CN104713858A - Automatic time-resolved fluorescence assay device and application method thereof - Google Patents

Automatic time-resolved fluorescence assay device and application method thereof Download PDF

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Publication number
CN104713858A
CN104713858A CN201310674089.5A CN201310674089A CN104713858A CN 104713858 A CN104713858 A CN 104713858A CN 201310674089 A CN201310674089 A CN 201310674089A CN 104713858 A CN104713858 A CN 104713858A
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unit
microwell plate
sample
reagent
concussion
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CN201310674089.5A
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CN104713858B (en
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许海峰
许明
樊振昌
同绪刚
周德
邓永锦
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Suzhou Sym Bio Lifescience Co Ltd
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Suzhou Sym Bio Lifescience Co Ltd
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Abstract

The invention discloses an automatic time-resolved fluorescence assay device and an application method thereof. The device comprises a sample injection unit, a time-resolved fluorescence assay unit, a vibration incubation unit, a micropore plate delivering unit, a reagent injecting unit, a plurality of washing plate units, a sample injecting platform, a reagent placing area, a sample placing area, an injecting needle washing unit, a left guide rail, a right guide rail, and a control unit. The sample injecting unit and the micropore plate delivering unit are movably installed on the left guide rail; the reagent injecting unit is movably arranged on the right guide rail, each washing plate unit comprises a guide rail, a tray, a washing plate head, and a liquid feeding head, the tray is arranged on the guide rail and moves horizontally along the guide rail, the washing plate head is arranged on one side of the guide rail and moves vertically, and the liquid feeding head is installed on one side of the guide rail through a bracket and moves horizontally. The provided device can modulate and optimize different experiment steps, is capable of finishing testing experiments rapidly and automatically, and thus improves the testing efficiency.

Description

Full-automatic Time-resolved fluorescence assay device and using method
Technical field
The present invention relates to the vitro detection equipment in medicine equipment, specifically belong to a kind of full-automatic Time-resolved fluorescence assay device.
Background technology
Time resolved fluoro-immunoassay (Time-resolved Fluorescence Immunoassay, be called for short TRFIA) be current a kind of important immune analysis method be widely used in clinical and scientific research, it utilizes lanthanide series labelled antigen or antibody, according to the luminous characteristics of lanthanide chelate, fluorescence is measured by TIME RESOLVED TECHNIQUE, determined wavelength and time two parameters carry out signal resolution simultaneously, effectively can get rid of the interference of non-specific fluorescence, greatly improve the sensitivity of analysis.Therefore, the method have highly sensitive, background values is low, can the feature such as duplicate measurements.
But, at present also not for the professional automatic equipment of TRFIA technical design.Way common is on the market the robotization of selecting general blood processing platform (ELISA-STAR etc. as Freedom EVO, HAMILTON company of Tecan company) to go to realize TRFIA experiment.But, due to previous platform and non-expert design for TRFIA, when therefore utilizing these platforms to carry out TRFIA experiment, some functional module is poor with actual matching, cause test result inaccurate, and cost is higher.
Summary of the invention
The technical problem to be solved in the present invention is to provide a kind of full-automatic Time-resolved fluorescence assay device and using method, can deadline resolved fluorometric analytic process rapidly and accurately, improves detection efficiency, reduces human cost.
For solving the problems of the technologies described above, the invention provides a kind of full-automatic Time-resolved fluorescence assay device, comprise sample filling unit, unit is hatched in time-resolved fluorescence detecting unit, concussion, microwell plate handling unit, reagent filling unit, somely wash Slab element, application of sample platform, reagent rest area, sample rest area, sample needle cleaning unit, left rail, right guide rail and control module;
Described sample filling unit, is arranged on left rail and also can moves along left rail, for Sample Dilution and sample of annotating in microwell plate;
Described microwell plate handling unit, is arranged on left rail and also can moves along left rail, for microwell plate is hatched unit in time-resolved fluorescence detecting unit, concussion, washed between Slab element and application of sample platform and move;
Described time-resolved fluorescence detecting unit, for carrying out fluoroscopic examination to the sample in microwell plate;
Unit is hatched in described concussion, has the concussion frame of some accommodation microwell plates, hatches for storing microwell plate and carrying out concussion to the sample in microwell plate;
Described reagent filling unit, is arranged on right guide rail and also can moves along right guide rail, it is equipped with disposable feed head, for reagent of annotating in microwell plate;
Describedly wash Slab element, comprise guide rail, pallet, wash wrench and liquid feeding head, described pallet to be arranged on guide rail and to move left and right along guide rail, described wrench of washing to be moved up and down along place support in guide rail side by support installing, described liquid feeding head is by support installing in guide rail side, and this liquid feeding head moves forward and backward along place support;
Described reagent rest area, for depositing reagent, feed head and reagent dilutions cup;
Described sample needle cleaning unit, sample rest area, application of sample platform and reagent rest area are located on apparatus platform from left to right;
Described control module, with sample annotate unit, time-resolved fluorescence detecting unit, shake hatch unit, microwell plate handling unit, reagent annotate unit, the pallet washed in Slab element, wash wrench and liquid feeding head is electrically connected.
Further, the below of cellular installation at left rail is hatched in described time-resolved fluorescence detecting unit and concussion.
Wherein, described sample filling unit has multiple independently filling head, each filling head is equipped with a reusable sampling needle.
Preferably, described microwell plate has different identification bar codes, and described microwell plate handling unit has barcode scanner.
Preferably, described reagent filling unit has the barcode scanner of identification agent.
Preferably, described concussion is hatched in unit and is had temperature controller and well heater.
The present invention also provides the using method of described full-automatic Time-resolved fluorescence assay device, comprises the following steps:
Step 1, sample is placed into sample rest area, and reagent is placed into reagent rest area, and microwell plate to be placed on application of sample platform and to move to concussion by microwell plate handling unit and hatches in unit and store;
Step 2, microwell plate handling unit moves on application of sample platform by being stored in the concussion microwell plate of hatching in unit, the sample of sample rest area is filled in the microwell plate on application of sample platform by sample filling unit, an often filling sample, sample filling unit moves to the cleaning of sample needle cleaning unit place once;
Step 3, microwell plate is moved to concussion from application of sample platform and hatches in unit by microwell plate handling unit, and described microwell plate earthquake is hatched in unit and carried out concussion and hatch;
Step 4, the reagent of reagent filling unit identification agent rest area under the driving of control module, and the required reagent of configuration is inserted in the reagent dilutions cup of reagent rest area;
Step 5, washes pallet in Slab element along guide rail movement to left end, and by concussion, the microwell plate of hatching in unit takes out and puts into pallet microwell plate handling unit, and the wrench of washing washed in Slab element carries out several times cleaning to microwell plate;
Step 6, microwell plate moves to the right-hand member of guide rail under the drive of pallet, and the reagent in reagent dilutions cup is filled in microwell plate by reagent filling unit;
Step 7, microwell plate moves to the left end of guide rail under the drive of pallet, and the microwell plate in pallet is moved to concussion and hatches in unit by microwell plate handling unit, and described microwell plate earthquake is hatched in unit and carried out concussion and hatch;
Step 8, pallet is put in the microwell plate taking-up that concussion is hatched in unit by microwell plate handling unit, and the wrench of washing washed in Slab element carries out several times cleaning to microwell plate;
Step 9, the liquid feeding head washed in Slab element is annotated and is strengthened liquid in the micropore of microwell plate, and microwell plate is moved to concussion and hatches in unit and carry out concussion and hatch by microwell plate handling unit;
Step 10, time-resolved fluorescence detecting unit is put in the microwell plate taking-up that concussion is hatched in unit by microwell plate handling unit, reads fluorescent value;
Step 11, microwell plate takes out and is temporarily stored in concussion and hatches in unit by microwell plate handling unit from time-resolved fluorescence detecting unit, moves on application of sample platform removed again after experiment terminates by microwell plate handling unit.
Wherein, in step 5 and step 8, described in wash wrench fill cleansing solution in the micropore of microwell plate, again cleansing solution is siphoned away after immersion, completes and once clean.
The structure of each functional module of reasonable Arrangement of the present invention and position, and the scheduling controlling and the optimal combination that are realized different experiments step by control module, automatically can be completed test experiments to fast and reliable, improve testing efficiency, decrease artificial participation.
Accompanying drawing explanation
Fig. 1 is structural representation of the present invention;
Fig. 2 is the structural representation of cleaning unit in the present invention.
Wherein description of reference numerals is as follows:
1 is sample filling unit; 2 is left rail; 3 is time-resolved fluorescence detecting unit; 4 hatch unit for concussion; 5 is microwell plate handling unit; 6 is right guide rail; 7 is reagent filling unit; 8 is reagent rest area; 9 for washing Slab element; 10 is application of sample platform; 11 is microwell plate; 12 is sample rest area; 13 is sample needle cleaning unit; 14 is pallet; 15 is guide rail; 16 is liquid feeding head; 17 for washing wrench.
Embodiment
Below in conjunction with accompanying drawing and embodiment, the present invention is further detailed explanation.
Full-automatic Time-resolved fluorescence assay device provided by the invention, as shown in Figure 1, comprise sample filling unit 1, time-resolved fluorescence detecting unit 3, concussion hatch unit 4, microwell plate handling unit 5, reagent filling unit 7, somely wash Slab element 9, application of sample platform 10, reagent rest area 8, sample rest area 12, sample needle cleaning unit 13, left rail 2, right guide rail 6 and control module;
Described sample filling unit 1, is arranged on left rail 2 and also can moves along left rail 2, for Sample Dilution and sample of annotating in microwell plate 11;
Described microwell plate handling unit 5, for robot manipulator structure (with reference to utility model 201320416654.3), it is arranged on left rail 2 and also can moves along left rail 2, for microwell plate 11 is hatched unit 4 in time-resolved fluorescence detecting unit 3, concussion, washed between Slab element 9 and application of sample platform 10 and move;
Described time-resolved fluorescence detecting unit 3, for carrying out fluoroscopic examination to the sample in microwell plate 11;
Unit 4 is hatched in described concussion, has the concussion frame (with reference to utility model 201320416698.6) of some accommodation microwell plates 11, hatches for storing microwell plate 11 and carrying out concussion to the sample in microwell plate 11;
Described reagent filling unit 7, being arranged on right guide rail 6 and also can moving along right guide rail 6, is sample loading gun structure, it is equipped with disposable feed head (with reference to utility model 201320416034.X), for reagent of annotating in microwell plate 11;
Describedly wash Slab element 9, comprise guide rail 15, pallet 14, wash wrench 17 and liquid feeding head 16, described pallet 14 to be arranged on guide rail 15 and to move left and right along guide rail 15, described wrench 17 of washing to be moved up and down along place support in guide rail 15 side by support installing, described liquid feeding head 16 is by support installing in guide rail 15 side, and this liquid feeding head 16 moves forward and backward along place support;
Described reagent rest area 8, for depositing reagent, feed head and reagent dilutions cup;
Described sample needle cleaning unit 13, sample rest area 12, application of sample platform 10 and reagent rest area 8 are located on apparatus platform from left to right;
Described control module, with sample annotate unit 1, time-resolved fluorescence detecting unit 3, shake hatch unit 4, microwell plate handling unit 5, reagent annotate unit 7, the pallet 14 washed in Slab element 9, wash wrench 17 and liquid feeding head 16 is electrically connected.
In said structure, the below that unit 4 is arranged on left rail 2 is hatched in time-resolved fluorescence detecting unit 3 and concussion, effectively can reduce taking up room and volume of device.
Sample filling unit 1 has multiple independently filling head, and in the present embodiment, for six filling heads, each filling head is equipped with a reusable sampling needle, this sampling needle utilizes the principle of air displacement to get liquid liquid feeding.Better, the spacing between each filling head can regulate, and can meet the needs of liquid feeding in the microwell plate of different size.
Microwell plate 11 has different identification bar codes, and microwell plate handling unit 5 has barcode scanner.Reagent filling unit 7 has the barcode scanner of identification agent.
In addition, be the needs of satisfied difference test, concussion is hatched in unit 4 has temperature controller and well heater.
Further, control module is connected with a display unit (display or liquid crystal display).
Test for the TRFIA of hepatitis B surface antigen (HBsAg), the using method of aforementioned means is described.
Step 1, preliminary work, be placed in sample rest area 12 by sample (serum or blood plasma);
Step 2, the microwell plate 11 of HBsAg project is positioned on application of sample platform 10, microwell plate handling unit 5 picks up microwell plate 11 successively, reading its bar code and being carried to concussion hatches in unit 4, usually 96 hole microwell plates are used according to industrial practice, namely every plate can do 96 tests, if any multiple project, multiple microwell plate 11 can be stacked on application of sample platform 10;
Step 3, reagent is positioned over reagent rest area 8, the reagent of reagent filling unit 7 reading reagent rest area 8, to confirm that it is required reagent;
The microwell plate 11 of HBsAg project is hatched unit 4 from concussion and is carried to application of sample platform 10 by step 4, microwell plate handling unit 5;
In the microwell plate 11 that the sample of sample rest area 12 is annotated on application of sample platform 10 by step 5, sample filling unit 1, every hole 100 μ L; Often annotate once, sample filling unit 1 all moves to the cleaning of sample needle cleaning unit 13 place, to avoid Residual contamination; Certainly, sample filling unit 1 can by sample empirically demand percentage dilution, and the HBsAg project sample of the present embodiment does not need dilution;
Microwell plate 11 after application of sample is carried to concussion from application of sample platform 10 and hatches unit 4 by step 6, microwell plate handling unit 5;
Step 7, HBsAg microwell plate 11 earthquake are hatched room temperature concussion in unit 4 and are hatched 40 minutes; Because concussion is hatched unit 4 and had temperature controller and well heater, as project needs, can heat in the earthquake process of hatching;
Step 8, reagent filling unit 7 is by the reagent dilutions cup be configured at by each reagent in reagent rest area 8;
Step 9, the pallet 14 washed in Slab element 9 move to the left end of guide rail 15;
Microwell plate 11 is hatched unit 4 from concussion and is carried to and washes the pallet 14 of Slab element 9 by step 10, microwell plate handling unit 5; Be provided with two in the present embodiment and wash Slab element, during to solve the experiment doing multiple project, wash the problem of Slab element inadequate resource;
Step 11, pallet 14 drive microwell plate 11 to move to and wash wrench 17 position, wash Slab element 9 pairs of HBsAg microwell plates 11 and clean 4 times, wash wrench 17 and fill cleansing solution by the micropore on microwell plate 11, soak about one second kind, again cleansing solution is siphoned away, be and once clean;
Step 12, the pallet 14 washed in Slab element 9 move to the right-hand member of guide rail 15 with microwell plate 11;
The reagent configured in step 8 is annotated in washing in the microwell plate 11 on Slab element 9 pallet by step 13, reagent filling unit 7;
Step 14, the pallet 14 washed in Slab element 9 move to the left end of guide rail 15 with microwell plate 11, microwell plate 11 is carried to concussion from pallet 14 and hatches in unit 4 by microwell plate handling unit 5;
Step 15, microwell plate 11 earthquake are hatched concussion in unit 4 and are hatched 40 minutes;
Step 16, repeat step 9,10,11, but wash plate number of times and change 6 times into;
Step 17, pallet 14 drive microwell plate 11 to move to liquid feeding head 16 position, and liquid feeding head 16 is annotated and strengthened liquid (special agent in time resolution kit), every hole 200 μ L in microwell plate 11;
Step 18, the pallet 14 washed on Slab element 9 move to the left end of guide rail 15 with microwell plate 11, microwell plate 11 is carried to concussion and hatches in unit 4 by microwell plate handling unit 5;
Step 19, microwell plate 11 earthquake are hatched concussion in unit 4 and are hatched 5 minutes;
Microwell plate 11 is hatched unit 4 from concussion and is taken out and be carried in time-resolved fluorescence detecting unit 3 by step 20, microwell plate handling unit 5, reads fluorescent value;
Microwell plate 11 takes out by step 21, microwell plate handling unit 5 from time-resolved fluorescence detecting unit 3, and is carried to concussion and hatches unit 4 and keep in;
Step 22, experiment terminate, and microwell plate 11 is carried to application of sample platform 10 by microwell plate handling unit 5, and is collected away by operator.
Above-mentioned only experiment for the TRFIA of hepatitis B surface antigen (HBsAg) illustrates, when carrying out other TRFIA experiment, washes plate number of times, type such as concussion incubation time and temperature, reagent and enhancing liquid etc. can correspondingly convert.
The structure of each functional module of reasonable Arrangement of the present invention and position, and the scheduling controlling and the optimal combination that are realized different experiments step by control module, automatically can be completed test experiments to fast and reliable, improve testing efficiency, decrease artificial participation.
Above by specific embodiment to invention has been detailed description, this embodiment is only preferred embodiment of the present invention, and it not limits the invention.Without departing from the principles of the present invention, the equivalent replacement that the structure etc. of those skilled in the art to each unit is made and improvement, all should be considered as in the technology category protected in the present invention.

Claims (8)

1. a full-automatic Time-resolved fluorescence assay device, it is characterized in that, comprise sample filling unit (1), time-resolved fluorescence detecting unit (3), concussion hatch unit (4), microwell plate handling unit (5), reagent filling unit (7), somely wash Slab element (9), application of sample platform (10), reagent rest area (8), sample rest area (12), sample needle cleaning unit (13), left rail (2), right guide rail (6) and control module;
Described sample filling unit (1), being arranged on left rail (2) above also can be mobile along left rail (2), for Sample Dilution with to microwell plate (11) middle filling sample;
Described microwell plate handling unit (5), being arranged on left rail (2) above also can be mobile along left rail (2), for microwell plate (11) is hatched unit (4) in time-resolved fluorescence detecting unit (3), concussion, washed between Slab element (9) and application of sample platform (10) and move;
Described time-resolved fluorescence detecting unit (3), for carrying out fluoroscopic examination to the sample in microwell plate (11);
Unit (4) is hatched in described concussion, has the concussion frame of some accommodation microwell plates (11), hatches for storing microwell plate (11) and carrying out concussion to the sample in microwell plate (11);
Described reagent filling unit (7), being arranged on right guide rail (6) above also can be mobile along right guide rail (6), it is equipped with disposable feed head, for filling reagent in microwell plate (11);
Describedly wash Slab element (9), comprise guide rail (15), pallet (14), wash wrench (17) and liquid feeding head (16), described pallet (14) is arranged on guide rail (15) and goes up and move left and right along guide rail (15), described wrench (17) of washing is moved up and down along place support in guide rail (15) side by support installing, described liquid feeding head (16) is by support installing in guide rail (15) side, and this liquid feeding head (16) moves forward and backward along place support;
Described reagent rest area (8), for depositing reagent, feed head and reagent dilutions cup;
Described sample needle cleaning unit (13), sample rest area (12), application of sample platform (10) and reagent rest area (8) are located on apparatus platform from left to right;
Described control module, with sample annotate unit (1), time-resolved fluorescence detecting unit (3), shake hatch unit (4), microwell plate handling unit (5), reagent annotate unit (7), the pallet (14) washed in Slab element (9), wash wrench (17) and liquid feeding head (16) is electrically connected.
2. full-automatic Time-resolved fluorescence assay device according to claim 1, is characterized in that, the below that unit (4) is arranged on left rail (2) is hatched in described time-resolved fluorescence detecting unit (3) and concussion.
3. full-automatic Time-resolved fluorescence assay device according to claim 1, is characterized in that, described sample filling unit (1) has multiple independently filling head, each filling head is equipped with a reusable sampling needle.
4. full-automatic Time-resolved fluorescence assay device according to claim 1, is characterized in that, described microwell plate (11) has different identification bar codes, and described microwell plate handling unit (5) has barcode scanner.
5. full-automatic Time-resolved fluorescence assay device according to claim 1, is characterized in that, described reagent filling unit (7) has the barcode scanner of identification agent.
6. full-automatic Time-resolved fluorescence assay device according to claim 1, is characterized in that, described concussion is hatched in unit (4) has temperature controller and well heater.
7. the using method of full-automatic Time-resolved fluorescence assay device as claimed in claim 1, is characterized in that, comprise the following steps:
Step 1, sample is placed into sample rest area (12), and reagent is placed into reagent rest area (8), and it is upper and move to concussion by microwell plate handling unit (5) and hatch in unit (4) and store that microwell plate (11) is placed on application of sample platform (10);
Step 2, microwell plate handling unit (5) moves on application of sample platform (10) by being stored in the concussion microwell plate (11) of hatching in unit (4), the sample of sample rest area (12) is filled in the microwell plate (11) on application of sample platform (10) by sample filling unit (1), an often filling sample, sample filling unit (1) moves to the cleaning of sample needle cleaning unit (13) place once;
Step 3, microwell plate (11) is moved to concussion from application of sample platform (10) and hatches in unit (4) by microwell plate handling unit (5), and described microwell plate (11) earthquake is hatched and carried out concussion in unit (4) and hatch;
Step 4, reagent filling unit (7) reagent of identification agent rest area (8) under the driving of control module, and the required reagent of configuration is inserted in the reagent dilutions cup of reagent rest area (8);
Step 5, the pallet (14) washed in Slab element (9) moves to left end along guide rail (15), microwell plate handling unit (5) microwell plate (11) of concussion being hatched in unit (4) takes out puts into pallet (14), and the wrench (17) of washing washed in Slab element (9) carries out several times cleaning to microwell plate (11);
Step 6, microwell plate (11) moves to the right-hand member of guide rail (15) under the drive of pallet (14), and the reagent in reagent dilutions cup is filled in microwell plate (11) by reagent filling unit (7);
Step 7, microwell plate (11) moves to the left end of guide rail (15) under the drive of pallet (14), microwell plate (11) in pallet (14) is moved to concussion and hatches in unit (4) by microwell plate handling unit (5), and described microwell plate (11) earthquake is hatched and carried out concussion in unit (4) and hatch;
Step 8, microwell plate handling unit (5) microwell plate (11) of concussion being hatched in unit (4) takes out puts into pallet (14), and the wrench (17) of washing washed in Slab element (9) carries out several times cleaning to microwell plate (11);
Step 9, the liquid feeding head (16) washed in Slab element (9) is annotated and is strengthened liquid in the micropore of microwell plate (11), and microwell plate (11) is moved to concussion and hatches and carry out concussion in unit (4) and hatch by microwell plate handling unit (5);
Step 10, microwell plate handling unit (5) microwell plate (11) of concussion being hatched in unit (4) takes out puts into time-resolved fluorescence detecting unit (3), reads fluorescent value;
Step 11, microwell plate (11) takes out and is temporarily stored in concussion and hatches in unit (4) by microwell plate handling unit (5) from time-resolved fluorescence detecting unit (3), moves to application of sample platform (10) upper removed after experiment terminates by microwell plate handling unit (5).
8. the using method of full-automatic Time-resolved fluorescence assay device according to claim 7, it is characterized in that, in step 5 and step 8, described in wash wrench (17) fill cleansing solution in the micropore of microwell plate (11), again cleansing solution is siphoned away after immersion, complete and once clean.
CN201310674089.5A 2013-12-11 2013-12-11 Full-automatic Time-resolved fluorescence assay device and application method Active CN104713858B (en)

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CN105891180A (en) * 2016-06-14 2016-08-24 珀金埃尔默医学诊断产品(上海)有限公司 Novel full-automatic time resolution fluorescence analysis device and use method thereof
CN113156101A (en) * 2021-04-08 2021-07-23 中国科学院苏州生物医学工程技术研究所 Multi-wavelength time-resolved fluorescence immunoassay analyzer
CN114152496A (en) * 2021-12-31 2022-03-08 河南赛诺特生物技术有限公司 Automatic immunohistochemical dyeing machine
CN115254738A (en) * 2022-07-26 2022-11-01 深圳市爱康生物科技股份有限公司 Plate washing mechanism and in-vitro detection device

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