. In some examples, the nozzle 140 is threadedly connected to the mixing component 130. More specifically, the mixing component 130 is provided with a third connection hole 136, and the third connection hole 136 has an internal thread, such as M6 internal thread. The third connection hole 136 is in threaded engagement with the nozzle 140.

[0072] In some examples, both the first counterflow channel 131 and the second counterflow channel 132 are straight channels. In some examples, the included angle between the first counterflow channel 131 and the second counterflow channel 132 is 150° to 180°. Further, the included angle between the first counterflow channel 131 and the second counterflow channel 132 is 170° to 180°.

[0073] In some examples, the confluence channel 133 is a straight channel. The included angle between the confluence channel 133 and the first counterflow channel 131 and the second counterflow channel 132 is 90° to 105°. Further, the included angle between the confluence channel 133 and the first counterflow channel 131 and the second counterflow channel 132 is 90° to 95°.

[0074] In some examples, the length of the first counterflow channel 131 is 0.5 cm to 1 cm, and the inner diameter is 0.5 mm to 1 mm.

[0075] In some examples, the length of the second counterflow channel 132 is 0.5 cm to 1 cm, and the inner diameter is 0.5 mm to 1 mm.

[0076] In some examples, the length of the confluence channel 133 is 0.5 cm to 1 cm, and the inner diameter is 0.5 mm to 1 mm.

[0077] As Figure 1 shown, in some examples, the mixing injection mechanism 100 further includes a position adjustment component 180. The position adjustment component 180 is connected to the mixing component 130 to adjust the positions of the mixing component 130 and the nozzle 140, thereby adjusting the relative position between the nozzle 140 and the freezing mechanism. In some examples, the position adjustment component 180 can adjust the heights of the mixing component 130 and the nozzle 140, thereby adjusting the height difference between the nozzle 140 and the freezing mechanism, and further controlling the flight time of the mixture ejected from the nozzle 140. In some examples, the position adjustment component 180 can adjust the positions of the mixing component 130 and the nozzle 140 in the horizontal direction, thereby adjusting the relative position between the nozzle 140 and the freezing mechanism in the horizontal direction, and further controlling the landing point of the mixture on the freezing mechanism.

[0078] In some examples, the position adjustment component 180 includes a vertical adjustment member 181 and a horizontal adjustment member 182. The vertical adjustment member 181 can be welded to the aluminum alloy base. The vertical adjustment member 181 is, for example, a manual displacement stage. The horizontal adjustment member 182 is, for example, a metal rod with a diameter of, for example, 16 mm. The mixing component 130 is connected to the horizontal adjustment member 182. The horizontal adjustment member 182 is connected to the vertical adjustment member 181, for example, through an optical cross fixing clip 183 connected to the vertical adjustment member 181 to improve the accuracy of position adjustment.

[0079] The nozzle 140 has an extremely small caliber, which is smaller than that of the converging flow channel 133. For example, the caliber of the nozzle 140 is 0.1 mm to 0.2 mm. The extremely small caliber enables the reaction liquid after mixing to be ejected with a very high initial velocity and a short flight time. The mixture can be rapidly frozen within a time scale of sub-milliseconds, significantly improving the time resolution of the device, so as to better capture the intermediates in the fast reaction process.

[0080] In some examples, the mixing and ejection mechanism 100 includes a plurality of nozzles 140 with different calibers. Each nozzle 140 is detachably connected to the mixing component 130, and different-caliber nozzles 140 can be replaced according to needs. For example, the mixing and ejection mechanism 100 includes three replaceable nozzles 140 with calibers of 0.1 mm, 0.15 mm, and 0.2 mm respectively.

[0081] The above-mentioned mixing and ejection mechanism 100 sets a mixing component 130 to mix different samples. One end of the first countercurrent flow channel 131 and one end of the second countercurrent flow channel 132 in the mixing component 130 are connected and converge at one end of the converging flow channel 133, so that the reaction liquids of all paths can be quickly and fully mixed after contact. The cavity structure of the mixing component 130 has a small dead volume, thus effectively reducing the dead time in the sample preparation process and realizing fast transportation while mixing the samples. The above-mentioned mixing and ejection mechanism 100 is also provided with a nozzle 140 communicated with the converging flow channel 133. The mixture is ejected at a high speed through the nozzle 140, reducing the flight time, improving the time resolution of the device, and being able to better capture the intermediates in the fast reaction process.

[0082] As Figure 3 shown, the rapid freezing and quenching device 10 of an embodiment of the present invention includes the mixing and ejection mechanism 100 and the freezing mechanism 200 of any of the above examples. The freezing mechanism 200 is used to receive the mixture ejected from the nozzle 140 and freeze the mixture.

[0083] As Figure 3 shown, in some examples, the freezing mechanism 200 includes a cooling component 210 and a receiving and freezing component 220. Among them, the receiving and freezing component 220 is located below the nozzle 140 to receive the mixture ejected from the nozzle 140. The cooling component 210 is used to refrigerate the receiving and freezing component 220. When the mixture ejected from the nozzle 140 falls onto the receiving and freezing component 220, it is quickly frozen to quench the reaction.

[0084] The material for the receiving and freezing component 220 can be, but is not limited to, metal, specifically aluminum for example. By using the above-mentioned receiving and freezing component 220 for receiving, it can ensure that the flight distance of the mixture after ejection is fixed, avoiding the flight time difference caused by the liquid level change after the liquid refrigerant contacts the reaction liquid in the traditional method, and significantly improving the repeatability of the experiment.

[0085] In some examples, the freezing mechanism 200 further includes a rotation driving component 230. The rotation driving component 230 is connected to the receiving and freezing component 220 for driving the receiving and freezing component 220 to rotate. For example, the rotation driving component 230 is located above the receiving and freezing component 220 and is connected to the receiving and freezing component 220 through a connecting rod 240. During the working process, the rotation driving component 230 drives the receiving and freezing component 220 to rotate, preventing the mixture ejected from the nozzle 140 from accumulating at the same position on the receiving and freezing component 220 and affecting the freezing effect.

[0086] The rotation driving component 230 can be, but is not limited to, a motor. In some examples, the rotation driving component 230 is a constant-speed rotation motor.

[0087] As Figure 4 shown, in some examples, an annular groove 221 is provided at the upper end of the receiving and freezing component 220, which is arranged around the rotation axis for receiving the mixture ejected from the nozzle 140. The depth of the annular groove 221 is, for example, 1 mm to 2 mm. The number of the annular grooves 221 is not limited to only one, and there can also be multiple with different diameters.

[0088] In some examples, the temperature-lowering component 210 includes a cooling medium container 211 for containing a cooling medium such as liquid nitrogen. An opening is provided at the upper end of the cooling medium container 211, and the receiving and freezing component 220 extends into the cooling medium container 211 through the above-mentioned opening.

[0089] The cooling medium container 211 can be, but is not limited to, a Dewar flask. The cooling medium container 211 can be welded to a base made of aluminum alloy.

[0090] Furthermore, the temperature-lowering component 210 further includes a cooling medium delivery pipe 212. The cooling medium delivery pipe 212 is communicated with the cooling medium container 211, for example, connected to the side wall of the cooling medium container 211. During the working process, the cooling medium is continuously injected into the cooling medium container 211 through the cooling medium delivery pipe 212 to maintain the low-temperature state of the receiving and freezing component 220.

[0091] The material of the cooling medium delivery pipe 212 can be, but is not limited to, rubber, specifically latex for example.

[0092] In some examples, the size of the receiving freezing component 220 gradually decreases from the upper end to the lower end. The upper end of the receiving freezing component 220 is, for example, a circular tabletop with a diameter of, for example, 10 cm. In some examples, the receiving freezing component 220 is conical. The above structural design of the receiving freezing component 220 can improve the heat conduction cooling effect.

[0093] In some examples, the freezing mechanism 200 further includes a hood 250. The mixing component 130, the nozzle 140, and the receiving freezing component 220 are all housed in the hood 250. During operation, by continuously injecting high-purity nitrogen into the hood 250, the humidity inside the hood 250 can be reduced, the condensed water on the surface of the receiving freezing component 220 can be reduced, the frosting degree can be reduced, and the sample recovery rate can be improved.

[0094] The material of the hood 250 can be, but is not limited to, polymers, such as acrylic. The hood 250 is, for example, a bottomless cuboid with dimensions of, for example, 30 cm × 40 cm × 50 cm. A through hole, such as a circular hole with a diameter of 20 cm, can be provided on the hood 250 for the pipeline to pass through and for easy operation by personnel.

[0095] The rapid freezing quenching device 10 of the above embodiment can monitor the reaction time interval of 10 ms to 20 s by adjusting parameters such as the pushing speed of the pushing component 170, the caliber of the nozzle 140, the flight distance, and the injection component specifications.

[0096] It can be understood that the above components such as the pushing component 170 can be controlled by a control mechanism (not shown in the figure). The control mechanism is, for example, connected to the pushing component 170 through a network cable and communicates via the TCP protocol.

[0097] The parameters of each component at a specific reaction time can be calculated by the control mechanism, such as parameters such as the pushing speed of the pushing component 170, the caliber of the nozzle 140, and the flight distance (the distance between the nozzle 140 and the receiving freezing component 220).

[0098] The selection schemes of the speed of the pushing component 170, the caliber of the nozzle 140, and the flight distance are obtained by software calculation. The software is compiled by python and realizes interactive operations through a graphical user interface (GUI), and can accurately calculate parameters such as the linear speed of the electric push rod, the caliber specification of the nozzle 140, and the flight distance that need to be configured based on the reaction time input by the user.

[0099] The functions of the software include, for example, automatically selecting the optimal parameter combination, such as the pushing speed of the pushing component 170 and the caliber specification of the nozzle 140, according to the set time range (10-2000 milliseconds) and experimental conditions, and outputting the corresponding calculation results. The slider and button provide a flexible time control method, allowing users to accurately adjust the target time in units of 100 microseconds. The result interface is optimized for display, and different colors are used to distinguish parameter labels and calculation results to improve the readability of the interface. Parameterized dynamic calculations are realized, including core indicators such as flow rate, outlet velocity, and flight time, to meet the needs of various experimental configurations.

[0100] The software process includes the following steps:

[0101] Constant definition steps: define constants such as nozzle 140 diameter, pipe diameter, etc.;

[0102] 1. Initialize the graphical user interface (GUI) and build an interface for users to interact with;

[0103] 2. The user adjusts parameters such as reaction time T and flight distance according to actual experimental conditions;

[0104] 3. The user clicks the calculation button to trigger the subsequent calculation process;

[0105] 4. Perform a traversal operation on the reaction time corresponding to the diameter of the nozzle 140 and the pushing speed of the pushing component 170;

[0106] 5. Calculate the total time under each strategy;

[0107] 6. Select the best result from the calculated results and recommend it to the user;

[0108] 7. The method is executed and the process ends.

[0109] The workflow of the rapid freezing quenching device 10 of the above embodiment includes the following steps, for example:

[0110] 1. First, the reaction time to be observed is given, which should be within the range of 10ms~20s;

[0111] 2. Use software to calculate the required parameters of the electric push rod, such as linear speed, nozzle 140 specifications, and flight distance;

[0112] 3. According to the configuration information provided by the software, set the parameters of the rapid freezing quenching device 10;

[0113] 4. After the sample is loaded in the injection part, the pushing part 170 is operated to push the sample out, and the mixture is ejected at high speed through the nozzle 140 and is quickly frozen and formed when it contacts the surface of the freezing part 220;

[0114] 5. Finally, a small spatula made of polytetrafluoroethylene is used to collect the sample into a quartz tube to prepare for subsequent tests.

[0115] Furthermore, the present invention also provides a rapid freezing quenching method. The rapid freezing quenching method uses the rapid freezing quenching device of any of the above examples, and includes the following steps:

[0116] Samples are respectively injected into the first convection channel and the second convection channel of the mixing component. The samples are mixed and ejected through the confluence channel and the nozzle onto the freezing mechanism for freeze molding.

[0117] Specific embodiments are provided below to further illustrate the present invention. The present invention provides the following specific embodiments for better further understanding of the present invention, which are not limited to the specific implementation manners and do not constitute a limitation to the protection scope of the present invention.

[0118] Embodiment 1

[0119] This embodiment provides a rapid freezing quenching device 10, which includes a mixing and spraying mechanism 100 and a freezing mechanism 200.

[0120] The mixing and spraying mechanism 100 includes a first injection component 110, a second injection component 120, a mixing component 130, a nozzle 140, a first sample delivery pipeline 150, a second sample delivery pipeline 160, a pushing component 170, and a position adjusting component 180.

[0121] The mixing component 130 is provided with a first convection channel 131, a second convection channel 132, and a confluence channel 133. One end of the first convection channel 131 and one end of the second convection channel 132 are connected and converge at one end of the confluence channel 133. The first injection component 110 is communicated with the first convection channel 131 through the first sample delivery pipeline 150. The second injection component 120 is communicated with the second convection channel 132 through the second sample delivery pipeline 160. The nozzle 140 is connected to the mixing component 130 and communicated with the other end of the confluence channel 133. The pushing component 170 is used to push the first injection component 110 and the second injection component 120 for synchronous injection sampling.

[0122] Both the first convection channel 131 and the second convection channel 132 are linear channels. The included angle between the first convection channel 131 and the second convection channel 132 is 180°. The confluence channel 133 is a linear channel. The included angle between the confluence channel 133 and the first convection channel 131 and the second convection channel 132 is 90°. The lengths of the first convection channel 131, the second convection channel 132, and the confluence channel 133 are all 1 cm, and the inner diameters are all 1 mm.

[0123] The specifications of the first injection component 110 and the second injection component 120 are 1 mL. The pushing component 170 is a high-precision electric push rod with a maximum speed of 100 mm / s and a displacement accuracy of 0.01 mm.

[0124] The mixing and injection mechanism 100 includes three replaceable nozzles 140 with diameters of 0.10 mm, 0.15 mm, and 0.20 mm respectively.

[0125] The position adjustment component 180 includes a vertical adjustment member 181 and a horizontal adjustment member 182. The vertical adjustment member 181 is a manual displacement stage. The horizontal adjustment member 182 is a metal rod. The horizontal adjustment member 182 is connected to the vertical adjustment member 181 through an optical cross fixing clamp 183. The mixing component 130 is connected to the horizontal adjustment member 182.

[0126] The freezing mechanism 200 includes a cooling component 210, a receiving freezing component 220, a rotational drive component 230, a connecting rod 240, and a cover box 250.

[0127] The cooling component 210 includes a cooling medium container 211 and a cooling medium delivery pipe 212. The cooling medium delivery pipe 212 is connected to the side wall of the cooling medium container 211.

[0128] The receiving freezing component 220 is located below the nozzle 140. The receiving freezing component 220 is conical. The receiving freezing component 220 extends into the cooling medium container 211. An annular groove 221 is provided at the upper end of the receiving freezing component 220 and is arranged around the rotation axis. The rotational drive component 230 is located above the receiving freezing component 220 and is connected to the receiving freezing component 220 through the connecting rod 240. The mixing component 130, the nozzle 140, and the receiving freezing component 220 are all housed in the cover box 250.

[0129] When the nozzle with a diameter of 0.10 mm is used in the rapid freezing quenching device 10 of this embodiment, the flow rate of the sample, the velocity of the nozzle cross-section, and the quenching reaction time under different push rod speeds are shown in Table 1.

[0130] Table 1

[0131]

[0132] When the nozzle with a diameter of 0.15 mm is used, the flow rate of the sample, the velocity of the nozzle cross-section, and the quenching reaction time under different push rod speeds are shown in Table 2.

[0133] Table 2

[0134]

[0135] When using a nozzle with a diameter of 0.20 mm, the flow rate of the sample, the velocity at the nozzle cross-section, and the quenching reaction time at different push rod speeds are shown in Table 3.

[0136] Table 3

[0137]

[0138] The rapid freezing quenching device 10 of the above embodiments has the following technical effects:

[0139] 1. A mixing component 130 with a special internal structure is adopted. This mixing component 130 can make the mixing process of the sample extremely rapid, effectively control the dead volume at the same time, greatly reduce the dead time in the sample preparation process, improve the time resolution of the device, and can better capture the intermediates in the rapid reaction process, making up for the defects of too long dead time and large sample consumption in the traditional technology.

[0140] 2. A nozzle 140 with an extremely small diameter is connected below the mixing component 130. When the mixed liquid sprays out from the nozzle 140, it will obtain a very high initial velocity, significantly shortening the flight time. Moreover, through the cooperation of the relevant specifications and dimensions of the injection component, the conveying pipeline, the convection channel, and the confluence channel, and with the precise control of a high-precision electric push rod, the above device can achieve a time resolution of the order of microseconds.

[0141] 3. The specifications of the injection component and the nozzle 140 can be flexibly replaced according to needs, and the flight distance of the mixture can be adjusted by adjusting the relative distance between the nozzle 140 and the receiving freezing component 220. This enables the device to be applicable to the quenching operation of rapid reactions with reaction times between 5 milliseconds and 20 seconds, making up for the defect of a short monitoring time period in the traditional technology.

[0142] The above technical effects can significantly improve the application effect of the rapid freezing quenching technology in the electron paramagnetic resonance technology and enhance the detection ability of rapid reaction intermediates.

[0143] The technical features of the above embodiments can be combined arbitrarily. For the sake of concise description, not all possible combinations of the technical features in the above embodiments are described. However, as long as the combinations of these technical features do not conflict, they should all be considered as the scope recorded in this specification.

[0144] The above-described embodiments merely represent several implementation manners of the present invention. The description thereof is relatively specific and detailed, but it should not be construed as a limitation on the scope of the invention patent. It should be noted that for those of ordinary skill in the art, without departing from the concept of the present invention, several modifications and improvements can still be made, and these all fall within the protection scope of the present invention. Therefore, the protection scope of the present invention patent shall be subject to the appended claims, and the specification can be used to interpret the content of the claims.

Claims

1. A mixing injection mechanism of a rapid freezing and quenching device, characterized in that, It includes a mixing component and a nozzle; The mixing component is provided with a first convection channel, a second convection channel and a confluence channel. One end of the first convection channel and one end of the second convection channel are connected and converge at one end of the confluence channel. The other end of the first convection channel is used to communicate with a first injection component, the other end of the second convection channel is used to communicate with a second injection component, and the nozzle is communicated with the other end of the confluence channel.

2. The hybrid injection mechanism according to claim 1, characterized in that, The mixing and spraying mechanism also conforms to at least one of the following features (1) to (7): (1) Both the first convection channel and the second convection channel are straight channels, and the included angle is 150° to 180°; (2) The confluence channel is a straight channel, and the included angles between the confluence channel and the first convection channel and the second convection channel are 90° to 105°; (3) The length of the first convection channel is 0.5 cm to 1 cm, and the inner diameter is 0.5 mm to 1 mm; (4) The length of the second convection channel is 0.5 cm to 1 cm, and the inner diameter is 0.5 mm to 1 mm; (5) The length of the confluence channel is 0.5 cm to 1 cm, and the inner diameter is 0.5 mm to 1 mm; (6) The caliber of the nozzle is 0.1 mm to 0.2 mm; (7) The mixing and spraying mechanism includes a plurality of nozzles with different calibers, and each nozzle is detachably connected to the mixing component. Optionally, it includes nozzles with calibers of 0.1 mm, 0.15 mm and 0.2 mm respectively.

3. The hybrid injection mechanism according to claim 1 or 2, characterized in that, The mixing and spraying mechanism also includes a first injection component, a second injection component, a first sample delivery pipe and a second sample delivery pipe. The first injection component is communicated with the first convection channel through the first sample delivery pipe, and the second injection component is communicated with the second convection channel through the second sample delivery pipe.

4. The hybrid injection mechanism according to claim 3, characterized in that, The mixing and spraying mechanism also conforms to at least one of the following features (1) to (4): (1) The specification of the first injection component is 500 μL to 2 mL; (2) The specification of the second injection component is 500 μL to 2 mL; (3) The inner diameter of the first sample delivery pipe is 0.13 mm to 0.5 mm; (4) The inner diameter of the second sample delivery pipe is 0.13 mm to 0.5 mm.

5. The hybrid injection mechanism according to claim 3, characterized in that, The mixing and spraying mechanism also includes a pushing component, and the pushing component is used to push the first injection component and the second injection component to perform synchronous injection sampling.

6. The hybrid injection mechanism according to claim 5, characterized in that The mixing and spraying mechanism conforms to at least one of the following features (1) to (2): (1) The pushing speed of the pushing component can reach 100 mm / s, and the displacement accuracy is 0.01 mm to 0.02 mm; (2) The pushing component is an electric push rod.

7. A rapid freezing quenching device, characterized in that, It includes the mixing and spraying mechanism according to any one of claims 1 to 6 and a freezing mechanism. The freezing mechanism is used to receive the mixture ejected from the nozzle and freeze the mixture.

8. The rapid freezing and quenching device according to claim 7, characterized in that, The freezing mechanism includes a cooling component, a receiving freezing component, and a rotation driving component. The receiving freezing component is located below the nozzle for receiving the mixture ejected from the nozzle. The cooling component is used to cool the receiving freezing component, and the rotation driving component is connected to the receiving freezing component for driving the receiving freezing component to rotate.

9. The rapid freezing and quenching device according to claim 8, characterized in that, The rapid freezing and quenching device further conforms to at least one of the following features (1) to (3): (1) An annular groove is provided at the upper end of the receiving freezing component and is arranged around the rotation axis; (2) The cooling component includes a cooling medium container with an opening at the upper end. The receiving freezing component extends into the cooling medium container through the opening, and the size of the receiving freezing component gradually decreases from the upper end to the lower end; (3) The freezing mechanism further includes a housing box, and the mixing component, the nozzle, and the receiving freezing component are all housed in the housing box.

10. A rapid freezing quenching method, characterized in that, Using the rapid freezing and quenching device according to any one of claims 7 to 9, the rapid freezing and quenching method includes the following steps: Samples are respectively injected into the first convection channel and the second convection channel of the mixing component. The samples are mixed and ejected through the confluence channel and the nozzle onto the freezing mechanism for freeze molding.

Images