CN212111056U - Droplet reading instrument - Google Patents

Abstract

The utility model discloses a droplet reading appearance, include: the device comprises a liquid storage assembly, a liquid conveying system, a sample loading assembly and an optical detection assembly; the liquid storage assembly is used for providing detection oil and collecting waste oil, and the sample loading assembly is used for sucking sample micro-droplets to be detected placed in the micro-droplet reading instrument and conveying the sample micro-droplets to be detected through the liquid conveying system to enable the sample micro-droplets to be detected to be sequentially arranged and pass through the detection position of the optical detection assembly so as to perform fluorescence detection on the sample micro-droplets to be detected. The utility model can realize the three-dimensional movement of the sample injection needle through the three-dimensional moving mechanical arm so as to perform sample injection; the defect of insufficient sample suction caused by depth errors of the sample storage holes can be overcome by arranging the buffer connecting block, and the sample injection needle can penetrate into the bottom of each sample storage hole to ensure that the sample is completely absorbed; through the optimized liquid path structure, the control difficulty of the system can be reduced in a limited way, and meanwhile, the robustness of the system is improved; the utility model discloses a droplet reading appearance has optimized the structure, and whole platform instrument structural configuration is clear, easy dismounting.

Description

Droplet reading instrument

Technical Field

The utility model relates to a biological detection technical field, in particular to droplet reading appearance.

Background

The high-sensitivity and rapid nucleic acid detection technology has strong advantages in the fields of low-abundance nucleic acid detection such as cancer molecular marker discovery, infectious diseases, genetic disease research and the like, and has great significance for the extreme research on disease onset, early diagnosis and personalized treatment! Typical nucleic acid detection techniques include fluorescence quantitative PCR, molecular hybridization, and gene sequencing, which have limited detection accuracy, high cost, and long time consumption. The Digital PCR (Digital PCR-dPCR) technology is a novel nucleic acid detection and quantification method, and the copy number of a target sequence is directly detected in an absolute quantification mode without depending on a standard curve and a reference sample. Principle of digital PCR: a standard PCR reaction is distributed into a large number of tiny reactors, each reactor contains or does not contain one or more copies of a target molecule (DNA template), so that single-molecule template PCR amplification is realized, and after the amplification is finished, the copy number of a target sequence is counted through the number of positive reactors. The digital PCR process comprises: droplet generation → overlay → amplification → droplet readout.

Wherein droplet generation is used to generate thousands of droplets, implemented using a droplet generator; the film covering is used for covering 96 microdroplet samples in a 96-well plate, so that the samples are prevented from being baked or damaged in the subsequent pcr process, and the film covering is realized by using a film covering machine which is only a simple mechanical heating and gluing process; the amplification is realized by a PCR instrument which is a temperature cycle process essentially and is used for realizing the copy of DNA in a microdroplet sample, and mature PCR amplification instruments such as Edwarder, Langi and the like are available on the market; the microdroplet reading means that microdroplets are arranged in turn and pass through an optical detection area, negative and positive signals of each microdroplet are detected by an optical means, and finally, the concentration of a target gene in an original sample is calculated according to a mathematical calculation formula of Poisson distribution, so that the purpose of detecting the target gene of the sample is realized, and the microdroplet reading can be realized by a reading instrument.

Four instruments are required for the digital PCR process: the device comprises a droplet generator, a film laminator, a PCR (polymerase chain reaction) instrument and a reading instrument, wherein the technical thresholds of the film laminator and the PCR instrument are lower, and the droplet generator and the reading instrument have higher technical thresholds. The reading instrument is used for sequentially arranging the microdroplets in the sample, carrying out fluorescence detection, and calculating the concentration of the original sample through a later data processing algorithm. The existing reading instrument has many defects, such as complex structure of the whole machine, inconvenient use of a device for detecting oil supply and waste oil recovery, low utilization rate of a sample to be detected and the like, so that a more reliable scheme is needed at present.

SUMMERY OF THE UTILITY MODEL

The utility model aims to solve the technical problem that to the not enough among the above-mentioned prior art, provide a droplet reading appearance.

In order to solve the technical problem, the utility model discloses a technical scheme is: a droplet reader comprising: the device comprises a liquid storage assembly, a liquid conveying system, a sample loading assembly and an optical detection assembly;

the liquid storage assembly is used for providing detection oil and collecting waste oil, and the sample loading assembly is used for sucking sample micro-droplets to be detected placed in the micro-droplet reading instrument and conveying the sample micro-droplets to be detected through the liquid conveying system to enable the sample micro-droplets to be detected to be sequentially arranged and pass through the detection position of the optical detection assembly so as to perform fluorescence detection on the sample micro-droplets to be detected.

Preferably, the liquid storage assembly comprises a liquid storage base, a liquid tank box slidably arranged on the liquid storage base, and at least two liquid storage tanks arranged in the liquid tank box; the at least two liquid storage tanks at least comprise a detection oil tank and a waste oil tank;

the sample loading assembly comprises a three-dimensional moving mechanical arm, a sample storage device and a sample injection needle arranged on the three-dimensional moving mechanical arm; the sample storage device is used for storing the micro-droplets of the sample to be detected.

Preferably, the liquid delivery system include with detect oil pipeline, the setting of oil tank intercommunication be in oil pump on the oil pipeline, through the oil inlet with detection chip, the sample pump of the terminal intercommunication of output of oil pipeline, communicate in sample pump and the sample input pipeline between the introduction port that detects the chip, set up and be in the switching-over valve on the sample input pipeline, communicate in inhale the appearance pipeline between switching-over valve and the introduction needle, communicate in detect the waste oil pipeline between total export of chip and the waste oil tank and set up and be in solenoid valve on the waste oil pipeline.

Preferably, the direction valve at least includes a first port, a second port and a third port, and is switchable between a first state and a second state, the first state being: the first port is communicated with the second port, and the second port is communicated with the third port; the second state of disconnection is: the second port and the third port are conducted, and the first port and the second port are disconnected;

the second port is communicated with the sample pump through the sample conveying pipeline, the third port is communicated with the sample inlet of the detection chip through the sample conveying pipeline, and the first port is communicated with the sample injection needle through the sample suction pipeline.

Preferably, the oil inlet of detection chip includes 2, the inside miniflow channel structure that has the cross type of detection chip, the miniflow channel structure is in four sides of detection chip form respectively introduction port, 2 oil inlets and total export, 2 oil inlet symmetric distribution in introduction port both sides.

Preferably, the sample loading device further comprises a bottom plate, and the liquid storage assembly, the liquid conveying system, the sample loading assembly and the optical detection assembly are all arranged on the bottom plate;

the liquid storage base is fixedly connected to the bottom plate, and the liquid storage tank comprises a tank body, a tank opening arranged at the top of the tank body, a joint pipe arranged on the tank opening, a joint converter with the lower end inserted in the joint pipe and the upper end extending out, and a tank cover in threaded fit with the joint pipe and used for fixing the lower end of the joint converter in the joint pipe;

a plurality of channels for communicating the inside of the tank body with the outside are arranged in the joint converter;

the channel comprises a horizontal channel and a vertical channel which are communicated with each other, the horizontal channel forms an external pipeline interface on the side surface of the upper end of the joint converter, and the vertical channel forms an internal pipeline interface on the lower end surface of the joint converter.

Preferably, the middle part of the tank cover is provided with a through hole, and the inner edge of the upper end of the tank cover is provided with an annular flange;

an annular baffle plate is arranged on the periphery of the joint converter, and the outer diameter of the annular baffle plate is smaller than the diameter of the through hole and larger than the inner diameter of the annular flange;

the joint converter is inserted in the joint pipe, and after the tank cover is in threaded connection with the joint pipe, the annular flange is pressed against the upper surface of the annular baffle.

Preferably, the three-dimensional mobile robot arm comprises an X-axis motion assembly arranged on the base plate, a Y-axis motion assembly arranged on the X-axis motion assembly, and a Z-axis motion assembly arranged on the Y-axis motion assembly;

the Z-axis motion assembly comprises a Z-axis mounting plate arranged on the Y-axis motion assembly, a Z-axis motor arranged on the Z-axis mounting plate, a Z-axis screw rod in driving connection with the Z-axis motor and a Z-axis sliding sleeve in threaded fit with the Z-axis screw rod;

the sample injection needle is connected to the Z-axis sliding sleeve through a buffer connecting block.

Preferably, a threaded hole is formed in the buffer connecting block, a boss is arranged at the upper part of the Z-axis sliding sleeve, a through hole is formed in the boss, a screw with the diameter smaller than that of the through hole is inserted into the through hole, the lower part of the screw is fixedly connected with the threaded hole, and a nut with the diameter larger than that of the through hole is arranged at the upper end of the screw; a spring is sleeved on the Z-axis sliding sleeve, the upper end of the spring is in contact with the bottom surface of the boss, and the lower end of the spring is in contact with the buffer connecting block;

the buffer connecting block is provided with a via hole with a diameter larger than that of the Z-axis screw rod for the Z-axis screw rod to pass through, and the upper part of the buffer connecting block is also provided with a holding groove for holding the Z-axis sliding sleeve.

Preferably, a detection pipeline section is connected between the main outlet of the detection chip and the waste liquid pipeline, and the detection pipeline section is positioned at the detection position of the optical detection component;

the optical detection assembly comprises at least two laser light sources, a transmission light path for transmitting light emitted by the at least two laser light sources to a sample droplet to be detected on the detection pipeline section, at least two lateral detectors for collecting fluorescence generated by the excitation of the sample droplet to be detected on the detection pipeline section, and a forward detector for collecting laser passing through the sample droplet to be detected on the detection pipeline section.

The utility model has the advantages that:

the liquid storage component of the utility model can keep the position of the joint converter unchanged basically in the process of unscrewing the tank cover, can not cause the pipeline connected with the joint converter to twist and wind due to the rotation of the tank cover, and is convenient to open the tank cover; the sliding device is arranged at a high position and a low position, so that the space can be saved, the installation is convenient, and the replacement of the liquid storage tank is convenient; by arranging the magnets and the iron sheets, the stability of the liquid tank box after being pushed into the base can be improved; the liquid level in the tank body can be monitored by arranging the liquid level sensor;

the utility model can realize the three-dimensional movement of the sample injection needle through the three-dimensional moving mechanical arm so as to perform sample injection; the defect of insufficient sample suction caused by depth errors of the sample storage holes can be overcome by arranging the buffer connecting block, and the sample injection needle can penetrate into the bottom of each sample storage hole to ensure that the sample is completely absorbed;

the utility model can reduce the control difficulty of the system in a limited way and improve the robustness of the system at the same time through the optimized liquid path structure;

the utility model can realize double fluorescence detection and even more multiple fluorescence detection, thereby greatly improving the utilization rate of the sample;

the utility model discloses a droplet reading appearance adopts the modular design theory, has optimized the structure, and whole platform instrument structure overall arrangement is clear, easy dismounting.

Drawings

FIG. 1 is a schematic diagram of a droplet reading apparatus according to the present invention;

FIG. 2 is a schematic view of another perspective of the droplet reader of the present invention;

FIG. 3 is a schematic structural view of the liquid storage assembly of the present invention;

FIG. 4 is a schematic view of the liquid storage assembly of the present invention from another perspective;

FIG. 5 is a schematic side view of the reservoir assembly of the present invention;

FIG. 6 is a schematic side view of the reservoir assembly of the present invention;

FIG. 7 is a schematic view of the liquid storage tank of the present invention;

FIG. 8 is an exploded view of the liquid storage tank of the present invention;

fig. 9 is a schematic cross-sectional structural view of the joint converter of the present invention;

FIG. 10 is a schematic view of a partial cross-sectional structure of a liquid storage tank of the present invention;

fig. 11 is a schematic view of a liquid path of the liquid delivery system of the present invention;

fig. 12 is a schematic structural diagram of the detection chip of the present invention;

FIG. 13 is a schematic cross-sectional view of the detection chip of the present invention;

fig. 14 is a schematic structural view of the three-dimensional mobile robot arm of the present invention;

fig. 15 is a schematic structural view of another view angle of the three-dimensional mobile robot according to the present invention;

fig. 16 is a schematic structural view of the Z-axis moving assembly and the buffer connecting block of the present invention;

fig. 17 is an exploded view of the Z-axis moving assembly and the buffer connecting block of the present invention;

fig. 18 is a schematic sectional view of the buffering connecting block of the present invention;

fig. 19 is a schematic structural view of the sample storage device of the present invention;

fig. 20 is a schematic sectional view of the sample storage device of the present invention;

fig. 21 is a light path diagram of the optical detection assembly of the present invention.

Description of reference numerals:

1-liquid storage component;

10-liquid storage base; 100-base plate; 101-a first side panel; 102 — a second side plate; 103-installation space; 104-a pillar;

11-liquid container box;

12-a liquid storage tank; 120-a tank body; 121-tank opening; 122 — connector tube; 123-splice converter; 124-tank cover; 125-channel; 127-a liquid level sensor; 128-L shaped anchor tabs; 129-a mounting frame; 1230-ring catch; 1240-through holes; 1241 — annular flange; 1250 — horizontal channel; 1251 — vertical channel; 1252-external line interface; 1253 — internal pipe interface; 1290-horizontal mounting plate; 1291-a high vertical mounting plate; 1292-a low level vertical mounting plate; 1293-mounting a through hole; 1294-baffle plate; 1295-a magnet;

13-a sliding device;

14-detecting the oil tank;

15-a waste oil tank;

2-a liquid delivery system;

20-an oil pipeline; 21-an oil pump; 22-detection chip; 23-sample pump; 24-a sample conveying pipeline; 25-a diverter valve; 26-a sample sucking pipeline; 27-waste oil line; 28-electromagnetic valve; 220-sample inlet; 221-an oil inlet; 222 — total exit; 223-detecting the pipeline section; 250 — a first port; 251 — a second port; 252 — third port;

3-a sample loading assembly;

30-three-dimensional moving mechanical arm;

300-X axis motion assembly; 3000-X axis motor; 3001-X axis lead screw; 3002 — X axis slide; 3003-X-axis optocoupler; 3004-X baffle plate;

301-Y axis motion assembly; 3010-Y axis mounting plate; 3011-Y axis motor; 3012-Y axis lead screw; 3013-Y-axis slide; 3014 — Y-axis optocoupler; 3015-Y stop piece;

302-Z axis motion assembly; 3020-Z-axis mount plate; 3021-Z axis motor; 3022-Z-axis lead screw; 3023-Z-axis sliding sleeve; 3024-boss; 3025-a through hole; 3026-screw; 3027-screw cap; 3028-spring; 3029 — Z-axis optocoupler;

303-mounting a bracket;

31-a sample storage device; 310-orifice plate base; 311-sample storage well plate; 312-a platen;

32-a sample injection needle;

33-buffer connecting block; 330-a via hole; 331-accommodating grooves; 332-Z stop; 333-threaded hole;

4-an optical detection assembly; 40-a first laser; 41-a second laser; 42 — first lateral detector; 43 — second lateral detector; 44 — a transmission optical path; 45-second dichroic mirror; 46-a first filter; 47 — a second filter; 49-forward detector; 440 — a first mirror; 441-a first dichroic mirror; 442-a focusing lens;

5-bottom plate.

Detailed Description

The present invention is further described in detail below with reference to examples so that those skilled in the art can implement the invention with reference to the description.

It will be understood that terms such as "having," "including," and "comprising," as used herein, do not preclude the presence or addition of one or more other elements or groups thereof.

As shown in fig. 1-2, a droplet reader of the present embodiment includes: the device comprises a liquid storage component 1, a liquid conveying system 2, a sample loading component 3 and an optical detection component 4;

stock solution subassembly 1 is used for providing the detection oil and collects waste oil, and the subassembly of getting ready 3 is used for absorbing the sample droplet that awaits measuring of putting into the droplet reading appearance to carry through liquid delivery system 2 and make the sample droplet that awaits measuring arrange in proper order through the detection position of optical detection subassembly 4, in order to carry out fluorescence detection to the sample droplet that awaits measuring. The detection oil is used for promoting the micro-droplets of the sample to be detected to be sequentially arranged to pass through the detection position, so that the fluorescence detection of each micro-droplet is realized. The optical detection component 4 emits laser to irradiate the micro-droplet of the sample to be detected, so that the micro-droplet is excited to generate fluorescence, and the fluorescence signal is collected and subjected to post data processing to realize sample detection.

The liquid storage assembly 1 comprises a liquid storage base 10, a liquid tank box 11 slidably arranged on the liquid storage base 10 and at least two liquid storage tanks 12 arranged in the liquid tank box 11; the at least two liquid storage tanks 12 at least comprise a detection oil tank 14 and a waste oil tank 15;

the sample loading assembly 3 comprises a three-dimensional moving mechanical arm 30, a sample storage device 31 and a sample injection needle 32 arranged on the three-dimensional moving mechanical arm 30; the sample storage device 31 is used for storing the micro-droplets of the sample to be detected.

In one embodiment, referring to fig. 11-13, the liquid delivery system 2 includes an oil pipeline 20 in communication with the detection oil tank 14, an oil pump 21 disposed on the oil pipeline 20, a detection chip 22 in communication with an output end of the oil pipeline 20 through an oil inlet 221, a sample pump 23, a sample pipeline 24 in communication between the sample pump 23 and a sample inlet 220 of the detection chip 22, a reversing valve 25 disposed on the sample pipeline 24, a sample suction pipeline 26 in communication between the reversing valve 25 and the sample needle 32, a waste oil pipeline 27 in communication between a main outlet 222 of the detection chip 22 and the waste oil tank 15, and an electromagnetic valve 28 disposed on the waste oil pipeline 27.

The direction valve 25 includes at least a first port 250, a second port 251, and a third port 251, and can switch between a first state and a second state, where the first state is: the first port 250 and the second port 251 are conducted, and the second port 251 and the third port 251 are conducted; the second off state is: the second port 251 and the third port 251 are connected, and the first port 250 and the second port 251 are disconnected; the second port 251 is communicated with the sample pump 23 through the sample transmission pipeline 24, the third port 251 is communicated with the sample inlet 220 of the detection chip 22 through the sample transmission pipeline 24, and the first port 250 is communicated with the sample injection needle 32 through the sample suction pipeline 26. The diverter valve 25 in this embodiment is a three-position two-way diverter valve 25.

The detection chip 22 can be a microfluidic chip made of transparent PMMA.

The oil inlets 221 of the detection chip 22 include 2, a cross-shaped micro-channel structure is arranged in the detection chip 22, the micro-channel structure forms a sample inlet 220, 2 oil inlets 221 and a total outlet 222 on four side surfaces of the detection chip 22, and the 2 oil inlets 221 are symmetrically distributed on two sides of the sample inlet 220.

The process of sample suction is as follows: the electromagnetic valve 28 is closed, the first port 250 and the second port 251 are communicated, the sample pump 23 works to suck back samples, and the sample droplets to be detected absorbed by the sample injection needle 32 are filled in the sample conveying pipeline 24 after sequentially passing through the sample sucking pipeline 26, the first port 250 and the second port 251;

detection process (sample input into the detection chip 22): the electromagnetic valve 28 is opened, the reversing valve 25 is switched from the first state to the second state, namely the second port 251 and the third port 251 are communicated, and the first port 250 and the second port 251 are disconnected; the oil pump 21 works to push the detection oil to move forward, and the sample pump 23 works to push the micro-droplets of the sample to be detected in the sample transmission pipeline 24 to move forward; in the detection chip 22, the channel 125 has a cross structure, the sample droplets to be detected move forward, and the detection oil cuts the sample droplets to be detected from two vertical sides to disperse the droplets in the sample one by one. Of course, the criss-cross configuration could be replaced with a T-configuration or a Y-configuration, i.e., the test oil would be sheared from only one side. It will be appreciated that a criss-cross configuration should be the most preferred arrangement. The control difficulty of the system can be reduced in a limited way through the optimized liquid path structure,

in one embodiment, referring to fig. 3-10, the device further comprises a housing (not shown) and a bottom plate 5, wherein the liquid storage assembly 1, the liquid delivery system 2, the sample loading assembly 3 and the optical detection assembly 4 are disposed on the bottom plate 5 and are located inside the housing;

the liquid storage base 10 is fixedly connected to the bottom plate 5, and the liquid storage tank 12 comprises a tank body 120, a tank opening 121 arranged at the top of the tank body 120, a joint pipe 122 arranged on the tank opening 121, a joint converter 123 with the lower end inserted into the joint pipe 122 and the upper end extending out, and a tank cover 124 in threaded fit with the joint pipe 122 and used for fixing the lower end of the joint converter 123 in the joint pipe 122;

a plurality of channels 125 for communicating the inside of the tank 120 with the outside are arranged in the joint converter 123;

an L-shaped fixing piece 128 is fixedly connected to the joint converter 123, and the other end of the L-shaped fixing piece 128 is connected to the tank 120.

The passage 125 includes a horizontal passage 1250 and a vertical passage 1251 communicating with each other, the horizontal passage 1250 forming an external pipe joint 1252 at a side of an upper end of the joint converter 123, and the vertical passage 1251 forming an internal pipe joint 1253 at a lower end of the joint converter 123.

The middle part of the tank cover 124 is provided with a through hole 1240, and the inner edge of the upper end of the tank cover is provided with an annular flange 1241; the outer periphery of the adapter converter 123 is provided with an annular catch 1230, and the outer diameter of the annular catch 1230 is smaller than the diameter of the through hole 1240 and larger than the inner diameter of the annular flange 1241; the adapter 123 is inserted into the adapter tube 122, and after the can lid 124 is screwed to the adapter tube 122, the annular flange 1241 presses against the upper surface of the annular flap 1230.

The joint converter 123 is connected with the tank body 120 through the L-shaped fixing piece 128, so that the joint converter cannot rotate, and pipelines connected with the joint converter can be prevented from twisting and winding; and does not fall downward. And the L-shaped fixing piece 128 has a certain elasticity so that the joint converter 123 can vertically move at a small distance. When the cover 124 is tightened, the internal threads of the cover 124 engage the external threads of the adapter tube 122, the cover 124 rotates, the adapter 123 does not rotate, the cover 124 rotates downward, and the annular flap 1230 of the adapter 123 is pressed downward by the annular flange 1241 (the adapter 123 moves slightly downward), thereby securing the adapter 123. Because the tank has limited volume and needs to be replaced periodically, the detection oil is stable detection oil, the waste liquid is cost, the tank and the waste liquid are both special, and the phenomenon of cross contamination can not be sent, so that the cover (comprising the tank cover 124 and the joint converter 123) is not replaced when the liquid storage tank 12 is replaced, and the trouble of detaching the pipeline (the pipeline is connected to the joint converter 123) is avoided. In this embodiment, the position of the adapter converter 123 can be maintained substantially unchanged during unscrewing of the tank cover 124, and the pipeline connected to the adapter converter 123 is not twisted due to rotation of the tank cover 124.

Further, a liquid level sensor 127 is disposed on the tank 120. The level sensor 127 employs a non-contact (e.g., capacitive or inductive) level sensor 127, such as the level sensor 127 model XKC-Y26, for level monitoring within the tank 120. The result monitored by the liquid level sensor 127 can be transmitted to the droplet reading instrument for complete machine display, and a user can be reminded to replace the liquid storage tank 12 in time.

Further, the liquid tank box 11 is connected with the base in a sliding way through a sliding device 13; the bottom of the liquid tank box 11 is fixedly connected with a mounting rack 129, the mounting rack 129 comprises a horizontal mounting plate 1290 fixedly connected with the bottom of the liquid tank box 11, and a high-position vertical mounting plate 1291 and a low-position vertical mounting plate 1292 fixedly connected with two sides of the horizontal mounting plate 1290, and the height of the bottom edge of the high-position vertical mounting plate 1291 is higher than that of the bottom edge of the low-position vertical mounting plate 1292. The liquid storage base 10 comprises a base plate 100, and a first side plate 101 and a second side plate 102 fixedly connected to two sides of the base plate 100, wherein the height of the second side plate 102 is smaller than that of the first side plate 101, and an installation space 103 is left between the upper end of the second side plate 102 and the bottom of the liquid tank 11 so as to facilitate the installation of the sliding device 13; the slide assembly 13 includes a high point disposed between the mounting bracket 129 and the base, one of which is disposed between the high vertical mounting plate 1291 and the first side plate 101 and the other of which is disposed between the low vertical mounting plate 1292 and the second side plate 102.

In the preferred embodiment, the sliding device 13 is a slide rail of a drawer type, one half of which is connected to the base and the other half of which is connected to the mounting frame 129, the two halves being slidable relative to each other. In another embodiment, the sliding device 13 may also be a combination of a sliding block and a sliding rail, that is, the sliding block and the sliding rail are respectively disposed on the liquid storage base 10 and the mounting frame 129, so as to realize the slidable connection between the base and the mounting frame 129.

The liquid storage device of this embodiment is mainly used in a droplet reader, the liquid storage tank 12 needs to be replaced frequently, and in order to save the height space, the liquid storage tank 11 is horizontally drawn out from the side for replacement. The sliding device 13 is arranged at a high position and a low position, so that the space can be saved, and the installation is convenient. Further, a mounting through hole 30251293 is formed in a portion of the lower vertical mounting plate 1292 located in the mounting space 103, and the mounting of the sliding device 13 on the higher vertical mounting plate 1291 can be facilitated through the mounting through hole 30251293.

A baffle 1294 is fixedly connected to the inner end of the horizontal mounting plate 1290, a magnet 1295 is disposed on the baffle 1294, a support 104 is disposed at the inner end of the base, and an iron sheet (not shown in the figure) for attracting the magnet 1295 is disposed on the support 104. After the liquid tank box 11 is pushed into the base, the magnet 1295 is attracted with the iron sheet, so that the liquid tank box 11 is kept relatively fixed in the base.

In one embodiment, referring to fig. 14 to 20, the three-dimensional moving robot 30 includes an X-axis moving assembly 300 disposed on the base plate 5, a Y-axis moving assembly 301 disposed on the X-axis moving assembly 300, and a Z-axis moving assembly 302 disposed on the Y-axis moving assembly 301;

the Z-axis moving assembly 302 comprises a Z-axis mounting plate 3020 arranged on the Y-axis moving assembly 301, a Z-axis motor 3021 arranged on the Z-axis mounting plate 3020, a Z-axis screw 3022 in driving connection with the Z-axis motor 3021, and a Z-axis sliding sleeve 3023 in threaded fit with the Z-axis screw 3022;

the sample injection needle 32 is connected to the Z-axis sliding sleeve 3023 through a buffer connecting block 33.

The sample storage device 31 comprises a well plate base 310, a sample storage well plate 311 arranged on the well plate base 310, and a pressing plate 312 arranged on the sample storage well plate 311. The sample storage hole plate 311 is provided with a plurality of sample storage holes for storing samples, and in this embodiment, a 96-well plate is selected. The hole plate base 310 is provided with a plurality of hole structures matched with the bottoms of the sample storage hole plates 311, and the pressing plate 312 is also provided with a plurality of holes corresponding to the positions and the number of the sample storage holes above the sample storage hole plates 311. The pressing plate 312 may also be provided with a conventional pressing device to press and fix the sample storage plate 311 on the plate base 310 through the pressing plate 312. During detection, the micro-droplets to be detected are stored in the pore plate, then the pore plate is placed in a micro-droplet reading instrument, and then the three-dimensional moving mechanical arm 30 is used for sucking the samples in the pore plate so as to carry out detection.

Wherein, because the existence of sample storage hole 311 machining error, the degree of depth of every sample storage hole probably has the difference, if the sample needle 32 moves to the same height every time and leads to easily storing up the unable absorption of the downthehole sample of appearance, leads to the sample extravagant, and to some precious samples, more need solve this problem. This problem can be solved in this application by the buffer connection block 33.

Specifically, in a preferred embodiment, a threaded hole 333 is formed in the buffer connecting block 33, a boss 3024 is formed in the upper portion of the Z-axis sliding sleeve 3023, a through hole 3025 is formed in the boss 3024, a screw 3026 with a diameter smaller than that of the through hole 3025 is inserted into the through hole 3025, the lower portion of the screw 3026 is fixedly connected with the threaded hole 333, and a nut 3027 with a diameter larger than that of the through hole 3025 is formed in the upper end of the screw 3026; a spring 3028 is sleeved on the Z-axis sliding sleeve 3023, the upper end of the spring 3028 is in contact with the bottom surface of the boss 3024, and the lower end of the spring 3028 is in contact with the buffer connecting block 33; the buffer connecting block 33 is provided with a via hole 330 which is larger than the Z-axis screw rod 3022 in diameter and is used for the Z-axis screw rod 3022 to pass through, the upper part of the buffer connecting block 33 is also provided with a containing groove 331 used for containing the Z-axis sliding sleeve 3023, and the threaded hole 333 is formed downwards from the bottom of the groove.

The working principle is as follows: the Z-axis screw rod 3022 rotates to drive the Z-axis sliding sleeve 3023 to slide up and down, when the Z-axis sliding sleeve 3023 slides down, the Z-axis sliding sleeve 3023 drives the buffer connection block 33 and the sample injection needle 32 thereon to move down together under the action of the spring 3028 and the self-weight of the buffer connection block 33, when the sample injection needle 32 contacts the inner wall of the bottom of the sample storage hole, the Z-axis sliding sleeve 3023 continues to move down by a small distance, when the Z-axis sliding sleeve 3023 continues to move down, the boss 3024 is pressed down to compress the spring 3028, and the upper part of the screw 3026 in threaded connection with the buffer connection block 33 extends out upward relative to the boss 3024. When the limit distance of the downward movement of the Z-axis sliding sleeve 3023 is preset, the Z-axis sliding sleeve 3023 still moves downward a small distance after the sample injection needle 32 contacts the inner wall of the bottom of the sample storage hole, so that even if there is an error in the depth of each sample storage hole, the sample injection needle 32 can penetrate into the bottom of each sample storage hole to ensure that the sample is completely absorbed. On the other hand, after the sample injection needle 32 contacts the inner wall of the bottom of the sample storage hole, the spring 3028 applies a flexible force to convert the contact between the sample injection needle 32 and the inner wall of the bottom of the sample storage hole into flexible contact, so that the sample injection needle 32 can be prevented from being damaged.

The X-axis motion assembly 300, the Y-axis motion assembly 301, and the Z-axis motion assembly 302 respectively realize motion in three directions of XYZ, and in this embodiment, a lead screw motor mechanism is adopted. Specifically, the bottom plate 5 is provided with a mounting bracket 303, and the X-axis moving assembly 300 includes an X-axis motor 3000 disposed on the mounting bracket 303, an X-axis screw 3001 drivingly connected to the X-axis motor 3000, and an X-axis slider 3002 (provided with a threaded hole 333 inside) sleeved on the X-axis screw 3001 and in threaded fit with the X-axis screw 3001. The X-axis screw 3001 rotates to move the X-axis slider 3002 screw-engaged therewith along the X-axis. The Y-axis moving component 301 comprises a Y-axis mounting plate 3010 arranged on the X-axis sliding block 3002, a Y-axis motor 3011 arranged on the Y-axis mounting plate 3010, a Y-axis lead 3012 in driving connection with the Y-axis motor 3011, and a Y-axis sliding block 3013 (provided with a threaded hole 333 inside) sleeved on the Y-axis lead 3012 and in threaded fit with the Y-axis lead 3012; the Y-axis screw 3012 is rotated to move the Y-axis slider 3013, which is screw-engaged with it, along the Y-axis. A Z-axis mounting plate 3020 of the Z-axis moving assembly 302 is provided on the Y-axis slider 3013. The Z-axis screw 3022 is rotated to move the Z-axis sliding sleeve 3023 (with the threaded hole 333 therein) threaded therein along the Z-axis.

Further, positioning mechanisms are arranged on the X-axis movement assembly 300, the Y-axis movement assembly 301 and the Z-axis movement assembly 302 and comprise groove-shaped optical couplers and blocking pieces and are used for realizing positioning in all directions. Specifically, an X-axis optical coupler 3003 is arranged on the mounting bracket 303, and an X blocking plate 3004 matched with the X-axis optical coupler 3003 is arranged on the X-axis slider 3002; a Y-axis optical coupler 3014 is arranged on the Y-axis mounting plate 3010, and a Y blocking piece 3015 matched with the Y-axis optical coupler 3014 is arranged on the Y-axis sliding block 3013; a Z-axis optical coupler 3029 is arranged on the Z-axis mounting board 3020, and a Z stop 332 matched with the Z-axis optical coupler 3029 is arranged on the buffer connecting block 33.

In one embodiment, referring to FIGS. 12-13, a detection line segment 223 is connected between the main outlet 222 of the detection chip 22 and the waste line, and the detection line segment 223 is at the detection position of the optical detection assembly 4.

The optical detection assembly 4 includes at least two laser light sources, a transmission light path 44 for transmitting light emitted from the at least two laser light sources to the droplet of the sample to be detected on the detection pipe section 223, at least two lateral detectors for collecting fluorescence generated by the droplet of the sample to be detected on the detection pipe section 223, and a forward detector 49 for collecting laser light after passing through the droplet of the sample to be detected on the detection pipe section 223. The optical detection assembly 4 is mounted on the base plate 5 by a mounting bracket.

In the present embodiment, the laser light source includes 2, a first laser 40 and a second laser 41; the side detector includes 2: a first lateral detector 42 and a second lateral detector 43. The transmission optical path 44 includes a first reflecting mirror 440, a first dichroic mirror 441, and a focusing lens 442, and the optical detection assembly 4 further includes a second dichroic mirror 45, a first optical filter 46, and a second optical filter 47.

The laser emitted by the first laser 40 is reflected by the first reflecting mirror 440 and then transmits the first dichroic mirror 441, the laser emitted by the second laser 41 is reflected by the first dichroic mirror 441, and is converged with the laser of the first laser 40, and is focused onto the detection pipeline section 223 by the focusing lens 442, so as to irradiate the micro-droplets of the sample to be detected; the laser after passing through the micro-droplet of the sample to be detected continues to propagate forward to reach the forward detector 49;

the micro-droplet of the sample to be detected is excited to generate two kinds of fluorescence after being irradiated by two kinds of laser, wherein one kind of fluorescence is reflected by the second dichroic mirror 45 and reaches the first lateral detector 42 after passing through the first optical filter 46; the other is transmitted through a second dichroic mirror 45 and through a second filter 47 to a second lateral detector 43.

Two lasers emit laser with different wavelengths, and the dual fluorescence signal detection is realized by combining 2 lateral detectors, so that the sample utilization rate can be improved, and the significance is more remarkable for precious samples to be detected.

Wherein, the lateral detector collects the fluorescence signal (lateral signal) generated by the excitation of the micro-droplet of the sample to be detected, the forward detector 49 collects the laser signal (forward signal) passing through the micro-droplet of the sample to be detected on the detection pipeline section 223, and the combination of the forward signal and the lateral signal is more beneficial to the data processing in the later period. The forward signal directly characterizes the physical morphology of the droplet, and the fluorescence signal is fluorescence after excitation and does not characterize the morphology of the droplet. And fault judgment is carried out in the subsequent fault maintenance process, and the judgment is conveniently carried out by maintenance personnel by observing the form of the microdroplets in the forward direction. In addition, the fluorescence signal is greatly affected by amplification, and is more uniform if the amplification is good, and is more stray if the amplification is not good. The forward signal is mainly influenced by the physical form of the droplets, and the droplets can be judged quickly by the forward signal to carry out the algorithm processing of fluorescence negative and positive. The method is easy to realize in the peak searching algorithm processing process, and meanwhile, the probability of misreading or leakage is low, and the accuracy is high.

Further, the number of the laser light sources and the lateral detectors can be further expanded on the basis of the above, and more kinds of fluorescence signal detection can be realized.

The working process of the whole instrument is as follows: firstly, storing the micro-droplets of the sample to be detected in the pore plate, then placing the pore plate into a micro-droplet reading instrument, then sucking the sample in the pore plate by using the three-dimensional moving mechanical arm 30, conveying the sample to the detection chip 22 through the liquid conveying system 2, sequentially arranging the micro-droplets of the sample to be detected through the detection pipeline section 223 by virtue of the shearing action of detection oil, and realizing the fluorescence detection of each micro-droplet by using the optical detection component 4.

While the embodiments of the invention have been disclosed above, it is not limited to the applications listed in the description and the embodiments, which are fully applicable in all kinds of fields where the invention is suitable, and further modifications may readily be made by those skilled in the art, and the invention is therefore not limited to the specific details without departing from the general concept defined by the claims and the scope of equivalents.

Claims (10)

1. A droplet reader, comprising: the device comprises a liquid storage assembly, a liquid conveying system, a sample loading assembly and an optical detection assembly;

the liquid storage assembly is used for providing detection oil and collecting waste oil, and the sample loading assembly is used for sucking sample micro-droplets to be detected placed in the micro-droplet reading instrument and conveying the sample micro-droplets to be detected through the liquid conveying system to enable the sample micro-droplets to be detected to be sequentially arranged and pass through the detection position of the optical detection assembly so as to perform fluorescence detection on the sample micro-droplets to be detected.

2. The droplet reader of claim 1, wherein the reservoir assembly comprises a base, a reservoir cartridge slidably disposed on the base, and at least two reservoirs disposed within the reservoir cartridge; the at least two liquid storage tanks at least comprise a detection oil tank and a waste oil tank;

the sample loading assembly comprises a three-dimensional moving mechanical arm, a sample storage device and a sample injection needle arranged on the three-dimensional moving mechanical arm; the sample storage device is used for storing the micro-droplets of the sample to be detected.

3. The droplet reading apparatus according to claim 2, wherein the liquid delivery system comprises an oil pipeline communicated with the detection oil tank, an oil pump disposed on the oil pipeline, a detection chip communicated with an output end of the oil pipeline through an oil inlet, a sample pump, a sample delivery pipeline communicated between the sample pump and a sample inlet of the detection chip, a reversing valve disposed on the sample delivery pipeline, a sample suction pipeline communicated between the reversing valve and a sample inlet needle, a waste oil pipeline communicated between a main outlet of the detection chip and the waste oil tank, and an electromagnetic valve disposed on the waste oil pipeline.

4. A droplet reader according to claim 3 wherein the diverter valve comprises at least a first port, a second port and a third port and is switchable between a first state and a second state, the first state being: the first port is communicated with the second port, and the second port is communicated with the third port; the second state of disconnection is: the second port and the third port are conducted, and the first port and the second port are disconnected;

the second port is communicated with the sample pump through the sample conveying pipeline, the third port is communicated with the sample inlet of the detection chip through the sample conveying pipeline, and the first port is communicated with the sample injection needle through the sample suction pipeline.

5. The droplet reading apparatus according to claim 4, wherein the number of the oil inlets of the detection chip is 2, the detection chip has a cross-shaped micro-channel structure inside, the micro-channel structure forms the sample inlet, 2 oil inlets and a main outlet on four sides of the detection chip, and the 2 oil inlets are symmetrically distributed on two sides of the sample inlet.

6. The droplet reader of claim 2 further comprising a base plate, the reservoir assembly, the fluid delivery system, the loading assembly, and the optical detection assembly being disposed on the base plate;

the liquid storage base is fixedly connected to the bottom plate, and the liquid storage tank comprises a tank body, a tank opening arranged at the top of the tank body, a joint pipe arranged on the tank opening, a joint converter with the lower end inserted in the joint pipe and the upper end extending out, and a tank cover in threaded fit with the joint pipe and used for fixing the lower end of the joint converter in the joint pipe;

a plurality of channels for communicating the inside of the tank body with the outside are arranged in the joint converter;

the channel comprises a horizontal channel and a vertical channel which are communicated with each other, the horizontal channel forms an external pipeline interface on the side surface of the upper end of the joint converter, and the vertical channel forms an internal pipeline interface on the lower end surface of the joint converter.

7. The droplet reading apparatus of claim 6, wherein the lid defines a through hole in a central portion thereof, and an annular flange is disposed on an inner edge of an upper end thereof;

an annular baffle plate is arranged on the periphery of the joint converter, and the outer diameter of the annular baffle plate is smaller than the diameter of the through hole and larger than the inner diameter of the annular flange;

the joint converter is inserted in the joint pipe, and after the tank cover is in threaded connection with the joint pipe, the annular flange is pressed against the upper surface of the annular baffle.

8. The droplet reader of claim 7 wherein the three-dimensional motion robot comprises an X-axis motion assembly disposed on the base plate, a Y-axis motion assembly disposed on the X-axis motion assembly, and a Z-axis motion assembly disposed on the Y-axis motion assembly;

the Z-axis motion assembly comprises a Z-axis mounting plate arranged on the Y-axis motion assembly, a Z-axis motor arranged on the Z-axis mounting plate, a Z-axis screw rod in driving connection with the Z-axis motor and a Z-axis sliding sleeve in threaded fit with the Z-axis screw rod;

the sample injection needle is connected to the Z-axis sliding sleeve through a buffer connecting block.

9. The droplet reading apparatus according to claim 8, wherein the buffer connecting block has a threaded hole, the upper portion of the Z-axis sliding sleeve has a boss, the boss has a through hole, a screw with a diameter smaller than that of the through hole is inserted into the through hole, the lower portion of the screw is fixedly connected to the threaded hole, and the upper end of the screw has a nut with a diameter larger than that of the through hole; a spring is sleeved on the Z-axis sliding sleeve, the upper end of the spring is in contact with the bottom surface of the boss, and the lower end of the spring is in contact with the buffer connecting block;

the buffer connecting block is provided with a via hole with a diameter larger than that of the Z-axis screw rod for the Z-axis screw rod to pass through, and the upper part of the buffer connecting block is also provided with a holding groove for holding the Z-axis sliding sleeve.

10. The droplet reader of claim 5, wherein a detection line segment is connected between the general outlet of the detection chip and the waste line, the detection line segment being at the detection position of the optical detection assembly;

the optical detection assembly comprises at least two laser light sources, a transmission light path for transmitting light emitted by the at least two laser light sources to a sample droplet to be detected on the detection pipeline section, at least two lateral detectors for collecting fluorescence generated by the excitation of the sample droplet to be detected on the detection pipeline section, and a forward detector for collecting laser passing through the sample droplet to be detected on the detection pipeline section.

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