CN112051122A

CN112051122A - Sample applicator

Info

Publication number: CN112051122A
Application number: CN202010967575.6A
Authority: CN
Inventors: 汪云飞; 潘能科
Original assignee: Chongqing Topu Bioengineering Co Ltd
Current assignee: Chongqing Topu Bioengineering Co Ltd
Priority date: 2020-09-15
Filing date: 2020-09-15
Publication date: 2020-12-08

Abstract

The invention discloses a sample applicator, which comprises a frame, a sample application device, a drying and storing device, a reagent storing device and a liquid path device, wherein the sample application device, the drying and storing device, the reagent storing device and the liquid path device are all arranged on the frame; the sample application device comprises a target plate platform for mounting a target plate, a sample application needle for sample application and a driving device for driving the target plate platform or the sample application needle; the drying and storing device is arranged on the motion track of the target plate platform to dry the target plate; the reagent storage device is arranged on the motion track of the sample application needle. Through the triaxial interlock motion that sets up XYZ axle motion subassembly, realized the accurate sample application of full automatization of injector to the target plate, very big improvement work efficiency. Through setting up the disinfection storehouse, utilize the alcohol spraying to disinfect to the target plate, play the effect of inactivation microorganism, furthest's reduction pollutes, has improved the rate of accuracy that detects.

Description

Sample applicator

Technical Field

The invention relates to sample pretreatment, in particular to a sample applicator.

Background

The operating principle of MALDI-TOF-MS is that matrix-assisted laser desorption ionization is utilized to analyze components on the surface of a sample to be detected into ions, and the ions with different mass-to-charge ratios (m/z) are separated through a flight time mass analyzer according to the length of the flight time to form mass spectrum peaks (mass spectrograms) with different intensities. In the field of microorganism identification, a bacterial or fungal mass spectrum database is established by using a standard strain and a strain with clear source typing, and the mass spectrum of an unknown strain is compared with the database by using MassLynx software (or other related software), so that the species of a microorganism to be detected can be determined. Compared with other microorganism detection technologies, the MALDI-TOF-MS technology has obvious high efficiency and accuracy, and the cost of each experiment is very low, so that the MALDI-TOF-MS technology becomes a popular method for microorganism identification.

MALDI-TOF-MS mass spectrometry is mainly composed of two parts: matrix-assisted laser desorption ionization ion source (MALDI) and time-of-flight mass analyzer (TOF). The matrix-assisted laser desorption ion source is used for desorbing a solid sample, the sample is manufactured under normal pressure to form a solid, the solid enters a vacuum system through a sample injection mechanism, and the solid is accurately positioned by a positioning mechanism so that the sample is desorbed in a specific area. During the analysis process, the laser provides high energy to the sample in a very short time, and the sample is heated very quickly, so that the matrix molecules in the ion source can effectively absorb the energy of the laser and indirectly transfer the energy to the sample molecules, the sample molecules are ionized, and then the ions enter a detector under the action of high pressure for detection.

The matrix-assisted laser desorption (MALDI) ion source sampling apparatus is critical to the overall apparatus. The sample to be measured (mostly biological sample) can not directly follow the ordinary pressure environment in extremely short time internal conversion become the high vacuum environment, consequently sampling device need guarantee that the sample gradually enters into the high vacuum environment from the ordinary pressure environment, lets the testing sample have an adaptation process, just so can not lead to the fact big influence to the sample analysis. Usually, the sample is first fixed on the target spot of the target plate and then sent into a matrix-assisted laser desorption ionization time-of-flight mass spectrometer (MALDI-TOF) for analysis and identification. However, the current operation of fixing the sample on the target plate is generally performed manually, which is not only inefficient, but also has low spotting accuracy and even affects the final mass spectrometer analysis result.

The prior art has disclosed the structure of a fully automatic sample applicator, which has a longitudinal (Y axis) and vertical (Z axis) and transverse (X axis) three-axis motion assembly, and realizes sample application of a target plate by the linkage of the three axes XYZ. However, the sample application process of the sample application instrument is completely exposed in the air, and microorganisms in the environment are easily introduced, so that the detection result is influenced.

Disclosure of Invention

Aiming at the defects of the prior art, the invention provides a sample application instrument, which solves the problem that the subsequent detection result has errors due to microbial pollution in the sample application process.

In order to achieve the purpose, the invention is realized by the following technical scheme:

a spotting instrument comprises a frame and a spotting device, and is characterized in that: the sample application device, the drying and storing device and the reagent storing device are all arranged on the frame;

the sample application device comprises a target plate platform for mounting a target plate, a sample application needle for sample application and a driving device for driving the target plate platform or the sample application needle to move;

the drying and storing device is arranged on the motion trail of the target plate platform and is used for drying the target plate;

the reagent storage device is arranged on the motion track of the sample application needle.

Preferably, the rack comprises an outer shell and a cabin door rotationally connected with the outer shell, the outer shell encloses an inner space of the rack, the cabin door and the outer shell enclose an operation space, the inactivation device and the reagent storage device are positioned in the operation space, and the dry preservation device is positioned in the inner space of the rack;

the upper surface of the shell is downwards sunken to form a mounting part for mounting a container;

the cabin door is provided with an observation window.

Preferably, the driving device comprises an X-direction moving assembly, a Y-direction moving assembly and a Z-direction moving assembly;

the X-direction movement assembly comprises an X-direction motor, an X-direction slide rail, a screw rod and an X-direction sliding block, the screw rod is connected with the X-direction motor in a matched mode, the X-direction sliding block is installed on the X-direction slide rail and connected with the screw rod in a matched mode, and the X-direction motor drives the X-direction sliding block to move along the X direction;

the Z-direction movement assembly comprises a Z-direction motor and a Z-direction slide rail which are arranged on the X-direction slide block, a screw rod which is connected with the Z-direction motor in a matched mode, and a Z-direction slide block which is arranged on the Z-direction slide rail and connected with the screw rod in a matched mode, and the Z-direction motor drives the Z-direction slide block to move along a Z axis;

the Y-direction movement assembly comprises a Y-direction motor, a Y-direction slide rail, a screw rod and a Y-direction sliding block, the screw rod is connected with the Y-direction motor in a matched mode, the Y-direction sliding block is installed on the Y-direction slide rail and connected with the screw rod in a matched mode, and the Y-direction motor drives the Y-direction sliding block to move along the Y axis;

the sample application needle is arranged on the Z-direction sliding block, and the target plate platform is connected with the Y-direction sliding block.

Preferably, the spotting device further comprises a positioning device, the positioning device comprises a positioning switch and a positioning plate, the positioning plate is provided with a plurality of protrusions and a plurality of recesses, and the positions of the positioning switch and the positioning plate are matched.

Preferably, the protruding portions and the recessed portions on the positioning plate are uniformly distributed at intervals, and the positioning plate is serrated.

Preferably, the positioning plates are distributed along the Y axis, and the positioning switches are arranged on the Y-direction sliding blocks; or the positioning plates are distributed along the X direction, and the positioning switches are arranged on the X-direction sliding blocks.

Preferably, the spotting device further comprises a limiting device, the limiting device comprises a limiting part and a limiting switch, the limiting part is fixed on the X-direction sliding block and/or the Y-direction sliding block and/or the Z-direction sliding block, and the limiting switch is fixed on the X-axis and/or the Y-axis and/or the Z-axis at a position matched with the limiting part.

Preferably, the inactivation device comprises a sealing cover, an atomizer and a pump, wherein the sealing cover is used for covering the target plate platform, the sealing cover is arranged on the cabin door, the atomizer and the pump are both arranged on the rack, the sealing cover and the target plate platform are matched to form an inactivation space, and the wall of the inactivation space is provided with an air inlet and an air outlet; the atomizer is equipped with the inactivation liquid in to can the atomizing inactivation liquid, the pump is used for providing power drive inactivation fog and discharges through the gas outlet after the air inlet moves to the inactivation space.

Preferably, the inactivation space formed by the sealing cover and the target plate platform is provided with a slit.

Preferably, the target plate inactivation device further comprises a tail gas treatment bottle, wherein the tail gas treatment bottle is connected with the gas outlet and is used for collecting the inactivation mist discharged through the inactivation space; the pump is connected with the tail gas treatment bottle and is used for providing power to drive the inactivation mist in the inactivation space to be discharged into the tail gas treatment bottle through the gas outlet;

the device comprises two pumps, one of the two pumps is connected with the atomizer and used for providing power to drive the inactivation mist generated by the atomizer to move to the inactivation space through the air inlet, and the other pump is connected with the tail gas treatment bottle and used for providing power to drive the inactivation mist in the inactivation space to be discharged into the tail gas treatment bottle through the air outlet;

the side wall of the sealing cover is provided with a notch, the air inlet and the air outlet are arranged on the shell, and the positions of the air inlet and the air outlet correspond to the positions of the notch.

Preferably, dry save set includes support, closing plate, vacuum pump and cam mechanism, target plate platform and support set up on the base, closing plate, vacuum pump and cam mechanism set up on the support, be provided with the opening that corresponds with cam mechanism position on the support, cam mechanism acts on the closing plate, and the closing plate forms confined space with the target plate platform, the vacuum pump is used for extracting the interior air of confined space.

Preferably, the cam structure comprises a cam, a drying motor and a cam mounting frame, the cam and the drying motor are arranged on the cam mounting frame, and the cam is connected with a rotating shaft of the drying motor;

the cam is provided with a remote angle of repose of 20-40 degrees;

a bearing is arranged at the maximum stroke position of the cam;

the cam structure further comprises a sensor, the sensor comprises a trigger piece and an inductive switch, the trigger piece is arranged at the bottom of the cam, and the inductive switch is arranged on the cam mounting frame and is matched with the trigger piece;

the sealing plate is arranged on the bracket through a threaded rod, a spring is sleeved on the threaded rod, and the elastic force of the spring presses the sealing plate on the bracket; the springs and the threaded rods are respectively 4 and are respectively arranged at four corners of the sealing plate;

the sealing plate is provided with a trapezoidal groove, and an O ring is arranged in the trapezoidal groove; or a trapezoidal groove is formed in the target groove, and an O ring is arranged in the trapezoidal groove;

the sealing plate is provided with a threaded hole, the threaded hole is provided with a quick-screwing straight-inserting connector, one electromagnetic valve is connected with the air pumping hole of the vacuum pump, the other electromagnetic valve is connected with a three-way connector, and the other two holes of the three-way connector are respectively connected with the air inlet of the vacuum pump and the quick-screwing straight-inserting connector.

Preferably, the reagent storage device comprises a reagent storage structure mounted on the rack, a reagent sealing motor, a connecting structure connected with the reagent sealing motor in a matching manner, and a cover connected with the connecting structure, wherein one or more reagent holes for storing reagents are formed in the reagent storage structure.

Preferably, the direction of movement of the lid is at an angle to the upper surface of the reagent storage device;

the cover is made of aluminum alloy materials, and a sealing piece is fixed on the surface of the cover facing the reagent storage structure;

the reagent sealing motor is a rotating motor, and the axis of the rotating shaft of the reagent sealing motor is in the horizontal direction;

still include support and sensor, the sensor is installed on the support, the support is fixed in the frame, connection structure is last to have the trigger piece, the sensor is with the position looks adaptation of trigger piece.

Preferably, the needle washing device comprises a washing tank, a liquid inlet peristaltic pump and a liquid discharge peristaltic pump, the needle washing tank is provided with a side wall and a bottom wall, a liquid inlet is formed in the side wall, a liquid discharge port is formed in the bottom wall, the liquid inlet peristaltic pump and the liquid discharge peristaltic pump are both arranged on the shell, one end of the liquid inlet peristaltic pump is located in the operation space, and the other end of the liquid inlet peristaltic pump is located in the machine space; one end of the liquid discharge peristaltic pump is positioned in the operation space, and the other end of the liquid discharge peristaltic pump is positioned in the machine space; the liquid inlet peristaltic pump is connected with the liquid inlet, and the liquid discharge peristaltic pump is connected with the liquid discharge port.

Preferably, the length of the liquid inlet is more than 1mm, the axis extension line of the liquid inlet deviates from the central axis of the needle washing groove, the cross section of the inner wall of the side wall is circular or elliptical, and the bottom wall is conical.

The invention provides a sample applicator with self-disinfecting and sterilizing capability, which has the following beneficial effects:

(1) the sample application needle is provided with the disinfection device, the target plate can be disinfected before sample application, the situation that the target plate is provided with organisms in the environment can be avoided, the situation that the microorganisms in the environment influence a test result is avoided, and meanwhile, the sample application needle can automatically clean the needle washing device along the X-axis motion track after sample application of the target plate is completed, so that the pollution of a sample is avoided to the maximum extent, and the accuracy of a detection result is ensured.

(2) According to the invention, through the three-axis synergistic effect of the X-axis motion assembly, the Y-axis motion assembly and the Z-axis motion assembly, the full-automatic accurate substrate covering of the target plate is realized, and meanwhile, the drying and storing device is arranged on the Y-axis motion track, so that the target plate can be dried for multiple times in a vacuum drying mode, and the working efficiency of sample application is greatly improved.

(3) The sample application instrument is provided with an organic inner space and an operation space, only the target plate platform, the sample application needle and the reagent storage device which are frequently required to be operated are arranged in the operation space, and the operation space is relatively closed, so that the sample application instrument is better protected, dust and abrasion are avoided, meanwhile, the target plate is always in a relatively closed environment after being inactivated, and the risk that the target plate is polluted by microorganisms in the environment again is reduced.

(4) The container is arranged at the top of the shell, so that the condition in the container can be observed conveniently, the container can be filled with liquid, replaced or cleaned conveniently, and the liquid can flow under the action of gravity. The installation part sunken downwards on the shell can provide the installation position of the container, and can keep the container stable to prevent the container from falling.

Drawings

FIG. 1 is a schematic view of a spotting instrument of the present invention;

FIG. 2 is a schematic view of the spotting device of the present invention;

FIG. 3 is a rear view of the hidden enclosure and door of FIG. 1;

FIG. 4 is a schematic view of the structure of the sealed space of the present invention;

FIG. 5 is a schematic view of the dry storage apparatus of the present invention;

FIG. 6 is a schematic view of the connection and mode of operation of the vacuum pump of the present invention;

FIG. 7 is a schematic view of a reagent storage device according to the present invention;

FIG. 8 is a schematic view of the structure of the needle washing device of the present invention.

Reference numerals in the drawings of the specification include:

the device comprises a shell 11, a cabin door 12, an observation window 13 and a mounting part 14;

the device comprises a target plate platform 21, a sample application needle 22, an X-direction motor 231, an X-direction slide rail 232, an X-direction slide block 233, a Y-direction motor 241, a Y-direction slide rail 242, a Y-direction slide block 243, a Z-direction motor 251, a Z-direction slide rail 252, a Z-direction slide block 253, a positioning switch 261, a positioning plate 262, a limit switch 271 and a limit piece 272;

the device comprises a sealing cover 31, a notch 311, an air inlet 312, an air outlet 313, an atomizer 32, a tail gas treatment bottle 33, a first diaphragm pump 34 and a second diaphragm pump 35;

the device comprises a support 41, an opening 411, a sealing plate 42, a spring 421, a threaded rod 422, a quick-screwing straight plug 423, a vacuum pump 43, a first switch 44, a second switch 45, a cam 461, a drying motor 462, a cam mounting rack 463, a first induction switch 464 and a first trigger piece 465;

the reagent storage structure 51, the cover 54, the connecting structure 53, the second inductive switch 531 and the second trigger piece 532;

a needle washing groove 61, a liquid inlet 611 and a liquid outlet 612.

Detailed Description

The present invention will be further described with reference to the following drawings and examples, which are illustrative only and not intended to be limiting, and the scope of the present invention is not limited thereby.

A sample applicator comprises a frame, a sample application device, an inactivation device, a drying and storing device, a reagent storing device and a needle washing device.

As shown in fig. 1, the rack includes an outer shell 11 and a container mounting base plate 14, a cabin door 12 is movably connected to the outer shell 11, in this embodiment, the cabin door 12 is rotatably connected to the outer shell 11, the outer shell 11 encloses an inner space, the cabin door 12 and the outer shell 11 enclose an operating space, an observation window 13 is provided on the cabin door 12 for observing a state in the operating space, and an upper surface of the outer shell 11 is recessed downward to form a mounting portion 14 for mounting a container.

The inactivation device is arranged on one side of the target plate platform and used for inactivating the target plate; the drying and storing device is arranged on the motion trail of the target plate platform and used for drying the target plate; the reagent storage device and the needle washing device are arranged on the motion track of the sample application needle 22; the needle washing device, the reagent storage device and the inactivation device are all positioned in the operation space; the drying and storing device and the X-direction moving assembly are positioned in the machine space; the print needle 22 is located and only moves within the process space; the target plate platform 21 moves in the operation space and the machine space under the drive of the Y-direction moving assembly.

The horizontal direction of the horizontal plane of the bottom plate is taken as an X axis, the longitudinal direction is taken as a Y axis, and the axis vertical to the bottom plate is taken as a Z axis.

The invention has sample application device, inactivation device, drying and storing device, reagent storing device and needle washing device, and can inactivate, sample application, dry and store the target plate, and realize automatic sample application.

The invention has the built-in space and the operation space, so that the built-in space can ensure that most parts are in a better sealed environment and are better protected, the operation space can facilitate the operations of taking and placing the target plate, adding reagents, spotting and the like, and the working environment can not be exposed in the external environment. Meanwhile, the arrangement of the operation space also creates conditions for the arrangement of the inactivation space, and the fog overflowing from the inactivation space can not be directly emitted to the outside to cause pollution.

The container is arranged at the top of the shell, so that the condition in the container can be observed conveniently, the container can be filled with liquid, replaced or cleaned conveniently, and the liquid can flow under the action of gravity. The installation part sunken downwards on the shell can provide the installation position of the container, and can keep the container stable to prevent the container from falling.

As shown in fig. 2, the spotting device comprises: a target plate platform 21 for mounting the target plate, a sample application needle 22 for sample application, a driving device for driving the target plate platform or the sample application needle, a syringe pump, a positioning device for positioning the movement and a limiting device.

The driving device comprises an X-direction movement assembly, a Y-direction movement assembly and a Z-direction movement assembly;

the X-direction movement assembly comprises an X-direction motor 231, an X-direction slide rail 232, a screw rod in fit connection with the X-direction motor, and an X-direction slider 233 which is arranged on the X-direction slide rail 232 and in fit connection with the screw rod, wherein the X-direction motor 231 drives the X-direction slider 233 to move along the X direction;

the Z-direction movement assembly comprises a Z-direction motor 251 and a Z-direction slide rail 252 which are arranged on the X-direction slide block, a screw rod which is matched and connected with the Z-direction motor 251, and a Z-direction slide block 253 which is arranged on the Z-direction slide rail 252 and is matched and connected with the screw rod, wherein the Z-direction motor 251 drives the Z-axis slide block 253 to move along the Z axis;

the Y-direction movement assembly comprises a Y-direction motor 241, a Y-direction slide rail 242, a screw rod in fit connection with the Y-direction motor 241, and a Y-direction slider 243 which is arranged on the Y-direction slide rail 242 and in fit connection with the screw rod, wherein the Y-direction motor 241 drives the Y-direction slider 243 to move along the Y axis;

the sample application needle 22 is mounted on the Z-direction slide 253, the injection pump drives the sample application needle 22 to sample and apply samples, and the target plate platform 21 is connected with the Y-direction slide 243.

The positioning device comprises positioning switches 261 and positioning plates 262, the number is two sets, a plurality of protruding portions and a plurality of corresponding recessed portions which are evenly distributed at intervals are arranged on the positioning plates 262, a saw-toothed structure is formed, the two positioning plates 262 are respectively arranged on an X axis and a Y axis, the two positioning switches 261 are respectively arranged on the X direction sliding block 233 and the Y direction sliding block 243, and the positions of the positioning switches 261 are matched with the positioning plates 262 one by one (the matching mentioned here refers to the fact that the protruding portions can trigger the positioning switches 261 or the recessed portions can trigger the positioning switches 272). In this embodiment, the positioning switches 261 are all photoelectric switches.

The accurate positioning of the sample application needle in the X direction and the target plate motion platform in the Y direction is controlled by the matching of the positioning switch 261 and the positioning plate 262. Since the positioning plate 262 has a plurality of protrusions and a plurality of depressions, a plurality of positions can be positioned, thereby positioning all points on the target plate.

Under positioner's supplementary effect, can carry out accurate application of sample with the mode of liquid contact (application of sample needle contactless target board), and then furthest has avoided cross contamination, has reduced the needle washing number of times simultaneously, has improved work efficiency, and after the solution application of sample of absorbing exhausts, application of sample needle 22 moves again and deposits the structure sample to reagent to get back to target board platform 21 and continue application of sample

The limiting device comprises limiting switches 271 and limiting members 272, the number of the limiting devices is 3, wherein three limiting members 272 are respectively arranged on the X-direction slider 233, the Y-direction slider 243 and the Z-direction slider 253, three limiting switches 33 are all arranged on the rack, and are respectively arranged at the extreme positions of the X-direction slider 233, the Y-direction slider 243 and the Z-direction slider 253 and are matched with the three limiting members 272 (the matching mentioned here means that the three limiting members 272 can respectively trigger the three limiting switches 271 through movement), the limiting switches 271 and the limiting members 272 are matched to work, when the X-direction slider 233, the Y-direction slider 243 and the Z-direction slider 253 respectively move to the positions of the 3 limiting members 272, the movement is stopped, and the XYZ-direction slider is prevented from continuously moving to cause instrument damage when moving to the extreme positions.

As shown in fig. 1 to 4, the inactivation device of the present invention comprises: sealed cowling 31, atomizer 32, tail gas treatment bottle 33 and pump, sealed cowling 3 installs on hatch door 12, and atomizer 32 and pump are all installed in the frame, and tail gas treatment bottle 33 installs on installation department 14, and sealed cowling 31 forms the inactivation space with the cooperation of target plate platform 21.

The sealed cowling 31 and the target plate platform 21 contactless leaves the slit, has avoided the mechanical friction between sealed cowling 31 and target plate platform 21, can be by slit supplementary air current simultaneously, avoids directly being discharged by gas outlet 313 after leading to the inactivation fog to get into the inactivation space through air inlet 312 and then disinfecting the problem of inactivation because the air exhaust rate is greater than atomizer 32 air intake rate.

The sealing cover 31 is provided with a notch 311, the outer shell of the sealing cover 31 is provided with an air inlet 312 and an air outlet 313, and the positions of the air inlet 312 and the air outlet 313 are matched with the notch 311 (the matching refers to that when the sealing cover 31 covers the target plate platform 21 to form a sealed space, the positions of the air inlet 312 and the air outlet 313 are opposite to the notch 311).

The deactivation liquid is equipped with in the atomizer 32, and the deactivation liquid of this embodiment is the alcohol, and atomizer 32 sets up in the side of target plate platform 21, is connected with air inlet 312, and atomizer 32 adopts ultrasonic atomization alcohol, is provided with liquid level sensor in it and is used for surveying liquid level height, and liquid level sensor transmits liquid level height information to control system to realize automatic liquid feeding function.

Tail gas treatment bottle 33 is connected with gas outlet 313, and the water is equipped with in the bottle, utilizes the very easy characteristic of dissolving in water of alcohol to collect alcohol tail gas, avoids a large amount of alcohol to lose in the air, adopts the lower water of price to collect alcohol tail gas and has not only reduced instrument use cost, more does benefit to the environmental protection simultaneously.

The first diaphragm pump 34 is connected to the tail gas treatment bottle 33 for providing power to drive the alcohol mist to be discharged into the tail gas treatment bottle 33 through the sealing gap 311 and the gas outlet 313. As the mist in the inactivation space is pumped out, negative pressure is generated in the inactivation space, and the mist generated by the atomizer 32 is sucked into the inactivation space, so that the mist in the inactivation space is supplemented.

A second diaphragm pump 35 may also be provided, which is connected to the atomizer 32 and is used to provide power to drive the alcohol mist generated by the atomizer 32 to move into the inactivation space through the air inlet 312 and the sealing cover notch 311. The second diaphragm pump 35 can increase the speed of mist entering the inactivation space, and can increase the inactivation effect. Whether the first diaphragm pump 35 is arranged or not can be selected according to actual requirements.

The deactivation device working process is as follows:

(1) putting the target plate into the target plate platform 21, closing the hatch 12, and driving the sealing cover 31 to move by the hatch 12 to cover the target plate platform 21 to form an inactivation space;

(2) the atomizer 32 atomizes alcohol by using ultrasonic waves to generate alcohol mist, the second diaphragm pump 35 provides power to drive the alcohol mist to move into the inactivation space through the air inlet 312 and the sealing cover notch 311, and the first diaphragm pump 34 provides power to drive the alcohol mist in the inactivation space to move into the tail gas treatment bottle 33 through the sealing cover notch 311 and the air outlet 313;

the first diaphragm pump 34 and the second diaphragm pump 35 are matched with the atomizer 32, the tail gas treatment bottle 33, the sealing cover 31 and the like, so that the alcohol mist airflow in the small space is controlled, and the alcohol is uniformly distributed on the target point, so that the target plate is inactivated. The tail gas treatment bottle 33 is arranged to treat the alcohol tail gas, so that when the alcohol mist soaks each target point, a large amount of alcohol is prevented from being dissipated into the air, and the safety and the use rate of the alcohol are improved.

As shown in fig. 2, 5 and 6, the drying and storing device of the present invention includes: bracket 41, sealing plate 42, vacuum pump 43, first switch 44, second switch 45 and cam mechanism.

The bracket 41 is installed on the frame, the sealing plate 42 and the cam mechanism 43 are both installed on the bracket 41, the bracket 41 is provided with an opening 411, the position of the opening 411 corresponds to the center of the sealing plate 42, and when the target plate platform 21 moves below the sealing plate 42, the sealing plate 4 moves downwards to cover the target plate platform 21 to form a sealed space.

Have dovetail groove, screw hole on the sealing plate 42, still include spring 421 and threaded rod 422, the dovetail groove is used for installing the O circle, and the screw hole is used for the installation to twist directly inserting joint 423 soon, and spring 421 and threaded rod 422 are all four, and the sealing plate 42 passes through threaded rod 422 to be installed on support 41, and the cover is equipped with spring 421 on the threaded rod 422, and the elasticity of spring 421 compresses tightly sealing plate 42 on support 41.

The oil pump 43 is used as a vacuum pump, and the components to be dried include water (saturated vapor pressure at room temperature is approximately 3100Pa), formic acid (saturated vapor pressure at room temperature is approximately 5330Pa), and alcohol (saturated vapor pressure at room temperature is approximately 8500Pa), and the ultimate vacuum that the oil pump 43 can achieve is below 10Pa, which can accelerate the drying speed of the target plate. One connector of a first switch 44 (a two-position two-way electromagnetic valve) is connected with the air suction port of the oil pump 43 through a hard Teflon pipe (a hard pipe for preventing the pipeline from being flattened by atmospheric pressure during air suction), and the other connector is connected with the atmosphere; one connecting port of a second switch 45 (a two-position two-way electromagnetic valve) is connected with the atmosphere, the other connecting port is connected with a three-way joint, the other two ports of the three-way joint are connected with a quick-screwing straight-inserting joint 423 of the sealing plate 42 and an air outlet of the oil pump 43. When the vacuum is pumped, the first switch 44 is opened, and the second switch 45 is closed; when air is discharged, the second switch 45 is turned on, and the first switch 44 is turned off; during storage, the first switch 44 and the second switch 45 are both turned off.

The cam mechanism includes a cam 461, a drying motor 462, a cam holder 463 and a sensor, the cam holder 463 is arranged on the bracket 41, on which the cam 461 and the drying motor 462 are mounted, the cam 461 is in an oval shape and is positioned to fit with the opening 411 on the bracket 41 (the fitting means here that the cam 461 is positioned right above the bracket opening 411), the cam 461 passes through the opening 411 and pushes the sealing plate 42 downward when the cam 461 rotates, so that the O-ring on the sealing plate 42 is pressed on the target plate platform 21 to form a sealed space, and furthermore, in order to prevent the deviation of the rotation angle of the drying motor 462 within a certain range, the cam 461 is provided with a large remote angle of repose (about 30 °). The drying motor 462 is directly connected to the cam 461 to provide a larger torque and a slower speed, thereby making it easier to control the angle of rotation. The sensor includes a first inductive switch 464 and a first trigger piece 465, in this embodiment, the first inductive switch 464 is a photoelectric switch, wherein the first trigger piece 465 is disposed on the cam 461, the first inductive switch 464 is disposed on the cam mounting bracket 463, when the cam 461 is driven by the drying motor 462 to press the sealing plate 42 downwards to reach a maximum stroke (the maximum stroke refers to a position where the edge of the cam is farthest away from the rotation center), the first trigger piece 465 on the cam 461 is adapted to the first inductive switch 464, and the open-close state of the sealed space is determined by the cooperation of the first inductive switch 464 and the first trigger piece 465.

The working process of the target plate drying and storing device is as follows:

(1) after the sample application instrument is powered on, a target plate after sample application is put into the target plate platform 21, the sample application instrument can start a drying function according to an instruction, and the target plate platform 21 moves to the position below the sealing plate 42 under the driving of the Y-direction movement assembly;

(2) the cam 461 is driven by the drying motor 462 to rotate, when the cam 461 moves to be vertical to the plane of the sealing plate 42, the cam 461 penetrates through the opening 411 to press the sealing plate 42 downwards to reach the maximum stroke, the sealing plate 42 and the target plate platform 21 form a sealed space, at the moment, the trigger piece 465 cuts off a signal of the induction switch 464, and the control system judges that the sealed space is in a closed state;

(3) the control system controls the oil pump 43 to work, the first switch 44 is turned on, the second switch 45 is turned off, air in the sealed space is pumped out to form a vacuum sealed space, and the target plate is dried;

(4) after the target plate is dried, the control system further controls the first switch 44 to be closed, the second switch 45 to be opened, air enters the sealed space to reach air pressure balance, the cam 461 rotates under the driving of the drying motor 462 until the longest radius line deviates from the vertical line of the sealing plate 42, the sealing plate 42 is pushed to the support 41 under the action of the spring 421, the first induction switch 464 returns to be light-on, and the control system judges that the sealed space is in an open state;

(5) when the target plate needs to be stored in the dry storage device, the control system controls the first switch 44 and the second switch 45 to be turned off.

(6) The target plate platform 21 moves to the zero position under the drive of the Y-direction movement assembly, and the target plate is taken out.

As shown in fig. 7, the reagent storage device includes: the reagent storage mechanism 51 is provided with one or more reagent holes for storing reagents, and the reagent holes are used for storing reagents such as formic acid, matrix and the like. The reagent sealing motor can rotate the cover 54 to cover the opening of the reagent storage structure 51.

The connecting structure 53 further comprises a second inductive switch 531 and a second trigger piece 532, the second inductive switch 531 is fixed on the frame, and the position of the second inductive switch 531 is matched with the second trigger piece 532. The second triggering piece 532 moves along with the movement of the connecting structure 53, so that the second triggering piece 532 can trigger the second sensing switch 531. The second inductive switch 531 in this embodiment is a photoelectric switch.

The number of the second inductive switch 531 and the second trigger piece 532 in this embodiment is two, and the positions of the second inductive switch 531 and the second trigger piece 532 correspond to each other one by one, and two positions can be detected, where the two positions are the position where the cover 54 is completely closed and the position where the cover is completely opened. Two No. two inductive switch 531 set up side by side, and two No. two trigger piece 532 are located the upper and lower both sides of connection structure 53 respectively, and such structure can make things convenient for inductive switch's installation to just can detect the position after connection structure 53 rotates 180 under the very little condition of occupation space.

A rubber pad is attached to the surface of the cover 54 facing the opening of the reagent storage structure 51, and the sealing ability is improved by the elasticity of the rubber pad at the time of sealing.

The axis of the rotating shaft of the reagent sealing motor in the embodiment is in the horizontal direction, when the reagent needs to be stored in a sealing mode, the cover 54 rotates along the vertical plane, and the cover 54 which falls vertically can be better in sealing performance by means of the gravity of the cover 54. And the lid cuts into and engages the upper surface of the reagent storage structure 51 at an angle. This way, the problem of poor sealing caused by the fact that the cover 54 continues to move after contacting with the upper surface of the reagent storage structure 51 is avoided, abrasion of the cover 54 or a rubber pad can be avoided, and the service life is prolonged.

When the cover 54 is completely covered, a signal of the second induction switch 531 is cut off by the second trigger piece 532, the other second induction switch 531 keeps on light, and the received signal is fed back to the control system, so that the sample application needle 22 is controlled to keep a static state and cannot move downwards; when the cover 54 is completely opened, i.e. rotated by 180 °, the signal of the other second inductive switch 531 is cut off by the other second trigger piece 532, and the control system controls the sample application needle 22 to move downwards to perform the sampling operation; when the cover 54 is in a state other than the above two states, the signals of the two second inductive switches 531 are both on, and the print needle 22 is kept from moving, preventing the print needle 22 from colliding with the cover 54.

A large number of experiments show that the reason that the sample application precision is not high includes factors such as the concentration of the reagent, unstable components, dosage accuracy and the like besides the sample application position.

In the manual sample application process or the existing sample application instrument, the storage of sample application reagents is not taken into account, and only a working position for placing the sample application reagents is provided. Since the substrate components are volatile, the substrate can be deteriorated and cannot be used. The amount of each sample application is very small (1 microliter or less per target), and a small bottle of reagent is usually used for a long time, so that effective sealing is important for ensuring the sample application quality and saving the reagent. The reagent storage device disclosed by the invention can seal the reagent, and effectively slows down the volatilization and deterioration of the reagent. The cover 54 is completely opened when sampling is needed, and the cover 54 is closed when sampling is not needed, so that the volatilization of the reagent is prevented, and the deterioration of the reagent due to the volatilization is avoided.

As shown in fig. 8, the needle washing device includes: the washing tank 61, the needle washing tank 61 has side wall and bottom wall, its side wall is cylindrical, the bottom wall is the toper, offer a inlet 611 on the side wall of the needle washing tank 61, offer a drain port 612 on the bottom wall, the length of the inlet is more than 1mm, the axial deviation of the inlet is from the central line of the needle washing tank.

The liquid inlet peristaltic pump is connected with the needle washing groove 61 and the atomizer 32 and used for adding alcohol to the atomizer 32 and the needle washing groove 61, and the liquid discharge peristaltic pump is connected with a liquid discharge port 612 of the needle washing groove 61 and used for discharging waste liquid which is discharged into a waste liquid bottle.

The liquid inlet peristaltic pump and the liquid discharge peristaltic pump are both arranged on the shell 11, one end of the liquid inlet peristaltic pump is positioned in the operation space, and the other end of the liquid inlet peristaltic pump is positioned in the machine space; one end of the liquid discharge peristaltic pump is positioned in the operation space, and the other end of the liquid discharge peristaltic pump is positioned in the machine space; the liquid inlet peristaltic pump is connected with the liquid inlet 611, and the liquid discharge peristaltic pump is connected with the liquid discharge port 612. Because the service life of the hose in the peristaltic pump is short, the installation mode can not only play a good protection role for the peristaltic pump, but also can conveniently replace the hose of the peristaltic pump in an operation space.

Since the liquid inlet 611 is disposed offset from the central axis of the needle washing groove 61 (i.e. the extension line of the axis of the liquid inlet 611 does not intersect with the central axis of the needle washing groove 61), a rotating liquid flow is formed in the needle washing groove 61 during liquid inlet, which is beneficial to cleaning the outer wall of the sampling needle 22.

When the cleaning of the outer wall is completed and the alcohol overflows the needle washing groove 61, the sample application needle 22 pumps the cleaning liquid in the needle washing groove 61 under the driving of the injection pump to clean the inner wall of the sample application needle 22. After the inner wall is cleaned, the drain port 612 is opened, and the waste liquid is drained into a waste liquid collecting device, preferably an alcohol bottle in this embodiment, by using a drain peristaltic pump.

The needle washing groove 61 is formed in the reagent storage structure 51, and space saving is facilitated.

The working process of the spotting instrument of the invention is as follows:

(1) loading: the hatch door 12 is opened, the target plate loaded with the sample in advance is placed in the target plate platform 21, the hatch door 12 is closed, and simultaneously the sealing cover 31 is driven to move to cover the target plate platform 21 to form a disinfection space,

(2) inactivation: the inactivation device works, and the microorganisms on the target plate are inactivated according to the method;

(3) adding a lysis solution: the Y-direction motion component drives the target plate to move in the Y direction, the X-direction motion component and the Z-direction motion component respectively drive the sample application needle 21 to move in the X direction and the Z direction, so that the sampling and sample application processes are realized, and lysis solution is added to each target spot on the target plate;

(4) drying the target plate and cleaning the sample application needle: drying according to the above-mentioned drying principle, and cleaning the lysate on the sample application needle according to the above-mentioned drying principle;

(5) adding a matrix: adding a substrate to each target point on the target plate according to the same principle of adding the lysis solution;

(6) and (3) drying: drying the target plate with the added matrix according to the drying principle described above;

(5) storing and taking: the target plate can be stored in the sealed space, and can also be driven to move by the Y-direction movement assembly, and the hatch door is opened to take out the target plate.

Thus, a complete sample application workflow is completed.

The above examples are provided only for illustrating the present invention and are not intended to limit the present invention. Changes, modifications, etc. to the above-described embodiments are intended to fall within the scope of the claims of the present invention as long as they are in accordance with the technical spirit of the present invention.

Claims

1. A spotting instrument comprises a frame and a spotting device, and is characterized in that: the sample application device, the drying and storing device and the reagent storing device are all arranged on the frame;

2. The spotting instrument of claim 1 wherein the frame comprises an enclosure enclosing an intra-machine space, a hatch rotationally coupled to the enclosure enclosing an operating space with the enclosure, the inactivation device, the reagent storage device being located in the operating space, and the dry storage device being located in the intra-machine space;

the cabin door is provided with an observation window.

3. Spotting instrument according to claim 1 characterized in that said driving means comprise an X-movement assembly, a Y-movement assembly and a Z-movement assembly;

4. Spotting apparatus according to claims 1-3 characterized in that the spotting device further comprises a positioning device comprising a positioning switch and a positioning plate provided with a plurality of protrusions and a plurality of recesses, the positioning switch and the positioning plate being adapted in position.

5. Spotting apparatus according to claim 4 characterized in that the protrusions and depressions of the positioning plate are evenly spaced and the positioning plate is saw-toothed.

6. The spotting instrument of claim 5 wherein the positioning plates are distributed along the Y-axis and the positioning switches are disposed on a Y-direction slide; or the positioning plates are distributed along the X direction, and the positioning switches are arranged on the X-direction sliding blocks.

7. Spotting apparatus according to claim 1 characterized in that: the sample application device further comprises a limiting device, the limiting device comprises a limiting part and a limiting switch, the limiting part is fixed on the X-direction sliding block and/or the Y-direction sliding block and/or the Z-direction sliding block, and the limiting switch is fixed on the X-axis and/or the Y-axis and/or the Z-axis and matched with the limiting part.

8. The spotting instrument of claims 1-3 and 5-7 wherein the inactivation device comprises a sealing cover for covering the target plate platform, an atomizer and a pump, the sealing cover is mounted on the hatch, the atomizer and the pump are both mounted on the frame, the sealing cover and the target plate platform cooperate to form an inactivation space, and the wall of the inactivation space is provided with an air inlet and an air outlet; the atomizer is equipped with the inactivation liquid in to can the atomizing inactivation liquid, the pump is used for providing power drive inactivation fog and discharges through the gas outlet after the air inlet moves to the inactivation space.

9. Spotting apparatus according to claim 8 characterized in that the inactivation space formed by the sealing cap cooperating with the target plate platform is slit.

10. The spotting instrument of claim 9 wherein the target plate inactivation unit further comprises a tail gas treatment bottle connected to the gas outlet for collecting the inactivation mist exhausted through the inactivation space; the pump is connected with the tail gas treatment bottle and is used for providing power to drive the inactivation mist in the inactivation space to be discharged into the tail gas treatment bottle through the gas outlet;

11. The spotting instrument of claims 1-3, 5-7, 9-10 wherein the dry storage device comprises a support, a sealing plate, a vacuum pump and a cam mechanism, the target plate platform and the support are disposed on the base, the sealing plate, the vacuum pump and the cam mechanism are disposed on the support, the support is provided with an opening corresponding to the cam mechanism, the cam mechanism acts on the sealing plate, the sealing plate and the target plate platform form a sealed space, and the vacuum pump is used for pumping air in the sealed space.

12. Spotting instrument of claim 11 wherein the cam structure comprises a cam, a drying motor and a cam mount, the cam and drying motor being disposed on the cam mount, the cam being connected to a rotating shaft of the drying motor;

the cam is provided with a remote angle of repose of 20-40 degrees;

a bearing is arranged at the maximum stroke position of the cam;

13. The spotting instrument of claims 1-3, 5-7, 9-10, and 12 wherein the reagent storage device comprises a reagent storage structure mounted on the frame, a reagent sealing motor, a connecting structure connected with the reagent sealing motor in a matching manner, and a cover connected with the connecting structure, and the reagent storage structure is provided with one or more reagent holes for storing reagents.

14. Spotting apparatus according to claim 13 wherein the direction of movement of the lid is at an angle to the upper surface of the reagent storage device;

15. The spotting instrument of claims 1-3, 5-7, 9-10, 12, 14-15, wherein the needle washing device comprises a washing tank, a liquid inlet peristaltic pump, and a liquid outlet peristaltic pump, the needle washing tank has a side wall and a bottom wall, a liquid inlet is disposed on the side wall, a liquid outlet is disposed on the bottom wall, the liquid inlet peristaltic pump and the liquid outlet peristaltic pump are both mounted on the housing, one end of the liquid inlet peristaltic pump is located in the operation space, and the other end of the liquid inlet peristaltic pump is located in the machine space; one end of the liquid discharge peristaltic pump is positioned in the operation space, and the other end of the liquid discharge peristaltic pump is positioned in the machine space; the liquid inlet peristaltic pump is connected with the liquid inlet, and the liquid discharge peristaltic pump is connected with the liquid discharge port.

16. The spotting apparatus of claim 15 wherein the length of the loading port is 1mm or more, the extension of the axis of the loading port is deviated from the central axis of the needle washing bath, the cross section of the inner wall of the sidewall is circular or elliptical, and the bottom wall is tapered.