CN107629952B - Nucleic acid hybridization instrument - Google Patents
Nucleic acid hybridization instrument Download PDFInfo
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- CN107629952B CN107629952B CN201710843185.6A CN201710843185A CN107629952B CN 107629952 B CN107629952 B CN 107629952B CN 201710843185 A CN201710843185 A CN 201710843185A CN 107629952 B CN107629952 B CN 107629952B
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Abstract
The invention discloses a nucleic acid hybridization instrument which comprises a machine table, a pipetting mechanism, a sample washing mechanism, a liquid pumping mechanism and a control mechanism. The nucleic acid hybridization instrument automatically controls the sample adding driving device, the sample washing driving device and the liquid pumping mechanism to act through the control mechanism, can automatically add reagents, automatically incubate and automatically clean, does not need manual intervention in the whole incubation process, can automatically complete the incubation process, saves time and labor, is favorable for ensuring the consistency of the detection process, and is further favorable for realizing the standardized operation, the repeatability and the high flux of the nucleic acid in-situ hybridization experiment.
Description
Technical Field
The invention relates to the field of molecular biology detection, in particular to a nucleic acid hybridization instrument.
Background
RNA in situ nucleic acid hybridization (RNA nucleic acid hybridization in situ) is also known as RNA in situ hybridization histochemistry (RNA in situ hybridization histochemistry) or RNA in situ hybridization (RNA in situ hybridization RISH). The technology is an in situ hybridization technology for detecting RNA expression in cells and tissues by using probes such as cRNA or oligonucleotides. The basic principle is as follows: under the condition that the cell or tissue structure is kept unchanged, the labeled known RNA nucleotide fragment is combined with the corresponding gene fragment in the cell or tissue to be detected according to the base pairing principle, and the formed hybrid is observed under an optical microscope or an electron microscope after the color reaction. The RNA in situ hybridization technology is continuously improved, and the application field of the RNA in situ hybridization technology is far ahead of that of the DNA in situ hybridization technology. It is one of the most effective molecular pathology techniques, and can be used for qualitative, positional and quantitative analysis in gene analysis and diagnosis, and also can be used for effectively analyzing low abundance and rare mRNA expression.
Although the RNA in situ hybridization technology is a commonly used molecular detection technology, the technology has complex steps, is time-consuming and labor-consuming, comprises a plurality of simple and repeated cleaning and incubation processes, and has the problems of boring and complicated manual operation, time consumption, labor consumption and the like, and the consistency of all detection processes is difficult to ensure, so that the accuracy and the reliability of the results are often influenced.
Disclosure of Invention
Based on this, it is necessary to provide a nucleic acid hybridization apparatus to solve the problems that the conventional nucleic acid hybridization operation such as RNA is time-consuming and labor-consuming and it is difficult to ensure the consistency of the detection process.
The technical scheme for solving the technical problems is as follows.
A nucleic acid hybridization instrument, comprising:
the machine comprises a reagent zone, a needle washing zone, an incubation zone and a washing liquid zone, wherein the reagent zone is used for placing a reagent, the needle washing zone is provided with a needle washing pool, the incubation zone is provided with an incubation groove used for placing a reaction container, and the washing liquid zone is used for placing a washing liquid;
the sample adding driving device is arranged on the machine table and connected with the sample adding needle to drive the sample adding needle to move among the reagent zone, the needle washing zone and the incubation zone;
the sample washing mechanism comprises a liquid injection needle, a liquid suction needle and a sample washing driving device, wherein the sample washing driving device is arranged on the machine table and is respectively connected with the liquid injection needle and the liquid suction needle to be used for driving the liquid injection needle and the liquid suction needle to move between the needle washing area and the incubation area;
the liquid pumping mechanism is respectively connected with the sample adding needle, the needle washing pool, the liquid injection needle and the liquid suction needle; and
the control mechanism is connected with the sample adding driving device, the sample washing driving device and the liquid pumping mechanism;
the control mechanism is used for controlling the sample adding driving device to move the sample adding needle to the reagent area and controlling the liquid pumping mechanism to pump reagent from the reagent area through the sample adding needle; the control mechanism is also used for controlling the sample adding driving device to move the sample adding needle to the incubation area and controlling the liquid pumping mechanism to pump out the reagent in the sample adding needle to the reaction container in the incubation groove; the control mechanism is also used for controlling the sample adding driving device to move the sample adding needle into the needle washing pool and controlling the liquid pumping mechanism to pump the cleaning liquid from the cleaning liquid in the cleaning liquid area so as to clean the inner wall and the outer wall of the sample adding needle respectively; the control mechanism is also used for controlling the sample adding driving device to reset the sample adding needle; the control mechanism is also used for controlling the liquid pumping mechanism to pump out the cleaning waste liquid in the needle washing pool; the control mechanism is also used for controlling the sample washing driving device to move the liquid injection needle to the incubation area and controlling the liquid pumping mechanism to pump the cleaning liquid from the cleaning liquid in the cleaning liquid area and pump the cleaning liquid to the reaction container through the liquid injection needle; the control mechanism is also used for controlling the sample washing driving device to move the liquid sucking needle to the incubation area and controlling the liquid pumping mechanism to pump the washing waste liquid in the reaction container out through the liquid sucking needle; the control mechanism is also used for controlling the sample washing driving device to respectively move the liquid injection needle and the liquid suction needle into the needle washing pool, and controlling the liquid pumping mechanism to pump cleaning liquid from the cleaning liquid in the washing liquid area so as to respectively clean the liquid injection needle and the liquid suction needle; the control mechanism is also used for controlling the sample washing driving device to drive the liquid injection needle and the liquid suction needle to reset.
In one embodiment, the reagent zone has a reagent rack; the bottom of the reagent rack is provided with a first heating device, and the first heating device is connected with the control mechanism.
In one embodiment, a second heating device is arranged at the bottom of the incubation groove, and the second heating device is connected with the control mechanism; and/or
The bottom of the incubation groove is provided with a jacking mechanism, and the jacking mechanism is connected with the control mechanism so as to control one side of the reaction container in the incubation groove to be jacked by the control mechanism to form an inclination; and/or
A temperature sensor is arranged in the incubation groove and is electrically connected with the control mechanism; and/or
The top of the incubation groove is provided with a box cover.
In one embodiment, the nucleic acid hybridization instrument further comprises a box cover driving device connected with the control mechanism, wherein the box cover driving device is connected with the box cover and used for opening or closing the incubation groove by driving the box cover to move.
In one embodiment, the washing liquid area is provided with a plurality of washing liquid bottles, and the washing liquid bottles are respectively connected with the liquid pumping mechanism through pipelines; and each washing liquid bottle is provided with an independent control valve on a corresponding pipeline, and the control valve is connected with the control mechanism so as to be controlled to be opened or closed by the control mechanism.
In one embodiment, the liquid injection needles and the liquid suction needles are respectively arranged in rows, and the plurality of liquid injection needles in the rows are arranged at intervals from the plurality of liquid suction needles in the rows.
In one embodiment, the liquid pumping mechanism comprises a plunger pump and a first diaphragm pump, wherein the plunger pump and the first diaphragm pump are both connected with the sample adding needle in series, the plunger pump is used for controlling the sample adding needle to pump reagent from the reagent area and controlling the sample adding needle to pump the pumped reagent into the reaction container, and the first diaphragm pump is used for pumping cleaning liquid from the cleaning liquid area into the sample adding needle and pumping the cleaning liquid into the needle washing pool so as to clean the inner wall and the outer wall of the sample adding needle.
In one embodiment, the liquid pumping mechanism further comprises a second diaphragm pump, wherein the second diaphragm pump is connected in series with the needle washing tank and is used for pumping out waste liquid of the needle washing tank.
In one embodiment, the liquid pumping mechanism further comprises a third diaphragm pump and a fourth diaphragm pump, wherein the third diaphragm pump is connected with the liquid injection needle in series to pump the cleaning liquid from the cleaning liquid area and pump the cleaning liquid through the liquid injection needle, and the fourth diaphragm pump is connected with the liquid suction needle in series to pump the cleaning waste liquid from the reaction container through the liquid suction needle and pump the cleaning waste liquid.
In one embodiment, the nucleic acid hybridization apparatus further comprises a waste liquid bottle interface provided on the machine, and the waste liquid bottle interface is connected with the second diaphragm pump and the fourth diaphragm pump.
The nucleic acid hybridization instrument automatically controls the sample adding driving device, the sample washing driving device and the liquid pumping mechanism to act through the control mechanism, can automatically add reagents, automatically incubate and automatically clean, does not need manual intervention in the whole incubation process, can automatically complete the incubation process, saves time and labor, is favorable for ensuring the consistency of the detection process, and is further favorable for realizing the standardized operation, the repeatability and the high flux of the nucleic acid in-situ hybridization experiment.
Drawings
FIG. 1 is a schematic diagram showing the structure of an incubation slot of a nucleic acid hybridization apparatus according to an embodiment in an open state;
FIG. 2 is a schematic diagram showing the structure of the nucleic acid hybridization apparatus shown in FIG. 1 in a closed state of an incubation slot;
FIG. 3 is a top view of the nucleic acid hybridization instrument shown in FIG. 2;
FIG. 4 is a schematic diagram of the sample loading driving device in FIG. 1;
FIG. 5 is a schematic diagram of the sample washing driving device in FIG. 1;
FIG. 6 is a schematic structural view of a part of the structure of the nucleic acid hybridization instrument shown in FIG. 1;
FIG. 7 is a schematic view of the jack-up mechanism of FIG. 1;
FIG. 8 is a schematic diagram illustrating the cooperation between the jack-up mechanism shown in FIG. 7 and a portion of the structure shown in FIG. 6;
FIG. 9 is a schematic diagram of the whole machine of the nucleic acid hybridization instrument shown in FIG. 1;
FIG. 10 is a schematic view of a liquid path system mechanism of the nucleic acid hybridization instrument shown in FIG. 1;
fig. 11 is a schematic view of the structure of the filling needle, the pipetting needle and the hub in fig. 1;
fig. 12 is a schematic view of the injection needle, the aspiration needle and the needle holder shown in fig. 11 from another view.
Detailed Description
In order that the invention may be readily understood, a more complete description of the invention will be rendered by reference to the appended drawings. Preferred embodiments of the present invention are shown in the drawings. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.
It will be understood that when an element is referred to as being "fixed to" another element, it can be directly on the other element or intervening elements may also be present. When an element is referred to as being "connected" to another element, it can be directly connected to the other element or intervening elements may also be present.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and/or" as used herein includes any and all combinations of one or more of the associated listed items.
Referring to fig. 1, 2 and 3, a nucleic acid hybridization apparatus 10 according to an embodiment includes a machine 100, a pipetting mechanism 200, a washing mechanism 300, a pumping mechanism 400 and a control mechanism 500.
The station 100 has a reagent zone 102, a wash zone 104, an incubation zone 106, and a wash zone 108. The reagent zone 102 is used for placing reagents. The needle wash zone 104 has a needle wash basin 110. Incubation zone 106 has an incubation slot 120 for placing reaction vessel 20. The wash zone 108 is used to place a wash liquid.
The pipetting mechanism 200 includes a loading needle 210 and a loading drive device 220. The loading driving device 220 is disposed on the machine 100 and connected to the loading needle 210 for driving the loading needle 210 to move among the reagent area 102, the washing area 104 and the incubation area 106.
In one embodiment, the loading drive 220 includes a loading rail, a loading slide, and a loading drive motor. The sample adding guide rail, the sample adding sliding block and the sample adding driving motor form a sample adding arm group. The loading needle 210 can be moved forward, backward, left, right, up and down (i.e., moved randomly in three dimensions) by the loading arm set. The sample feeding driving motor is preferably a stepping motor, and the control precision is high.
Specifically, referring to fig. 4, in the embodiment shown in fig. 4, the loading rails include an X-axis loading rail 221, a Y-axis loading rail 222, and a Z-axis loading rail 223; the loading slide blocks comprise an X-axis loading slide block 224, a Y-axis loading slide block 225 and a Z-axis loading slide block 226; accordingly, the loading drive motors include an X-axis loading motor 227, a Y-axis loading motor 228, and a Z-axis loading motor 229. The Y-axis sample feeding guide rail 222, the Y-axis sample feeding slide block 225 and the Y-axis sample feeding motor 228 are integrally arranged on the X-axis sample feeding slide block 224, and the Y-axis sample feeding guide rail 222, the Y-axis sample feeding slide block 225 and the Y-axis sample feeding motor 228 can be driven to integrally move along the X-axis through the action of the X-axis sample feeding motor 227; the Z-axis sample feeding guide rail 223, the Z-axis sample feeding slide block 226 and the Z-axis sample feeding motor 229 are integrally arranged on the Y-axis sample feeding slide block 225, and the Z-axis sample feeding guide rail 223, the Z-axis sample feeding slide block 226 and the Z-axis sample feeding motor 229 can be driven to integrally move along the Y-axis through the action of the Y-axis sample feeding motor 228; the loading needle 210 is provided on the Z-axis loading slider 226, and can move up and down in the Z-axis by the action of the Z-axis loading motor 229.
More specifically, in the embodiment shown in fig. 4, the loading sliders (such as the X-axis loading slider 224, the Y-axis loading slider 225, and the Z-axis loading slider 226) are connected to the loading driving motors (such as the X-axis loading motor 227, the Y-axis loading motor 228, and the Z-axis loading motor 229) by a belt, and the loading sliders are fixed on one side of the belt, and the belt is driven to rotate by the rotation of the loading driving motors, so as to drive the loading sliders to move.
The sample washing mechanism 300 includes a liquid injection needle 310, a liquid suction needle 320, and a sample washing drive device. The sample washing driving device is disposed on the machine 100 and connected to the liquid injection needle 310 and the liquid suction needle 320, respectively, for driving the liquid injection needle 310 and the liquid suction needle 320 to move to the incubation area 106.
In one embodiment, the sample washing driving device comprises a sample washing guide rail, a sample washing sliding block, a sample washing driving motor and the like. The sample washing guide rail and the sample washing driving motor form a sample washing arm group. The priming needle 310 and the pipetting needle 320 can be driven by the sample washing arm set to move forward and backward and up and down. The sample washing driving motor preferably adopts a stepping motor, and has high control precision.
Specifically, referring to FIG. 5, in the embodiment shown in FIG. 5, the wash rails include a Y-axis wash rail 331 and a Z-axis wash rail 332; the sample washing slide blocks comprise a Y-axis sample washing slide block 333 and a Z-axis sample washing slide block 334; the sample washing drive motor includes a Y-axis sample washing motor 335 and a Z-axis sample washing motor 336. The Z-axis sample washing rail 332, the Z-axis sample washing slider 334, and the Z-axis sample washing motor 336 are provided on the Y-axis sample washing slider 333, and can be driven by the Y-axis sample washing motor 335 to move forward and backward along the Y-axis sample washing rail 331. The injection needle 310 and the pipetting needle 320 are provided on the Z-axis sample washing slider 334, and can be driven by the Z-axis sample washing motor 336 to move up and down along the Z-axis sample washing rail 332.
Further, in the embodiment shown in fig. 5, the Y-axis sample washing slide block 333 is in transmission connection with the Y-axis sample washing motor 335 through a transmission belt, the Y-axis sample washing slide block 333 is fixed at one side of the transmission belt, and the rotation of the Y-axis sample washing motor 335 can drive the transmission belt to rotate so as to drive the Y-axis sample washing slide block 333 to move. The Z-axis sample washing motor 336 is connected with the Z-axis sample washing slide block 334 through a screw rod transmission assembly, and drives the Z-axis sample washing slide block 334 to move up and down through a screw rod transmission principle.
It will be appreciated that in other embodiments, the drive mechanisms between the loading slide and the loading drive motor and between the washing slide and the washing drive motor are not limited to transmission via the drive belt, screw drive assembly, or the like described above.
The liquid pumping mechanism 400 is connected to the sample addition needle 210, the wash basin 110, the filling needle 310, and the pipetting needle 320, respectively.
The control mechanism 500 is connected to the sample loading drive device 220, the sample washing drive device, and the liquid pumping mechanism 400 to control the operations of the sample loading drive device 220, the sample washing drive device, and the liquid pumping mechanism 400, respectively.
Specifically, in the present embodiment, the control mechanism 500 is used to control the loading drive device 220 to move the loading needle 210 to the reagent zone 102, and to control the liquid pumping mechanism 400 to pump the reagent from the reagent zone 102 via the loading needle 210. The control mechanism 500 is also used to control the loading drive 220 to move the loading needle 210 to the incubation area 106 and to control the pumping mechanism 400 to pump the reagent from the loading needle 210 into the reaction vessel in the incubation well 120. The control mechanism 500 is also used to control the loading drive device 220 to move the loading needle 210 into the wash basin 110, and to control the pumping mechanism 400 to pump the wash liquid from the wash liquid in the wash liquid zone 108 to wash the inner and outer walls of the loading needle 210, respectively. The control mechanism 500 is also used to control the loading drive device 220 to reset the loading needle 210. The control mechanism 500 is also used for controlling the liquid pumping mechanism 400 to pump out the cleaning waste liquid in the needle washing tank 110. The control mechanism 500 is also used to control the sample injection drive to move the injection needle 310 to the incubation zone 106 and to control the pump mechanism 400 to pump the wash liquid from the wash liquid in the wash liquid zone 108 and out through the injection needle 310 to the reaction vessel. The control mechanism 500 is also used for controlling the sample washing driving device to move the pipetting needle 320 to the incubation area 106 and controlling the pipetting mechanism 400 to pump out the washing waste liquid in the reaction vessel via the pipetting needle 320. The control mechanism 500 is also used for controlling the sample washing driving device to respectively move the filling needle 310 and the pipetting needle 320 into the washing needle basin 110, and controlling the liquid pumping mechanism 400 to pump the washing liquid from the washing liquid in the washing liquid area 108 so as to respectively wash the filling needle 310 and the pipetting needle 320. The control mechanism 500 is also used for controlling the sample washing driving device to drive the liquid injection needle 310 and the liquid suction needle 320 to reset.
In one embodiment, the reagent zone 102 has a reagent rack 130. A reagent bottle may be placed in the reagent rack 130. Preferably, there are a plurality of reagent racks 130 for placing reagent bottles of different sizes.
In one embodiment, the bottom of the reagent rack 130 is provided with a first heating device (not shown). The first heating means is connected to the control mechanism 500 for heating the reagent bottles 30 placed in the reagent rack 130 to control the temperature of the reagent to be added.
The top of the incubation slot 120 is provided with a lid 122. As shown in fig. 5, 6 and 7, in one embodiment, the lid 122 is slidably disposed on the lid slide rail 124 and correspondingly also includes a lid driver 126. The lid driving device 126 is used to drive the lid 122 to move along the lid sliding rail 124 to close or open the incubation slot 120. More specifically, in this embodiment, the lid driver 126 is drivingly connected to the lid 122 by a belt drive assembly 128. Further, the back of the cover 122 is adhered with a heat insulating material layer, such as EVA layer, to reduce heat dissipation, and the incubation slot 120 is closed to reduce evaporation of liquid during incubation.
In one embodiment, the bottom of the incubation slot 120 is provided with a second heating means (not shown). The second heating device is connected with the control mechanism 500 to maintain or control the temperature in the incubation groove 120 in real time, so as to ensure the stable incubation process. Further, a temperature sensor is provided in the incubation groove 120. The temperature sensor is electrically connected to the control mechanism 500 to monitor the temperature in the incubation slot 120 in real time.
In addition, in one embodiment, a jacking mechanism is further provided at the bottom of the incubation slot 120. The jacking mechanism is connected with the control mechanism 500 to jack up one side of the reaction vessel 20 in the incubation groove by the control mechanism 500 to incline so as to facilitate the liquid suction of the liquid suction needle 320. Referring to fig. 6, 7 and 8, in one embodiment, there are two incubation slots 120, and one reaction vessel 20 may be placed in each incubation slot 120. Correspondingly, the jacking mechanism comprises two jacking columns 121 arranged at the bottom of each incubation groove 120. Four jacking columns 121 are arranged in the two incubation grooves 120. Each top column 121 is independently controlled to be telescopic in the vertical direction, for example, the top column 121 may be an output shaft of a telescopic cylinder, or may be driven to ascend or descend by a driving motor in a forward and reverse rotation manner. The two top posts 121 in each incubation groove 120 respectively correspond to two sides of the reaction container 20, and can be respectively telescopic, so that the swinging and tilting of the reaction container 20 can be realized, and the incubation liquid can be fully mixed.
Referring to fig. 9 and 10, in one embodiment, the wash zone 108 is provided with a plurality of wash bottles 40. The plurality of wash liquid bottles 40 are connected to the liquid pumping mechanism 400 by pipes, respectively. An independent control valve 460 is arranged on the corresponding pipeline of each lotion bottle 40, and the control valve 460 is preferably connected with a control mechanism so that the control mechanism can automatically and independently control the on or off of each pipeline to facilitate the pumping of different cleaning liquids.
In one embodiment, the reaction vessel 20 is preferably a multi-well plate, and accordingly, the size and shape of the incubation well 120 corresponds to the multi-well plate. In response, the injection needles 310 and the pipetting needles 320 are arranged in rows, respectively, and the plurality of injection needles 310 in rows are spaced apart from the plurality of pipetting needles 320 in rows. For the injection needles 310 and the aspiration needles 320 arranged in rows, they are arranged in parallel rows, such as along the X-axis. The injection needles 310 and the pipetting needles 320 are one high and one low and one thick and one thin, specifically, the injection needles 310 of the first row are thin and short, and the pipetting needles 320 of the second row are thick and long. The filling needle 310 and the suction needle 320 are arranged in a row, and the cleaning liquid can be efficiently and conveniently added and sucked out.
In a specific embodiment, as shown in fig. 11 and 12, the reaction vessel 20 is a 24-well plate, and accordingly, the injection needles 310 and the pipetting needles 320 are respectively arranged on the needle holder 340 in rows, and 6 injection needles 310 or 6 pipetting needles 320,6 injection needles 310 or 6 pipetting needles 320 are arranged at intervals in each row. The needle seat 340 is provided with two pipeline interfaces 342, and the two pipeline interfaces 342 are respectively communicated with the liquid injection needle 310 and the liquid suction needle 320 through independent flow passages inside the needle seat 340.
In one embodiment, there are two needle wash zones 104, each needle wash zone 104 being provided with a needle wash basin 110. One of the wash wells 110 is smaller in size and positioned adjacent to the reagent zone 102 for washing the sample application needle 210, and the other wash well 110 is larger in size and positioned behind the incubation zone 106 for washing the pipetting needle 310 and the pipetting needle 320. The wash drive means may drive the injection needle 310 and the pipetting needle 320 between the incubation area 106 and the wash basin 110 disposed behind the incubation area 106, as in the embodiment of fig. 5, between the incubation area 106 and the respective wash basin 110 by a Y-axis wash motor 335. As shown in fig. 10, in one embodiment, the pumping mechanism 400 includes a plunger pump 450 and a first diaphragm pump 410. The plunger pump 450 and the first diaphragm pump 410 are connected in series with the sample application needle 210. Plunger pump 450 is used to control the pumping of reagent from reagent zone 102 by sample addition needle 210 and to control the pumping of reagent by sample addition needle 210 into reaction vessel 20. The first diaphragm pump 410 is used to pump cleaning fluid from the wash zone 108 into the loading needle 210 and out into the wash well 110 to clean the interior and exterior walls of the loading needle 210.
Further, in one embodiment, the pumping mechanism 400 further includes a second diaphragm pump 420. The second diaphragm pump 420 is connected in series with the needle wash basin 110. The second diaphragm pump 420 is used to pump out the waste liquid of the needle wash basin 110.
Still further, in one embodiment, the pumping mechanism 400 further includes a third diaphragm pump 430 and a fourth diaphragm pump 440. A third diaphragm pump 430 is connected in series with the fill needle 310 to pump cleaning fluid from the cleaning fluid zone 108 and out through the fill needle 310. A fourth diaphragm pump 440 is connected to the pipetting needle 320 to pump the washing waste liquid from the reaction vessel through the pipetting needle 320 and out.
In one embodiment, the nucleic acid hybridization apparatus 10 further comprises a waste bottle interface 600 provided on the machine 100. The waste liquid bottle interface 600 is connected to the second diaphragm pump 420 and the fourth diaphragm pump 440 to discharge the washing waste liquid.
In addition, as shown in fig. 9, in a specific embodiment, the machine 100 is provided with a cover 140, and the cover 140 covers the elements inside. The cover 140 is provided with a first protective door 142 and a second protective door 144. After the first guard door 142 is opened, reagents may be placed in the reagent zone 102, the reaction vessel 20 placed in the incubation zone 106, etc.; after the second guard door 144 is opened, the wash bottle 40 can be placed therein. Through setting up the cover body 140, can effectively prevent external factor to incubating the influence of experimental process, guarantee the reliability of experimental operation.
In a preferred embodiment of the nucleic acid hybridization apparatus, the workflow in performing in situ hybridization is as follows:
1. and (3) adding a reagent: the sample loading arm group drives the sample loading needle 210 to move, the sample loading needle 210 is moved into the reagent bottle 30 on the reagent rack 130, and the plunger pump 450 acts to suck quantitative reagents. The loading arm set then moves the loading needle 210 onto the sample in the incubation slot 120, and the plunger pump 450 moves to push the reagent out of the loading needle 210 and onto the sample.
2. Sample addition needle 210 cleans: the sample loading arm group drives the sample loading needle 210 to be inserted into the needle washing pool 110, a first diaphragm pump 410 connected with the sample loading needle 210 is started, cleaning liquid is pumped to clean the inner wall of the sample loading needle 210, the cleaning liquid is simultaneously discharged into the needle washing pool 110 to clean the outer wall of the sample loading needle 210, cleaning waste liquid flows into the needle washing pool 110, a second diaphragm pump 420 connected with the needle washing pool 110 is started again, and cleaning waste liquid is pumped into a waste liquid barrel through a waste liquid bottle interface 600 by using the second diaphragm pump 420.
3. Incubation: when the reaction vessel 20 is incubated in the incubation well 120, the lid 122 will automatically slide out to cover the entire incubation well. The procedure of incubation at room temperature does not need temperature adjustment, and the procedure of incubation at heating is performed after the bottom of the incubation groove 120 is preheated to a designated temperature under the control of the control mechanism 500.
4. And (3) cleaning solution addition: the cleaning solution is added by means of a sample washing mechanism 300, taking a nucleic acid hybridization instrument with a 24-hole plate as an example for the reaction vessel 20, six liquid injection needles 310 are arranged in parallel in the sample washing mechanism 300, the sample washing arm group drives the liquid injection needles 310 to move to the position right above the 24-hole plate, the communication with the corresponding cleaning solution bottle 40 is realized through a control valve 460 gate switch, a third diaphragm pump 430 is started again, the six liquid injection needles 310 absorb quantitative cleaning solution into the 24-hole plate under the drive of the third diaphragm pump 430, and finally the sample injection needles 310 are subjected to sample washing and needle arrangement recovery under the drive of the sample washing arm group belt.
5. And (3) cleaning solution removal: the washing solution is removed by sucking the washing solution by the sucking needle 320, and the sample washing mechanism 300 is provided with six sucking needles 320 side by side, as in the case of the nucleic acid hybridization instrument in which the reaction vessel 20 is a 24-well plate. The jack-up of incubation groove 120 bottom is with 24 orifice plates jack-up slope, washes appearance arm group and drives the liquid suction needle 320 and insert 24 orifice plates liquid level under, starts fourth diaphragm pump 440, six liquid suction needles 320 are driven by fourth diaphragm pump 440 and are absorbed 24 orifice plates the washing liquid in the orifice plate, finally wash appearance arm group drive liquid suction needle 320 and resume the normal position.
6. Hybridization: the nucleic acid hybridization apparatus 10 can be properly adjusted in terms of the operation time and the reagent amount of each step according to the hybridization program set in the control mechanism 500 (e.g., PLC controller, etc.), and can achieve effective accuracy within a certain range. Various reagents were added to the 24-well plate sample wells by pipetting mechanism 200 and pipetting mechanism 400. The sample washing mechanism 300 and the liquid pumping mechanism 400 are used for realizing the liquid adding and removing operation of the cleaning liquid; preheating and hybridization can be achieved by further controlling the temperature of the incubation well 120 by the control mechanism 500. And taking out the sample after hybridization, staining, and observing the detection result by a fluorescence microscope.
The nucleic acid hybridization instrument 10 can realize full-automatic experimental operation in the whole process by setting a hybridization program, is different from manual operation in the process of complicated hybridization steps, avoids consuming a great deal of manpower and time, eliminates errors caused by human factors, and saves a great deal of experimental time. The independent reagent adding and cleaning regulation device and the cleaning liquid adding and sucking regulation device are adopted, and the control mechanism is used for controlling the corresponding driving device to accurately move, so that the rapid and convenient operation and the accurate liquid adding and sucking operation are realized; the constant-temperature heating is controlled by the control mechanism, the heating is direct and rapid, the incubation temperature and the effective retention of the reaction reagent are ensured by the sealing of the box cover, the experimental accuracy is effectively improved, and the experimental error is reduced.
The technical features of the above-described embodiments may be arbitrarily combined, and all possible combinations of the technical features in the above-described embodiments are not described for brevity of description, however, as long as there is no contradiction between the combinations of the technical features, they should be considered as the scope of the description.
The above examples illustrate only a few embodiments of the invention, which are described in detail and are not to be construed as limiting the scope of the invention. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the invention, which are all within the scope of the invention. Accordingly, the scope of protection of the present invention is to be determined by the appended claims.
Claims (2)
1. A nucleic acid hybridization apparatus, comprising:
the machine comprises a reagent zone, a needle washing zone, an incubation zone and a washing liquid zone, wherein the reagent zone is used for placing a reagent, the needle washing zone is provided with a needle washing pool, the incubation zone is provided with an incubation groove used for placing a reaction container, and the washing liquid zone is used for placing a washing liquid;
the sample adding driving device is arranged on the machine table and connected with the sample adding needle to drive the sample adding needle to move among the reagent zone, the needle washing zone and the incubation zone;
the sample washing mechanism comprises a liquid injection needle, a liquid suction needle and a sample washing driving device, wherein the sample washing driving device is arranged on the machine table and is respectively connected with the liquid injection needle and the liquid suction needle to be used for driving the liquid injection needle and the liquid suction needle to move between the needle washing area and the incubation area;
the liquid pumping mechanism is respectively connected with the sample adding needle, the needle washing pool, the liquid injection needle and the liquid suction needle; and
the control mechanism is connected with the sample adding driving device, the sample washing driving device and the liquid pumping mechanism;
the control mechanism is used for controlling the sample adding driving device to move the sample adding needle to the reagent area and controlling the liquid pumping mechanism to pump reagent from the reagent area through the sample adding needle; the control mechanism is also used for controlling the sample adding driving device to move the sample adding needle to the incubation area and controlling the liquid pumping mechanism to pump out the reagent in the sample adding needle to the reaction container in the incubation groove; the control mechanism is also used for controlling the sample adding driving device to move the sample adding needle into the needle washing pool and controlling the liquid pumping mechanism to pump the cleaning liquid from the cleaning liquid in the cleaning liquid area so as to clean the inner wall and the outer wall of the sample adding needle respectively; the control mechanism is also used for controlling the sample adding driving device to reset the sample adding needle; the control mechanism is also used for controlling the liquid pumping mechanism to pump out the cleaning waste liquid in the needle washing pool; the control mechanism is also used for controlling the sample washing driving device to move the liquid injection needle to the incubation area and controlling the liquid pumping mechanism to pump the cleaning liquid from the cleaning liquid in the cleaning liquid area and pump the cleaning liquid to the reaction container through the liquid injection needle; the control mechanism is also used for controlling the sample washing driving device to move the liquid sucking needle to the incubation area and controlling the liquid pumping mechanism to pump the washing waste liquid in the reaction container out through the liquid sucking needle; the control mechanism is also used for controlling the sample washing driving device to respectively move the liquid injection needle and the liquid suction needle into the needle washing pool, and controlling the liquid pumping mechanism to pump cleaning liquid from the cleaning liquid in the washing liquid area so as to respectively clean the liquid injection needle and the liquid suction needle; the control mechanism is also used for controlling the sample washing driving device to drive the liquid injection needle and the liquid suction needle to reset;
the reagent zone is provided with a reagent rack; the bottom of the reagent rack is provided with a first heating device which is connected with the control mechanism;
the bottom of the incubation groove is provided with a second heating device, and the second heating device is connected with the control mechanism;
the bottom of the incubation groove is provided with a jacking mechanism, and the jacking mechanism is connected with the control mechanism so as to control one side of the reaction container in the incubation groove to be jacked by the control mechanism to form an inclination;
a temperature sensor is arranged in the incubation groove and is electrically connected with the control mechanism;
the top of the incubation groove is provided with a box cover;
the box cover driving device is connected with the box cover and used for opening or closing the incubation groove by driving the box cover to move;
the washing liquid area is provided with a plurality of washing liquid bottles which are respectively connected with the liquid pumping mechanism through pipelines; an independent control valve is arranged on the pipeline corresponding to each washing liquid bottle, and the control valve is connected with the control mechanism so as to be controlled to be opened or closed by the control mechanism;
the liquid pumping mechanism comprises a plunger pump and a first diaphragm pump, the plunger pump and the first diaphragm pump are connected in series with the sample adding needle, the plunger pump is used for controlling the sample adding needle to pump reagent from the reagent zone and controlling the sample adding needle to pump the pumped reagent into the reaction container, and the first diaphragm pump is used for pumping cleaning liquid from the cleaning liquid zone into the sample adding needle and pumping the cleaning liquid into the needle washing pool so as to clean the inner wall and the outer wall of the sample adding needle;
the liquid pumping mechanism further comprises a second diaphragm pump, the second diaphragm pump is connected with the needle washing pool in series, and the second diaphragm pump is used for pumping out waste liquid of the needle washing pool;
the liquid pumping mechanism further comprises a third diaphragm pump and a fourth diaphragm pump, the third diaphragm pump is connected with the liquid injection needle in series to pump cleaning liquid from the cleaning liquid area and pump the cleaning liquid through the liquid injection needle, and the fourth diaphragm pump is connected with the liquid suction needle in series to pump cleaning waste liquid from the reaction container through the liquid suction needle and pump the cleaning waste liquid out;
the device also comprises a waste liquid bottle interface arranged on the machine table, and the waste liquid bottle interface is connected with the second diaphragm pump and the fourth diaphragm pump.
2. The nucleic acid hybridization instrument according to claim 1, wherein the injection needle and the pipetting needle are arranged in rows, respectively, and a plurality of the injection needles in a row are arranged at intervals from a plurality of the pipetting needles in a row.
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