CN104117429B - A kind of heating oscillating magnetic flux separator - Google Patents

Abstract

The invention discloses a kind of heating oscillating magnetic flux separator, comprise three primary modules, i.e. Magneto separate module, heating module and vibration module.Magneto separate module includes linear electric motors, leading screw, magnet carrier, permanent magnet, and heating module includes electromagnetism load push-pull, spring, gripper shoe, support column, heating tank, heating film, and vibration module includes vibrating motor, eccentric hammer.This device drives magnet carrier to move on leading screw by linear electric motors, thus drives the up and down motion of permanent magnet, controls permanent magnet and realizes reunion and the dispersion of magnetic nanoparticle with the relative position of microwell plate；Promoting gripper shoe up and down motion by electromagnetism load push-pull, and then drive heating tank to move up and down by support column, the distance controlling heating tank and microwell plate hole wall realizes heating and the function closed.Finally achieve biological specimen processing means based on Magneto separate that is fully functional, small and exquisite and that be easily integrated, utilize robot arm device can realize the automation of whole experimentation.

Description

A heating and oscillating magnetic separation device

technical field

The invention relates to biological sample processing technology, in particular to a high-throughput fully automatic heating and oscillating magnetic separation device.

Background technique

Biological samples are diverse and include plasma, serum, whole blood, lymph, saliva, various tissues, hair, urine, bile, tears, spinal fluid, sweat, milk, amniotic fluid, feces, and exhaled air. The research on biological samples first requires the pretreatment of biological samples, which usually requires fast and efficient operation of a large number of samples at one time. Therefore, appropriate biological sample processing and analysis equipment can greatly reduce the workload of researchers and improve work efficiency. , to better promote the development of disciplines.

With the rapid development of nanoscience and technology, nanomaterials have been gradually applied to the field of life sciences, providing new technologies and means for the research and development of life sciences. Among them, the preparation and application of magnetic nanoparticles has become a research hotspot. Magnetic nanoparticles have the advantages of large specific surface area, superparamagnetism, and easy surface modification, and are widely used in the field of biomedicine. Among them, the use of magnetic nanoparticles for the separation of biological samples is one of its important applications in this field. When magnetic nanoparticles separate biomolecules, they have special physical and chemical properties and biocompatibility such as fast separation speed, high efficiency, reusability, simple operation, easy functionalization, easy automation, and no influence on the activity of the separated substances. features. Furthermore, the advantages of using magnetic nanoparticles in automatic detection are very prominent: traditional separation methods such as phenol-chloroform method, Chelex100 method, salting-out method, etc., have steps such as liquid-liquid separation and centrifugation, and the centrifuge equipment is bulky and cumbersome to operate , it takes a long time and is not suitable for the needs of automatic detection. Compared with the above methods, the separation operation of magnetic nanoparticles is simple and the required time is short. When an external magnetic field is applied, the magnetic nanoparticles are magnetized and adsorbed on the magnetic poles. When the magnetic field disappears, the magnetic properties of the magnetic nanoparticles are also simultaneously Disappeared and redispersed in the reaction solution. The use of magnetic nanoparticles for separation and positioning of biological samples is easier to achieve integration and automation.

Magnetic nanoparticles are widely used in cell separation, immunoassay, protein and enzyme immobilization, and DNA detection, etc. The biological sample processing method based on magnetic separation usually includes sample addition, cell lysis, DNA adsorption, magnetic separation, heating and mixing And other steps, the corresponding automation equipment needs to have the functions of adding samples, magnetic separation, heating and oscillation, including mechanical arms, static magnetic separation devices, heating devices, oscillating mixing devices and other functional devices. Each device corresponds to at least one station. To complete the process of biological sample processing, the robotic arm needs to repeatedly complete the transfer of the microwell plate between the above-mentioned multiple stations, which greatly increases the workload and brings great difficulties to the experiment. A lot of inconvenience, while greatly increasing the possibility of cross-contamination. Further, this type of instrument is not conducive to small modularization and integration into a large integrated automation workstation.

Contents of the invention

Technical problem: Aiming at the deficiencies of the prior art, the present invention provides a fully automatic heating and oscillating magnetic separation device, which integrates the magnetic separation device, temperature control device and oscillating mixing device to provide a microplate bracket and a permanent The magnet, heating mechanism, and oscillating mechanism can conveniently realize the functions of magnetic separation, heating or oscillating, and have the advantages of completeness, small size, and convenient integration. The processing of biological samples can be completed without moving the microplate during the experiment, which solves the defects of cumbersome operation, high probability of cross-contamination, and poor controllability caused by repeated transfer of microplates, as well as the large volume integrated into the automated workstation. shortcoming.

Technical solution: For the above technical problems, the present invention provides a heating and oscillating magnetic separation device, which includes three main modules, namely a magnetic separation module, a heating module and a vibration module, and an auxiliary mechanism;

The magnetic separation module includes linear motors, lead screws, magnet brackets, and permanent magnets; the heating module includes electromagnetic pushers, support plates, springs, support columns, heating tanks, and heating films; the vibration module includes vibration motors and eccentric hammers; the auxiliary mechanism includes Base, fixed plate, 4 elastic columns, main frame;

The base is in the shape of a groove, and there are three fixing grooves in the middle, which are respectively used to fix the linear motor, the electromagnetic push-pull and the vibration motor, and two fixing plates are placed on both sides of the middle; four elastic pillars are set at the four corners of the fixing plate A fixing groove, the elastic column is placed vertically in the fixing groove;

The main body frame is placed on four elastic pillars, an eccentric hammer is connected to the lower end of the center of the main body frame, and the eccentric hammer is connected to the vibration motor;

There are two fixed terminals on both sides of the short side of the top of the main frame, and the fixed terminals have built-in balls for fixing the micropore plate, so that the micropore plate is always in the same position; the fixed terminals are respectively located on the two sides of the micropore plate The short side does not affect the gripping of the microplate by the robotic arm;

One end of the lead screw is fixed on the linear motor and placed vertically. The lead screw is connected with a magnet bracket, and the magnet bracket extends outwards to a rectangular parallelepiped bracket. The bracket is provided with 4 magnetic hole holes for placing 4 magnet brackets. Permanent magnets, the permanent magnets are rectangular parallelepiped, and the permanent magnets are placed in parallel, and the adjacent tube arrays on the microwell plate correspond to one of the four permanent magnets, so as to realize the agglomeration of magnetic nanoparticles on the side wall;

A support plate is fixed on the electromagnetic push-pull, and 2 support columns are fixed on both sides of the support plate, and the 2 support columns are placed in parallel on the same horizontal line. Fixed heating tank at the top of the column;

The heating tank is an irregular cuboid, and the outer sides of its two long sides are smooth heating surfaces, and a heating film is placed respectively; the heating tank has 8 contact surfaces, corresponding to each row in the micro-orifice tube array, The contact surface is a structure of multiple conical surfaces. The taper of the conical surface matches the taper of the tubular container to achieve seamless fit with the hole wall; the top of the heating tank is a via hole that is suitable for the microplate, making the microplate It can be placed in the heating tank to fit the contact surface; the small holes on the top of the heating tank correspond to the small holes on the microporous plate, which can accelerate heat conduction.

Preferably, the contact surface in the heating tank and the permanent magnet are located in different tube arrays of microplates, and the two do not collide.

Beneficial effect:

(1) The present invention overcomes the deficiency of a single device and a single function in the existing biological sample processing automation equipment based on magnetic separation, and integrates the magnetic separation device, heating device and vibration device into one device to fully ensure the concentration of the solution in the microwell plate. Magnetic separation, thorough mixing and heating incubation at a certain temperature ensure the integrity of the function.

(2) In the whole working process of the present invention, the microporous plate containing the reaction solution is fixed statically above the main body frame all the time, and the single device realizes the operation of magnetic separation, heating and vibration, which saves the working space and makes the device small and exquisite, ensuring The effect of biological sample processing is improved, and the interference between microporous tubes can be avoided, which greatly prevents cross-contamination;

(3) Modular design, the magnetic separation, heating and vibration modules in this device can be used independently without interfering with each other;

(4) Strong practicability. The device is an open structure with the characteristics of simple mechanical structure, ingenious conception, and easy realization. It can be integrated into large-scale biochemical analysis instruments and other devices, and can also be used independently as a miniaturized biological sample processing device. In use, such devices meet the actual needs of users.

Description of drawings

Fig. 1 is the overall outline drawing of the present invention;

Fig. 2 is a component front view of the present invention;

Fig. 3 is a component rear view of the present invention;

Fig. 4 is the bottom schematic diagram of the present invention;

Fig. 5 is a schematic diagram of a magnet bracket of the present invention;

Fig. 6 is a schematic diagram of a heating tank of the present invention;

In the figure: base 101, electromagnetic push-pull 102, fixed plate 103, elastic pillar 104, main frame 105, ball 106, fixed terminal 107, heating tank 108, heating film 109, vibration motor 201, linear motor 202, lead screw 203, Magnet bracket 204 , permanent magnet 205 , support plate 206 , support column 208 , eccentric hammer 301 , heating surface 501 , contact surface 502 , via hole 503 , and small hole 504 .

detailed description

Below in conjunction with accompanying drawing and embodiment mode, the patent of the present invention is described in further detail:

A heating and oscillating magnetic separation device includes three main modules, namely a magnetic separation module, a heating module and a vibration module. The magnetic separation module includes linear motors, lead screws, magnet brackets, and permanent magnets. The heating module includes electromagnetic pushers, springs, support plates, support columns, heating tanks, and heating films. The vibration module includes vibration motors and eccentric hammers. In addition, it also includes auxiliary structures such as bases, fixed plates, elastic columns and main frames.

The base is in the shape of a groove, and the middle part is fixed with a linear motor, an electromagnetic push-pull and a vibrating motor, and two fixing plates are placed on both sides of the middle.

The four corners of the fixing plate are provided with four elastic pillar fixing grooves, and the elastic pillars are placed vertically in the fixing grooves;

The main body frame is placed on four elastic pillars, and an eccentric hammer is connected to the lower end of the center of the main body frame, and the eccentric hammer is connected to the vibration motor; there are two fixed terminals on both sides of the short side of the top of the main frame, and the fixed terminals are built-in Roller balls are used to fix the microplate so that the microplate is always in the same position. The fixed terminals are respectively located on the two short sides of the micro-orifice plate, so as not to affect the gripping of the micro-orifice plate by the mechanical arm;

One end of the lead screw is fixed on the linear motor and placed vertically. The lead screw is connected with a magnet bracket. The magnet bracket is provided with 4 permanent magnet slots for placing 4 permanent magnets. The permanent magnets are rectangular parallelepiped , the permanent magnets are placed in parallel, and a permanent magnet can be placed between adjacent tube arrays on the microwell plate;

A support plate is fixed on the electromagnetic push-pull, and 2 support columns are fixed on both sides of the support plate, and the 2 support columns are placed in parallel on the same horizontal line. A heating bath is fixed on the top of the column. The heating tank is an irregular cuboid, and the outer sides of the two long sides are smooth heating surfaces, and a heating film is placed respectively; the heating tank has 8 contact surfaces, corresponding to each row in the microplate array one by one, and the contact surfaces It is a structure of multiple tapered surfaces, and the taper of the tapered surface matches the taper of the tubular container to achieve seamless fit with the hole wall. The top of the heating tank is a via hole adapted to the microplate, so that the microplate can be placed in the heating tank and bonded to the contact surface; the small holes on the top of the heating tank correspond to the small holes on the microplate, which can accelerate heat conduction.

The device of the present invention drives the magnet bracket to move on the lead screw through a linear motor, thereby driving the permanent magnet to move up and down, and controls the relative position between the permanent magnet and the hole wall of the micropore plate to realize the agglomeration and dispersion of magnetic nanoparticles; through electromagnetic push-pull The device pushes the support plate to move up and down, and then drives the heating tank to move up and down through the support column, and controls the distance between the heating tank and the hole wall of the microhole plate to realize the function of heating and closing; the vibration motor controls the eccentric hammer to drive the main frame to realize the vibration function of the whole device . Finally, the automation of the entire nucleic acid extraction can be completed by controlling the different timings of magnetic separation, vibration, and heating.

Further, in the present invention, after the electromagnetic push-pull device is energized, a magnetic field is generated to drive the support plate to move upwards, so that the heating tank and the hole wall of the micropore plate are in harmony, and the heating tank is made of a metal material (gold, silver, aluminum, iron, etc.), The heating film on both sides is energized, and the principle of metal heat conduction is used to realize the temperature conduction of the heating tank and realize the heating function of the microporous plate. After the electromagnetic push-pull is powered off, the magnetic field disappears, driving the support plate to move downward, generally 3mm, the heating tank is separated from the microporous plate, the heating film is powered off, and the heating function is turned off.

Further, in the present invention, the spring of the heating module plays the role of supporting and buffering the heating tank, preventing violent vibration when the electromagnetic push-pull moves up and down.

Further, in the present invention, the elastic pillars play the role of fixation and buffering. After the vibration motor is energized, the eccentric hammer is driven to drive the main frame to vibrate back and forth, thereby driving the vibration of the micro-orifice plate fixed on the fixed terminal at the top of the main frame, and the vibration stops after the vibration motor is turned off.

Further, in the present invention, the linear motor drives the lead screw to drive the magnet bracket to move upward, so that the 4 permanent magnets on the magnet bracket are located between the walls of every two rows of the micro-orifice plate array to realize the reunion of magnetic nanoparticles; The magnet bracket moves downward, and the 4 permanent magnets are located at the lower end of the microwell plate array, which lose the adsorption effect on the magnetic nanoparticles.

Further, in the present invention, the contact surface in the heating tank and the permanent magnet are in different arrays of microplates, and the two do not collide.

Further, in the present invention, the control sequence of the magnetic field, heating and vibration functions has the following six types: 1) no magnetic field, no heating, no vibration; 2) no magnetic field, heating, and vibration; 3) magnetic field, no heating , No vibration; 4) No magnetic field, no heating, but vibration; 5) Magnetic field, no heating, no vibration; 6) No magnetic field, heating, no vibration.

As shown in Fig. 1 to Fig. 5, a kind of heating vibration magnetic separation device of the present invention comprises base 101, and described base 101 is an irregular groove, is provided with three fixed grooves in the middle, fixes linear motor 202, electromagnetic push-pull 102 and the vibrating motor 201, the two sides in the middle are protruding cuboids, and two fixing plates 103 are placed. The four corners of the fixing plate 103 are fixed with four elastic pillars 104, and the elastic pillars 104 are cylindrical and placed vertically in the fixing groove, in order to prevent damage to the instrument during vibration and play a buffering role. The top of the elastic column 104 is provided with a fixing slot for fixing and supporting the main body frame 105 . The ball 106 is fixed on the fixed terminal 107, and the fixed terminal 107 is located on both sides of the top of the main body frame 105. The fixed terminal 107 is built with the ball 106 for fixing the microporous plate, so that the microporous plate is always in the same position. The fixed terminals 107 are respectively located on the two short sides of the microporous plate, so as not to affect the grasping of the microporous plate by the mechanical gripper. The lower end of the center of the main frame 105 is connected to an eccentric hammer 301, and the eccentric hammer 301 is connected to the vibration motor 201. After the vibration motor 201 is powered on, the eccentric hammer 301 is driven to drive the main frame 105 to realize reciprocating vibration, thereby driving the fixed terminal 107 fixed on the top of the main frame 105 Vibration of the microplate. One end of the lead screw 203 is fixed on the linear motor 202 and placed vertically. The lead screw 203 is connected with a magnet bracket 204, and the magnet bracket 204 extends outward a rectangular parallelepiped support, and the support is provided with 4 magnetic block holes. , place four permanent magnets 205, the permanent magnets 205 are cuboid, and the permanent magnets 205 are placed in parallel. The common microplates on the market are divided into 6 holes, 12 holes, 24 holes, 48 holes, 96 holes, etc. according to different models. 384 wells and 1536 wells, etc., the wells on the microwell plate are arranged in an orderly array, and each well is equipped with a tubular container, which forms a tube array. The intervals between columns are also parallel to each other, therefore, each permanent magnet 205 is stuck in the interval of two tube arrays, realizing the agglomeration and dispersion of the magnetic particles in the tubular containers on both sides on the side walls. A support plate 206 is fixed on the electromagnetic push-pull 102, two support columns 208 are fixed on both sides of the support plate 206, and the two support columns 208 are placed in parallel on the same horizontal line, and the spring 207 is located at the lower end of the support plate 206. Between the two support columns 208 , the top of the support column 208 fixes the heating tank 108 . The heating tank 108 is an irregular cuboid, with smooth heating surfaces 501 on the outside of both sides, and a heating film 109 is respectively placed; the heating tank 108 has eight contact surfaces 502, corresponding to each row in the microplate array , the contact surface 502 is a structure of multiple tapered surfaces, and the taper of the tapered surface matches the taper of the tubular container to achieve seamless fit with the hole wall. The top of the heating tank 108 is a via hole 503 adapted to the micro-orifice plate, so that the micro-orifice plate can be placed in the heating tank 108 and bonded to the contact surface 502; the small hole 504 on the top of the heating tank 108 corresponds to the small hole on the micro-orifice plate, Accelerates heat conduction.

Taking the nucleic acid extraction process based on magnetic separation as an example, the specific working process of this device is as follows:

1. At the beginning of the work, the microplate is stuck between the two balls 106, placed stably, and the microplate contains a solution mixed with nucleic acid extraction samples and lysate. At this time, the device is in an initialization state, and both the magnet bracket 204 and the support plate 206 are located at the lowest points in their respective vertical directions. The magnetic field of the permanent magnet 205 has no effect on the solution in the micro-orifice plate, the heating tank 108 is separated from the tube wall in the micro-orifice plate array, and the electromagnetic push-pull 102, vibration motor 201 and linear motor 202 are all in a non-working state.

2. Timed incubation. Power the electromagnetic push-pull device 102 to generate a magnetic field, and drive the support plate 206 to move upwards, so that the heating tank 108 is in contact with the wall of the micropore plate, and then the heating film 109 on both sides is energized, and the temperature of the heating tank 108 is realized by using the principle of metal heat conduction. Conduction to realize the heating function of the microplate. After a certain period of time, the electromagnetic push-pull device is powered off, and the magnetic field disappears at this time, driving the support plate to move down about 3 mm, the heating tank is separated from the microporous plate, the heating film 109 is powered off, and the heating function is turned off.

3. Shake to mix. Choose whether to enable the vibration function according to the experimental requirements. While heating, the vibration motor 201 can be turned on to drive the eccentric hammer 301 to drive the main frame 105 to vibrate, so as to achieve uniform mixing of the solution in the microwell plate. Turn off the vibration motor while turning off the heat.

4. Magnetic separation. Add the magnetic particle solution specifically combined with the nucleic acid extraction sample into the microwell plate, turn on the linear motor 202 to drive the lead screw 203 to drive the magnet bracket 204 to move upward, so that the permanent magnet 205 stops on the side wall of the microwell plate, and can simultaneously Adsorb the solutions in the arrays on both sides. At this time, the magnetic particles in the solution carry the combined substances to be separated and are adsorbed to the side walls, realizing the reunion of the side walls. The remaining liquid can be sucked away by means of pipettes, etc. Separation of non-target substances.

5. Release magnetic nanoparticles. The linear motor 202 reversely drives the lead screw 203 to drive the magnet bracket 204 to move downwards, so that the permanent magnet 205 stops at the bottom of the microporous plate. At this time, the magnetism of the permanent magnet 205 loses its adsorption effect on the solution in the microporous plate.

6. Oscillating and mixing, adding cleaning solution into the microporous plate, turning on the vibrating motor 201, driving the eccentric hammer 301 to drive the main body frame 105 to vibrate, so as to realize the re-mixing of the magnetic nanoparticle solution in the microporous plate. Turn off the vibration motor.

7. Repeat steps 4-6 several times to complete the washing, and add the eluent to the microwell plate.

8. Magnetic separation. Turn on the linear motor 202 to drive the lead screw 203 to drive the magnet bracket 204 to move upward, The permanent magnet 205 is docked on the side wall of the tube hole of the micro-orifice plate, which can simultaneously adsorb the solutions in the arrays on both sides. At this time, the magnetic particles in the solution carry the combined substances to be separated and are adsorbed to the side wall, realizing the reunion of the side wall, which can be used Transfer the remaining liquid by means of a pipette or the like, and the transferred liquid is the extracted substance.

The conversion of the above-mentioned several working states can be realized through the control system, pay attention to the timing control, and combine with the pipetting device to realize the automatic biological sample processing experiment.

The above is only a preferred embodiment of the present invention, it should be pointed out that for those of ordinary skill in the art, without departing from the principle of the present invention, some improvements and modifications can also be made. It should be regarded as the protection scope of the present invention.

Claims (2)

1. a heating oscillating magnetic flux separator, it is characterised in that: this device includes three primary modules, i.e. magnetic Separation module, heating module and vibration module, and auxiliary body；

Magneto separate module includes linear electric motors (202), leading screw (203), magnet carrier (204), permanent magnet (205)； Heating module include electromagnetism load push-pull (102), gripper shoe (206), spring (207), support column (208), Heating tank (108), heating film (109)；Vibration module includes vibrating motor (201), eccentric hammer (301)； Auxiliary body comprises base (101), fixed plate (103), 4 elastic pillars (104), main frames (105)；

Described base (101) is in groove-like, and centre is provided with three fixing grooves, is respectively used to fixing linear electric motors (202), electromagnetism load push-pull (102) and vibrating motor (201), middle two lateral processes two pieces of fixed plates of placement (103)；Described fixed plate (103) corner is provided with 4 elastic pillars and fixes groove, described elastic pillar (104) In vertically placing in fixing groove；

Described main frame (105) is placed on 4 elastic pillars (104), in described main frame (105) Heart lower end connects an eccentric hammer (301), and eccentric hammer (301) is connected with vibrating motor (201)；

Minor face both sides, described main frame (105) top are two fixing terminals (107), described fixing terminal (107) Built-in ball (106), is used for fixing microwell plate so that microwell plate is in same position all the time；Described fixing end Son (107) lays respectively at two minor faces of microwell plate, thus does not affect the mechanical arm crawl to microwell plate；

Described leading screw (203) one end is fixed on linear electric motors (202), in vertically placing, on described leading screw Connecting magnet carrier (204), magnet carrier (204) stretches out the support of a cuboid, and support sets Have 4 magnetic patch holes, place 4 pieces of permanent magnets (205), permanent magnet (205) in cuboid, permanent magnet (205) Between be placed in parallel, between pipe array adjacent on microwell plate respectively in corresponding 4 pieces of permanent magnets (205) one Individual, it is achieved magnetic nanoparticle is in the reunion of sidewall；

Fixing a gripper shoe (206) on described electromagnetism load push-pull (102), the upper both sides of gripper shoe (206) are solid Fixed 2 support columns (208), 2 support columns (208) are positioned on same level line and are placed in parallel, described bullet Spring (207) is positioned at lower end in the middle of gripper shoe (206), is in the middle of two support columns (208), support column (208) heating tank (108) is fixed on top；

Described heating tank (108) is an irregular cuboid, is smooth heating outside two long limit Face (501), places heating film (109) respectively；Heating tank has 8 contact surfaces (502), one_to_one corresponding Every a line in microwell plate pipe array, contact surface (502) is the structure of multiple taper surface, the tapering of taper surface Mate with the tapering of tubular container, it is achieved seamless applying with hole wall；Heating tank top is the mistake of adaptive microwell plate Hole (503), makes microwell plate can put in heating tank and fits with contact surface (502)；Heating tank top little Aperture on the corresponding microwell plate in hole (504), can accelerate heat transfer.

Heating oscillating magnetic flux separator the most according to claim 1, it is characterised in that: described heating tank (108) contact surface (502) in is in the different pipe arrays into microwell plate from described permanent magnet (205), Both do not collide.