CN115962995A - Full-automatic microbial staining film-making device - Google Patents
Full-automatic microbial staining film-making device Download PDFInfo
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- CN115962995A CN115962995A CN202310251615.0A CN202310251615A CN115962995A CN 115962995 A CN115962995 A CN 115962995A CN 202310251615 A CN202310251615 A CN 202310251615A CN 115962995 A CN115962995 A CN 115962995A
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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Abstract
The invention discloses a full-automatic microbial staining film-making device, which comprises a loading module, wherein an electric objective table is arranged in the loading module, and a microfluidic chip is loaded on the electric objective table; the fluid control module controls the flow state of the sample in the microfluidic chip; the micro-fluidic chip is provided with a sample inlet, a reagent inlet, an oil phase inlet, a dropping liquid port and a waste liquid outlet, and the waste liquid outlet is connected with a waste liquid bottle through a pipeline; this full-automatic microbial staining film-making device can be with micro-fluidic chip's form, the complicated biological sample that probably contains the microorganism dilutes, the schizolysis, the dyeing, hatch, the realization automated processing such as dyeing, form the microorganism sample piece that can supply fluorescence or detect under the bright field environment, the realization is to the automatic quantitative determination of full process of microorganism content, the loaded down with trivial details of professional operating personnel manual operation has been avoided, detection efficiency has both been improved, also avoided manual operation to cause secondary pollution's possibility, improve the accuracy of testing result.
Description
Technical Field
The invention relates to the technical field of microbial staining and detection, in particular to a full-automatic microbial staining film-making device.
Background
At present, microbial staining is finished by a slide making and a set of staining operation procedures, basic procedures of staining relate to slide making, fixing, mordant staining, decoloring, counterstaining, washing, drying and microscopic examination, the staining procedures are generally convenient for manual operation and can only be carried out open or semi-open operation treatment, the risk of pollution of the prepared sample is increased due to the opening of the operation environment, the defects of high process cost, low efficiency, time consumption and the like exist, the quality and the efficiency of the prepared slide are dependent on the proficiency of manual operation, and the sample amount and the reagent amount are not saved.
A syringe pump (US 20090097995 A1) disclosed in the prior art, which applies a linear motion of a piezoelectric linear motor while moving a piston using a linear actuator so that suction and discharge of fluid can be more accurately performed by controlling power supply with respect to the piezoelectric linear motor, includes a cylinder including a receiving space; a piston installed in the cylinder to pump the liquid or powder into and out of the cylinder; and a piezoelectric linear actuator that moves the piston in a reciprocating manner, and further, the piezoelectric linear actuator includes a piezoelectric linear motor; however, the problem of poor stability and sealing performance of the sealing part during the suction process still exists, and therefore, a fully automatic microbial staining slide making device is needed to solve the technical problem.
Disclosure of Invention
The invention aims to provide a full-automatic microbial staining film-making device, which solves the problems of open operation, complex and complicated steps, poor stability and sealing performance of a sealing part in the suction process of a solution and poor heat exchange effect between a heat dissipation part and the surrounding environment in the conventional microbial staining process.
In order to achieve the purpose, the invention provides the following technical scheme: a full-automatic microbial staining slide-making device comprises a loading module, wherein an electric objective table is arranged in the loading module, and a microfluidic chip is loaded on the electric objective table; the fluid control module controls the flow state of the sample in the microfluidic chip; the micro-fluidic chip is provided with a sample inlet, a reagent inlet, an oil phase inlet, a dropping liquid port and a waste liquid outlet, and the waste liquid outlet is connected with a waste liquid bottle through a pipeline.
The full-automatic microbial staining film-making device can realize automatic treatment such as dilution, cracking, staining, incubation and staining of complex biological samples possibly containing microbes in the form of a microfluidic chip, form microbial sample films for detection in a fluorescent or bright field environment, and realize full-process automatic quantitative detection of the content of the microbes.
Specifically, the present apparatus processes a sample including: operations such as sample filtration, cell activation, cell marking, fluorescence scanning, optical microscope detection and the like are integrated into a micro-fluidic chip carrier to be completed, the control of the flow direction and the flow speed of liquid is realized by driving a fluid control module, so that the shunting regulation and control of sample liquid, an activating agent and a coloring agent are realized, the automatic processing of enrichment, activation and dyeing of a sample is realized under a totally-enclosed condition, the complexity of manual operation of professional operators is avoided, the detection efficiency is improved, the possibility of secondary pollution caused by the manual operation is also avoided, the waste of the sample is avoided, and the accuracy of a detection result is improved.
Preferably, this full-automatic microbial staining film-making device, still include, reagent selection module; a reagent table is arranged in the reagent selection module, and a sample solution pipe, an oil phase solution pipe and a plurality of reagent solution pipes are placed on the reagent table.
Preferably, the reagent selection module is provided with a six-position directional valve, the plurality of reagent solution tubes are connected with the six-position directional valve, the six-position directional valve is connected with a liquid inlet tube of the third injection pump, the reagent inlet is connected with a liquid outlet tube of the third injection pump, and the six-position directional valve is electrically connected with the micro control module.
According to the full-automatic microbial staining film-making device, each injection pump can be controlled through the micro-control module, so that the addition amount, the addition speed and the addition pressure of liquid in a corresponding sample solution pipe, an oil phase solution pipe and a reagent solution pipe can enter the micro-fluidic chip for mixed preparation through the sample inlet, the reagent inlet and the oil phase inlet under preset parameters, and thus target liquid drops can be generated, and the film-making efficiency is improved; the micro-control module can also control the opening, closing, conversion and other operations of any valve port in the six-position reversing valve, so that reagent solution is extracted from a plurality of different reagent solution pipes and is injected into the micro-fluidic chip through the reagent inlet in sequence, the addition of the reagent solution in the micro-fluidic chip is continuous, the reagent solution pipes do not need to be replaced for many times, and the tabletting efficiency is improved.
Preferably, the full-automatic microbial staining and slide-making device further comprises a liquid drop sorting module, a microscopic imaging control module and a micro-control module; the micro-imaging control module is electrically connected with the loading module, the fluid control module, the reagent selection module, the droplet sorting module and the micro-imaging control module.
Preferably, the droplet sorting module has a sorting laser illuminator, a Sipm silicon photomultiplier, a charge ring, and a deflection plate arranged in cooperation with each other.
Preferably, a PC host is connected between the microscopic imaging control module and the micro control module, the microscopic imaging control module includes a microscope, and an imaging laser irradiator, an LED fluorescent light source, a fluorescent camera, a silicon photomultiplier and a reflector assembly are cooperatively disposed under the microscope.
According to the full-automatic microbial staining film-making device, a PC (personal computer) host is connected between an image processing module and a micro-control module, a micro-fluidic chip is scanned line by using laser with a specific wavelength, a positive fluorescence position is found and calibrated, then a 400-1000X optical microscope in a micro-imaging control module performs microscopic examination on the positive fluorescence position to quickly obtain a digital image of a bacteria morphological structure, and the shot image is stored, recorded and analyzed in real time through the PC host, so that a real-time detection result is obtained, and a professional can conveniently perform data observation and analysis.
Preferably, a first injection pump, a second injection pump and a third injection pump are arranged in the fluid control module, the first injection pump, the second injection pump and the third injection pump are respectively provided with a liquid inlet pipe and a liquid outlet pipe, a sample solution pipe is connected with the liquid inlet pipe of the first injection pump, a sample inlet is connected with the liquid outlet pipe of the first injection pump, an oil phase solution pipe is connected with the liquid inlet pipe of the second injection pump, and an oil phase inlet is connected with the liquid outlet pipe of the second injection pump.
Preferably, the liquid outlet pipes of the first injection pump, the second injection pump and the third injection pump are electrically connected with a liquid pressure sensor through detection connecting wires, and the liquid inlet pipe and the liquid outlet pipe are respectively provided with an electromagnetic valve.
Preferably, the first injection pump, the second injection pump and the third injection pump are all provided with bottom plates, heat dissipation seats are arranged on the bottom plates, stepping motors are arranged on the heat dissipation seats, screw rods are connected to the output ends of the stepping motors, pushing platforms are arranged on the screw rods, fixing platforms are arranged on the bottom plates, and one end portions of the screw rods are connected to the fixing platforms through shafts.
Preferably, the fixed table is provided with an injection base body, the injection base body is provided with a push rod, one end of the push rod is arranged in the injection base body, the other end of the push rod is connected with the push table, one end of the push rod is fixedly connected to the push table, the end, located in the injection base body, of the push rod is provided with an upper sealing portion, the outlet end, close to the injection base body, of the push rod is provided with a lower sealing portion, the upper sealing portion and the lower sealing portion are movably connected through a connecting convex ball, and the connecting convex ball is a convex structure located in the center of the lower sealing portion.
Preferably, a plurality of annular grooves are formed in the circumferential direction of the side wall of the lower sealing portion, a flow guide channel is formed in the lower sealing portion and communicated with the annular grooves, a conical spring is coaxially arranged at the inner bottom of the injection base body, the conical portion of the conical spring faces the lower sealing portion, and a needle tube is arranged at one end of the injection base body.
This full-automatic microbial staining film-making device, through the design of upper seal portion and lower seal portion, not only strengthened injection base member stability, the leakproofness in the suction process, can avoid the local pressurized of injection base member to lead to its self to take place to break moreover.
Specifically, the upper sealing part and the lower sealing part are preferably made of flexible materials such as rubber, and the lower sealing part is a first sealing component in the injection matrix and needs to continuously bear larger liquid pressure change relative to the upper sealing part, so that the lower sealing part can be arranged in the injection matrix to be long enough to obtain a stronger sealing effect, and accordingly, larger friction resistance can be generated between the sealing component and the inner wall of the injection matrix;
on the other hand, when the push rod performs suction, the lower sealing part is easy to deform, so that liquid is easy to approach one side of the liquid from the lower sealing part, and due to the existence of the flow guide channel in the lower sealing part, a certain accommodating space can be provided between the flow guide channel and the upper sealing part, and the liquid flowing into the flow guide channel due to the deformation of the lower sealing part can be accommodated in the accommodating space or the accommodating space, so that the liquid is prevented from remaining on the inner side wall of the injection base body, further small errors caused by adding reagent solution or sample solution are avoided, and the liquid is prevented from directly leaking onto the push rod after passing through the upper sealing part and the lower sealing part, wherein the inlet end of the flow guide channel is close to the direction of the push rod, and the outlet end is communicated with the annular groove;
on the other hand, the liquid trapped in the containing space formed between the upper sealing part and the lower sealing part prevents the gas from entering from the direction of the push rod, and the sealing effect in the injection base body is ensured;
on the other hand, when the push rod performs suction, as part of gas flows in the accommodating space between the upper sealing part and the lower sealing part through the annular groove and the flow guide channel to a certain extent, the problem that the side wall of the injection matrix is broken due to the fact that the injection matrix is locally pressed too much can be avoided;
on the other hand, through the setting of volute spring, be decomposed into the small bubble when making the bubble that probably exists in the injection in-process liquid through volute spring, reduce its follow-up to dyeing film-making effect and the influence of microscopic examination effect, simultaneously, volute spring's setting has also been solved the lead screw and has excessively promoted the problem that the push rod easily caused the damage to the injection base member, and wherein, volute spring is located between lower sealing and the injection base member bottom surface.
Preferably, the heat dissipation seat is internally provided with a concave platform, the concave platform is provided with a spiral ring, the spiral ring is connected end to end through a flow guide piece, the flow guide piece is of a tubular structure, the inner diameter of the flow guide piece is gradually increased from two ends to the middle, and the outer side wall of the flow guide piece is provided with a rubber pad.
When the full-automatic microbial staining film-making device is used, in order to ensure a sterile environment and avoid open operation, the device is usually placed in a sterile box for operation, and a power mechanism such as a stepping motor is arranged outside the sterile box due to larger heat generation, but the power mechanism is close to the sterile box due to the limitation of a transmission structure, so that the generated heat is easily transferred to the inside of the sterile box, the temperature of a staining film is influenced, and the accurate regulation and control of a microenvironment for bacterial growth in a microfluidic chip in the sample enrichment, activation and staining processes are further influenced; the heat dissipation seat in the device is arranged on the outer side wall of the box plate of the sterile box, so that the stepping motor can be fixedly installed and can also be dissipated, specifically, a liquid inlet and a liquid outlet are formed in opposite side walls of the heat dissipation seat, the liquid inlet and the liquid outlet are respectively communicated with two end parts of the spiral ring, and cooling liquid in the spiral ring inside the spiral ring flows circularly through the circulating pump, so that the heat dissipation purpose is achieved; when the cooling liquid flows through the inner cavity of the flow guide piece, the inner diameter of the cooling liquid is gradually increased from the two ends to the middle, so that the spiral flow track of the cooling liquid is damaged, namely the flow state and the flow speed of the cooling liquid are damaged, the flow time of the cooling liquid in the spiral ring is prolonged, and the heat exchange effect of the cooling liquid and the surrounding environment is improved.
Compared with the prior art, the invention has the beneficial effects that:
1. the full-automatic microbial staining film-making device can realize automatic treatment such as dilution, cracking, staining, incubation and staining of complex biological samples possibly containing microbes in the form of a microfluidic chip, form microbial sample films for detection in a fluorescent or bright field environment, and realize full-process automatic quantitative detection of the content of the microbes.
2. This full-automatic microbial staining film-making device through the design of last sealing and lower sealing, has not only strengthened the stability, the leakproofness of injection base member in suction process, can avoid the local too big emergence of leading to its self to break of injection base member pressurized moreover.
3. This full-automatic microbial staining film-making device through making the coolant liquid when the inside cavity of water conservancy diversion spare, makes the spiral flow path of coolant liquid destroyed, has destroyed its flow state and flow speed promptly to it is long when having prolonged the coolant liquid flow in the spiral ring, has improved its and the heat transfer effect of surrounding environment.
Drawings
FIG. 1 is a schematic structural diagram of a fully automatic microbial staining slide-making device according to a preferred embodiment of the present invention;
FIG. 2 is a process diagram of the full-automatic microbial staining slide-making device provided by the present invention;
FIG. 3 is a schematic structural view of the loading module shown in FIG. 1;
FIG. 4 is a schematic diagram of the droplet sorting module of FIG. 1;
FIG. 5 is a schematic diagram of a configuration of the microscopy imaging control module of FIG. 1;
FIG. 6 is a schematic diagram of the fluid control module of FIG. 1;
FIG. 7 is a schematic diagram of the first syringe pump, the second syringe pump, and the third syringe pump shown in FIG. 6;
FIG. 8 is a schematic structural view of the heat sink shown in FIG. 7;
FIG. 9 is a side view of FIG. 8;
FIG. 10 isbase:Sub>A sectional view taken along line A-A of FIG. 9;
FIG. 11 is a schematic view of the spiral ring of FIG. 10;
FIG. 12 is a schematic view of the configuration of the baffle of FIG. 11;
FIG. 13 is a schematic view of the inner structure of the flow guide member;
FIG. 14 is a schematic view of the injection matrix of FIG. 7;
FIG. 15 is a front view of the injection matrix;
FIG. 16 isbase:Sub>A sectional view taken along line A-A of FIG. 15;
fig. 17 is an enlarged view of i-i in fig. 16.
In the figure: 1. a loading module; 11. an electric stage; 12. a microfluidic chip; 121. a sample inlet; 122. a reagent inlet; 123. an oil phase inlet; 124. a drip port; 125. a waste liquid outlet; 13. a waste liquid bottle; 14. a PC host; 2. a fluid control module; 21. a first syringe pump; 211. a liquid inlet pipe; 212. a liquid outlet pipe; 22. a second syringe pump; 23. a third syringe pump; 24. a base plate; 241. a fixed table; 25. a heat sink; 251. a concave platform; 252. a helical ring; 253. a flow guide member; 254. a rubber pad; 255. a circulation pump; 256. a liquid inlet; 257. a liquid outlet; 26. a stepping motor; 261. a screw rod; 27. a pushing table; 28. injecting a matrix; 281. a push rod; 282. an upper sealing part; 283. a lower seal portion; 284. connecting the convex balls; 285. a ring groove; 286. a flow guide channel; 287. a conical spring; 288. a needle tube; 29. a boxboard; 3. a reagent selection module; 31. a reagent table; 32. a sample solution tube; 33. an oil phase solution pipe; 34. a reagent solution tube; 35. a six-position reversing valve; 36. detecting a connecting line; 37. a liquid pressure sensor; 38. an electromagnetic valve; 4. a droplet sorting module; 41. sorting the laser irradiators; 42. a Sipm silicon photomultiplier; 43. a charge ring; 44. a deflection plate; 5. a microscopic imaging control module; 51. a microscope; 52. an imaging laser irradiator; 53. an LED fluorescent light source; 54. a fluorescence camera; 55. a silicon photomultiplier tube; 56. a mirror assembly; 6. and a micro control module.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Referring to fig. 1 and fig. 3, an embodiment of the present invention includes: a full-automatic microbial staining slide-making device comprises a loading module 1, wherein an electric objective table 11 is arranged in the loading module 1, and a micro-fluidic chip 12 is loaded on the electric objective table 11; the fluid control module 2 is used for controlling the flow state of the sample in the microfluidic chip 12; the microfluidic chip 12 is provided with a sample inlet 121, a reagent inlet 122, an oil phase inlet 123, a dropping port 124 and a waste liquid outlet 125, and the waste liquid outlet 125 is connected with the waste liquid bottle 13 through a pipeline.
This full-automatic microbial staining film-making device can be with micro-fluidic chip 12's form, and the complicated biological sample that probably contains the microorganism dilutes, the realization automated processing such as schizolysis, dyeing, incubate, dyeing, forms the microorganism sample piece that can supply to detect under fluorescence or the bright field environment, realizes the automatic quantitative determination of full process to microorganism content.
Specifically, the present apparatus processes a sample including: operations such as sample filtration, cell activation, cell marking, fluorescence scanning, microscope 51 detection and the like are integrated into a carrier of the microfluidic chip 12 to be completed, the control of the flow direction and the flow rate of liquid is realized by driving the fluid control module 2, so that shunting regulation and control of sample liquid, an activating agent and a coloring agent are realized, automatic processing of enrichment, activation and dyeing of a sample is realized under a fully closed condition, the complexity of manual operation of professional operators is avoided, the detection efficiency is improved, the possibility of secondary pollution caused by manual operation is also avoided, the waste of the sample is avoided, and the accuracy of a detection result is improved.
Referring to fig. 1, the full-automatic microbial staining slide-making device further includes a reagent selection module 3; a reagent table 31 is provided in the reagent selection module 3, and a sample solution tube 32, an oil phase solution tube 33, and a plurality of reagent solution tubes 34 are placed on the reagent table 31.
Referring to fig. 1 and 6, the reagent selection module 3 has a six-position directional valve 35, the plurality of reagent solution tubes 34 are connected to the six-position directional valve 35, the six-position directional valve 35 is connected to the liquid inlet tube 211 of the third injection pump 23, the reagent inlet 122 is connected to the liquid outlet tube 212 of the third injection pump 23, and the six-position directional valve 35 is electrically connected to the micro control module 6.
According to the full-automatic microbial staining film-making device, each injection pump can be controlled by the micro-control module 6, so that the addition amount, the addition speed and the addition pressure of liquid in the corresponding sample solution pipe 32, the oil phase solution pipe 33 and the reagent solution pipe 34 can enter the micro-fluidic chip 12 through the sample inlet 121, the reagent inlet 122 and the oil phase inlet 123 for mixed preparation under preset parameters, and thus target liquid drops can be generated, and the film-making efficiency is improved; the micro-control module 6 can also control the opening, closing, switching and other operations of any valve port in the six-position reversing valve 35, so that reagent solution is extracted from a plurality of different reagent solution pipes 34 and injected into the micro-fluidic chip 12 through the reagent inlets 122 in sequence, the addition of the reagent solution in the micro-fluidic chip 12 is continuous, the reagent solution pipes 34 do not need to be replaced for many times, and the tabletting efficiency is improved.
Referring to fig. 1, 4 and 5, the full-automatic microbial staining slide-making device further includes a droplet sorting module 4, a microscopic imaging control module 5 and a micro-control module 6; the liquid drop sorting module 4 is integrated on the microfluidic chip 12, the microscopic imaging control module 5 collects, stores and analyzes target images in the microfluidic chip 12, and the micro control module 6 is electrically connected with the loading module 1, the fluid control module 2, the reagent selection module 3, the liquid drop sorting module 4 and the microscopic imaging control module 5.
The droplet sorting module 4 has a sorting laser illuminator 41, a Sipm silicon photomultiplier 42, a charge ring 43, and a deflection plate 44, which are disposed in cooperation with each other.
The PC host 14 is connected between the microscopic imaging control module 5 and the micro control module 6, the microscopic imaging control module 5 comprises a microscope 51, and an imaging laser irradiator 52, an LED fluorescent light source 53, a fluorescent camera 54, a silicon photomultiplier 55 and a reflector assembly 56 are arranged below the microscope 51 in a matching manner.
According to the full-automatic microbial staining slide-making device, the PC host 14 is connected between the image processing module and the micro control module 6, the micro control chip 12 is scanned line by using laser with a specific wavelength, a positive fluorescence position is found and calibrated, then a 400-1000X optical microscope 51 in the microscopic imaging control module 5 performs microscopic examination on the positive fluorescence position, a digital image of a bacteria morphological structure is rapidly obtained, and the shot image is stored, recorded and analyzed in real time through the PC host 14, so that a real-time detection result is obtained, and a professional can conveniently perform data observation and analysis.
Referring to fig. 1 and 6, a first injection pump 21, a second injection pump 22 and a third injection pump 23 are disposed in the fluid control module 2, each of the first injection pump 21, the second injection pump 22 and the third injection pump 23 has a liquid inlet pipe 211 and a liquid outlet pipe 212, the sample solution pipe 32 is connected to the liquid inlet pipe 211 of the first injection pump 21, the sample inlet 121 is connected to the liquid outlet pipe 212 of the first injection pump 21, the oil phase solution pipe 33 is connected to the liquid inlet pipe 211 of the second injection pump 22, and the oil phase inlet 123 is connected to the liquid outlet pipe 212 of the second injection pump 22.
Referring to fig. 1 and 6, the liquid outlet pipes 212 of the first injection pump 21, the second injection pump 22 and the third injection pump 23 are electrically connected to a liquid pressure sensor 37 through a detection connection line 36, and the liquid inlet pipe 211 and the liquid outlet pipe 212 are respectively provided with an electromagnetic valve 38.
Referring to fig. 7, the first syringe pump 21, the second syringe pump 22, and the third syringe pump 23 each have a bottom plate 24, the bottom plate 24 is provided with a heat sink 25, the heat sink 25 is provided with a stepping motor 26, an output end of the stepping motor 26 is connected with a screw rod 261, the screw rod 261 is provided with a pushing table 27, the bottom plate 24 is provided with a fixing table 241, and an end of the screw rod 261 is coupled to the fixing table 241.
Referring to fig. 7, 14-17, an injection base 28 is disposed on the fixing platform 241, a push rod 281 is disposed on the injection base 28, one end of the push rod 281 is disposed inside the injection base 28, the other end of the push rod 281 is connected to the push platform 27, one end of the push rod 281 is fixedly connected to the push platform 27, an upper sealing portion 282 is disposed at an end of the push rod 281 located inside the injection base 28, a lower sealing portion 283 is disposed near an outlet end of the injection base 28, the upper sealing portion 282 and the lower sealing portion 283 are movably connected by a connecting convex ball 284, and the connecting convex ball 284 is a convex structure located at a center of the lower sealing portion 283.
Referring to fig. 16 and 17, a plurality of annular grooves 285 are circumferentially formed on a side wall of the lower sealing portion 283, a flow guide channel 286 is formed inside the lower sealing portion 283, the flow guide channel 286 is communicated with the annular grooves 285, a conical spring 287 is coaxially arranged on an inner bottom portion of the injection base 28, a conical portion of the conical spring 287 faces the lower sealing portion 283, and a needle tube 288 is arranged at one end portion of the injection base 28.
This full-automatic microbial staining film-making device, through the design of upper seal 282 and lower seal 283, not only strengthened injection base member 28 stability, the leakproofness in the suction process, can avoid the local too big emergence of leading to it self to break of injection base member 28 moreover pressurized.
Specifically, the upper sealing part 282 and the lower sealing part 283 are preferably made of a flexible material such as rubber, and since the lower sealing part 283 is a first sealing part in the injection base body 28 and needs to continuously bear a large liquid pressure change compared with the upper sealing part 282, in order to obtain a strong sealing effect, the lower sealing part 283 is arranged in the injection base body 28 to be long enough, so that a large frictional resistance is generated between the sealing part and the inner wall of the injection base body 28, and the lower sealing part 283 in the device reduces the contact area between the lower sealing part 283 and the inner wall of the injection base body 28 through a circumferential ring groove 285, so that the noise or blockage problem caused by an excessive frictional force between the lower sealing part 283 and the inner wall of the injection base body 28 during the suction action of the push rod 281 is avoided;
on the other hand, during the pumping action of the push rod 281, the lower sealing portion 283 is easy to deform, so that the liquid is easy to approach one side of the liquid from the lower sealing portion 283, and due to the existence of the flow guide channel 286 in the lower sealing portion 283 in the present device, a certain accommodating space can be provided between the lower sealing portion 283 and the upper sealing portion 282, the liquid flowing into the flow guide channel 286 due to the deformation of the lower sealing portion 283 is firstly accommodated in the accommodating space or the accommodating space, so as to avoid the liquid remaining on the inner side wall of the injection base body 28, further avoid the micro error existing when adding the reagent solution or the sample solution, and avoid the liquid from directly leaking out to the push rod 281 over the upper sealing portion 282 and the lower sealing portion 283, wherein, the outlet end of the flow guide channel 286 is close to the inlet end 281 of the push rod, and the outlet end is communicated with the ring groove 285;
on the other hand, the liquid trapped in the accommodation space formed between the upper sealing portion 282 and the lower sealing portion 283 also prevents the entry of gas in the direction of the push rod 281, ensuring the sealing effect inside the injection base 28;
on the other hand, during the suction action of the push rod 281, since a certain flow of partial gas is carried out in the accommodating space between the upper sealing part 282 and the lower sealing part 283 through the ring groove 285 and the flow guide channel 286, the problem that the side wall of the injection matrix 28 is cracked due to the local over-pressure of the injection matrix 28 can be avoided;
on the other hand, through the arrangement of the conical spring 287, air bubbles possibly existing in the liquid during the injection process are decomposed into small air bubbles when passing through the conical spring 287, so that the subsequent influence on the dyeing slide making effect and the microscopic examination effect is reduced, and meanwhile, the arrangement of the conical spring 287 also solves the problem that the lead screw excessively pushes the push rod 281 to easily damage the injection base body 28, wherein the conical spring 287 is positioned between the lower sealing part 283 and the inner bottom surface of the injection base body 28.
Referring to fig. 7-13, a concave platform 251 is disposed inside the heat sink 25, a spiral ring 252 is disposed on the concave platform 251, the spiral ring 252 is connected end to end by a flow guiding member 253, the flow guiding member 253 has a tubular structure, an inner diameter of the flow guiding member 253 gradually increases from two ends to a middle portion, and a rubber pad 254 is disposed on an outer side wall of the flow guiding member 253.
When the full-automatic microbial staining film-making device is used, in order to ensure a sterile environment and avoid open operation, the device is usually placed in a sterile box for operation, and a power mechanism such as a stepping motor 26 is arranged outside the sterile box due to larger heat generation, but the power mechanism is close to the sterile box due to the limitation of a transmission structure, so that the generated heat is easily transferred to the inside of the sterile box, the temperature of a staining film-making is influenced, and the accurate regulation and control of a microenvironment for the growth of bacteria in the microfluidic chip 12 in the sample enrichment, activation and staining processes are influenced; the heat dissipation seat 25 in the device is arranged on the outer side wall of the aseptic box plate 29, so that the stepping motor 26 can be fixedly installed and also can be dissipated, specifically, a liquid inlet 256 and a liquid outlet 257 are arranged on opposite side walls of the heat dissipation seat 25, the liquid inlet 256 and the liquid outlet 257 are respectively communicated with two end parts of the spiral ring 252, and cooling liquid in the spiral ring 252 inside the spiral ring is circulated and flowed through the circulating pump 255, so that the purpose of dissipating heat is achieved; when the cooling liquid flows through the inner cavity of the flow guide 253, the inner diameter of the cooling liquid is gradually increased from the two ends to the middle, so that the spiral flow trajectory of the cooling liquid is damaged, that is, the flow state and the flow speed of the cooling liquid are damaged, the flow time of the cooling liquid in the spiral ring 252 is prolonged, and the heat exchange effect between the cooling liquid and the surrounding environment is improved, wherein the concave table 251 can be used for collecting condensed water generated by the spiral ring 252, and the arrangement of the elastic blocks can keep the structural form of the spiral ring 252, reduce the possibility of deformation of the spiral ring 252, so as to ensure the distance between the rings of the spiral ring 252 and ensure the heat exchange effect.
The working principle is as follows:
referring to fig. 1 and 2, the full-automatic microbial staining slide-making device specifically comprises the following steps:
s1: preparation of the microfluidic chip 12: aiming at the steps of the dyeing process, the microfluidic chip 12 which meets the functions of sample liquid and reagent input, waste liquid discharge and microorganism capture and enrichment is prepared.
S2: assembling an automatic liquid flow driving system: the sample liquid and the reagent are respectively packaged in an injection matrix 28 with specific specifications, the injection matrix 28 is installed in the fluid control module 2, and the liquid in the injection matrix 28 is pushed to be led out by the operation of an injection pump.
S3: assembling the automatic dyeing system: the sample inlet 121, the oil phase inlet 123, and the reagent inlet 122 of the microfluidic chip 12 are connected to the sample solution tube 32, the oil phase solution tube 33, and the reagent solution tube 34, respectively, and the waste liquid outlet 125 is connected to the waste liquid bottle 13.
S4: dyeing process: and operating corresponding injection pumps, sequentially driving the sample liquid, the reagent and the oil phase liquid in the injection matrix 28 to be led into the microfluidic chip 12, realizing the mixing and dyeing reaction of the reagent and the sample liquid in the microfluidic chip 12, realizing the cleaning and discharging of the dyed reagent in the microfluidic chip 12 by using the buffer solution, finally realizing the capture and fixation of the dyed microorganisms in the microfluidic chip 12, closing the corresponding injection pumps, and completing the automatic slide.
S5: microscopic examination: and (3) sending the micro-fluidic chip 12 which is successfully manufactured into the sheet to the fluorescent and bright field environment for detection.
It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein. Any reference sign in a claim should not be construed as limiting the claim concerned.
Claims (10)
1. A full-automatic microbial staining film-making device, which comprises,
the loading module (1), wherein an electric objective table (11) is arranged in the loading module (1), and a microfluidic chip (12) is loaded on the electric objective table (11);
the fluid control module (2), the fluid control module (2) controls the flow state of the sample in the microfluidic chip (12);
the method is characterized in that: the microfluidic chip (12) is provided with a sample inlet (121), a reagent inlet (122), an oil phase inlet (123), a dropping port (124) and a waste liquid outlet (125), and the waste liquid outlet (125) is connected with a waste liquid bottle (13) through a pipeline.
2. The full-automatic microbial staining film-making device of claim 1, wherein: the utility model provides a full-automatic microbial staining film-making device still includes, reagent selection module (3), be equipped with reagent platform (31) in reagent selection module (3), placed sample solution pipe (32), oil phase solution pipe (33) and a plurality of reagent solution pipe (34) on reagent platform (31).
3. A full-automatic microorganism staining slide-making device according to claim 2, characterized in that: the utility model provides a full-automatic microbial staining film-making device still includes, and liquid drop selects separately module (4), micro-imaging control module (5) and micro-control module (6), liquid drop select separately module (4) integrate in on micro-fluidic chip (12), micro-imaging control module (5) are right target image acquisition, storage and data analysis in micro-fluidic chip (12), micro-control module (6) with load electric connection between module (1), fluid control module (2), reagent selection module (3), liquid drop select separately module (4), micro-imaging control module (5).
4. A full-automatic microbial staining slide production device according to claim 3, wherein: be equipped with first syringe pump (21), second syringe pump (22) and third syringe pump (23) in fluid control module (2), first syringe pump (21), second syringe pump (22) and third syringe pump (23) all have feed liquor pipe (211) and drain pipe (212), sample solution pipe (32) with feed liquor pipe (211) of first syringe pump (21) are connected, sample entry (121) with drain pipe (212) of first syringe pump (21) are connected, oil phase solution pipe (33) with feed liquor pipe (211) of second syringe pump (22) are connected, oil phase entry (123) with drain pipe (212) of second syringe pump (22) are connected.
5. A full-automatic microbial staining production device according to claim 4, wherein: the reagent selection module (3) is provided with a six-position directional valve (35), the reagent solution pipes (34) are connected with the six-position directional valve (35), the six-position directional valve (35) is connected with the liquid inlet pipe (211) of the third injection pump (23), the reagent inlet (122) is connected with the liquid outlet pipe (212) of the third injection pump (23), and the six-position directional valve (35) is electrically connected with the micro control module (6).
6. A full-automatic microbial staining production device according to claim 5, wherein: liquid pressure sensor (37) is connected with through detecting connecting wire (36) electric connection between drain pipe (212) of first syringe pump (21), second syringe pump (22), third syringe pump (23), all be equipped with solenoid valve (38) on feed liquor pipe (211), drain pipe (212).
7. The full-automatic microbial staining film-making device of claim 6, wherein: first syringe pump (21), second syringe pump (22), third syringe pump (23) all have bottom plate (24), be equipped with radiating seat (25) on bottom plate (24), be equipped with step motor (26) on radiating seat (25), be connected with lead screw (261) on the output of step motor (26), be equipped with on lead screw (261) and promote platform (27), be equipped with fixed station (241) on bottom plate (24), a tip coupling of lead screw (261) in fixed station (241).
8. The full-automatic microbial staining film-making device of claim 7, wherein: be equipped with injection base member (28) on fixed station (241), be equipped with push rod (281) on injection base member (28), one end of push rod (281) is located the inside of injection base member (28), the other end of push rod (281) with promote platform (27) fixed connection, push rod (281) are located the end in injection base member (28) is equipped with upper seal portion (282), is close to the exit end of injection base member (28) is equipped with lower seal portion (283), upper seal portion (282) with through connecting protruding ball (284) swing joint between lower seal portion (283).
9. A full-automatic microbial staining slide production device according to claim 8, wherein: a plurality of annular grooves (285) have been seted up to the lateral wall circumference of lower sealing (283), water conservancy diversion passageway (286) have been seted up to the inside of lower sealing (283), water conservancy diversion passageway (286) with annular groove (285) intercommunication, the interior bottom of injection base member (28) is coaxial to be equipped with conical spring (287), conical part orientation of conical spring (287) lower sealing (283), a tip of injection base member (28) is equipped with needle tubing (288).
10. The method for using a full-automatic microbial staining production device according to claim 9, comprising the steps of: the method comprises the steps of micro-fluidic chip (12) preparation, automatic liquid flow driving system assembly, automatic dyeing system assembly, dyeing process and microscopic examination.
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