CN104713858B - Full-automatic Time-resolved fluorescence assay device and application method - Google Patents
Full-automatic Time-resolved fluorescence assay device and application method Download PDFInfo
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Abstract
The invention discloses a kind of full-automatic Time-resolved fluorescence assay device and application method, including sample filling unit, time-resolved fluorescence detection unit, concussion hatch unit, microwell plate handling unit, reagent filling unit, some board-washing units, sample-adding platform, reagent rest area, sample rest area, sample needle cleaning unit, left and right rail and control unit；Sample fills unit and microwell plate handling unit is removably mounted within left rail；Reagent filling unit is removably mounted within right guide rail；Board-washing unit includes guide rail, pallet, board-washing head and liquid feeding head, and pallet is arranged on guide rail and moved left and right along guide rail, and board-washing head is arranged on guide rail side by support and moved up and down, and liquid feeding head is arranged on guide rail side by support and moved forward and backward.The present invention realizes scheduling controlling and the optimum organization of different experiments step, can complete test experiments fast and reliablely automatically, improve testing efficiency.
Description
Technical field
The present invention relates to the vitro detection equipment in medicine equipment, a kind of full-automatic Time-resolved fluorescence assay is particularly belonged to
Device.
Background technology
Time resolved fluoro-immunoassay (Time-resolved Fluorescence Immunoassay, referred to as
TRFIA it is) that it is anti-using lanthanide series mark when former is widely used in the important immunoassay method in clinical and scientific research
Former or antibody, according to the luminous characteristics of lanthanide chelate, fluorescence is measured with TIME RESOLVED TECHNIQUE, at the same Detection wavelength and when
Between two parameters carry out signal resolutions, can effectively exclude the interference of non-specific fluorescence, be greatly enhanced the sensitivity of analysis.
Therefore, this method has the features such as low sensitivity height, background values, repeatable measurement.
However, there is presently no the professional automatic equipment for TRFIA Technology designs.Common way is on the market
From general blood treatment platform（The ELISA-STAR of Freedom EVO, HAMILTON companies of such as Tecan companies）Go
Realize the automation of TRFIA experiments.But, because previous platform is not designed exclusively for TRFIA, therefore entered using these platforms
When row TRFIA is tested, some functional modules and actual matching are poor, cause test result inaccurate, and cost is higher.
The content 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 application method,
Rapidly and accurately process can be analyzed by deadline resolved fluorometric, improve detection efficiency, reduce human cost.
In order to solve the above technical problems, the present invention provides a kind of full-automatic Time-resolved fluorescence assay device, including sample
If filling unit, time-resolved fluorescence detection unit, concussion hatch unit, microwell plate handling unit, reagent filling unit, dry-cleaning
Slab element, sample-adding platform, reagent rest area, sample rest area, sample needle cleaning unit, left rail, right guide rail and control unit；
The sample fills unit, can move on left rail and along left rail, for Sample Dilution and to micropore
Sample is filled in plate；
The microwell plate handling unit, can be moved on left rail and along left rail, for by microwell plate in the time
Moved between resolved fluorometric detection unit, concussion hatch unit, board-washing unit and sample-adding platform；
The time-resolved fluorescence detection unit, for carrying out fluoroscopic examination to the sample in microwell plate；
The concussion hatch unit, the concussion frame with some receiving microwell plates, for storing microwell plate and to micropore
Sample in plate carries out concussion incubation；
The reagent fills unit, can move on right guide rail and along right guide rail, thereon equipped with disposable feed head,
For filling reagent into microwell plate；
The board-washing unit, including guide rail, pallet, board-washing head and liquid feeding head, the pallet are arranged on guide rail and along leading
Rail is moved left and right, and the board-washing head is arranged on guide rail side by support and moved up and down along place support, and the liquid feeding head leads to
Support is crossed installed in guide rail side, the liquid feeding head is movable along place support；
The reagent rest area, for depositing reagent, feed head and reagent dilutions cup；
The sample needle cleaning unit, sample rest area, sample-adding platform and reagent rest area are flat located at device from left to right
On platform；
Described control unit, with sample filling unit, time-resolved fluorescence detection unit, concussion hatch unit, microwell plate
Handling unit, reagent filling unit, the pallet in board-washing unit, board-washing head and liquid feeding head are electrically connected with.
Further, the time-resolved fluorescence detection unit and concussion hatch unit are arranged on the lower section of left rail.
Wherein, the sample filling unit has to repeat equipped with one on multiple independent filling heads, each filling head and made
Sampling needle.
It is preferred that, the microwell plate has different identification bar codes, and the microwell plate handling unit has barcode scanner.
It is preferred that, the reagent filling unit has the barcode scanner of identification agent.
Preferably, there is temperature controller and heater in the concussion hatch unit.
The present invention also provides the application method of the 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 it is flat that microwell plate is placed on sample-adding
Moved on platform and by microwell plate handling unit and stored in concussion hatch unit；
Step 2, microwell plate handling unit will be stored in the microwell plate in concussion hatch unit and be moved on sample-adding platform, sample
The sample of sample rest area is filled into the microwell plate on sample-adding platform by this filling unit, often fills a sample, and sample adds
Note unit is moved at sample needle cleaning unit and cleaned once；
Step 3, microwell plate is moved in concussion hatch unit by microwell plate handling unit at sample-adding platform, the micropore
Concussion incubation is carried out in plate earthquake hatch unit；
Step 4, the reagent of reagent filling unit identification agent rest area under the driving of control unit, and configure required try
Agent is inserted in the reagent dilutions cup of reagent rest area；
Step 5, the pallet in board-washing unit is moved along the rail to left end, and microwell plate handling unit will be shaken in hatch unit
Microwell plate take out and be put into pallet, board-washing in board-washing unit head is cleaned several times to microwell plate；
Step 6, microwell plate moves to the right-hand member of guide rail under the drive of pallet, and reagent fills unit by reagent dilutions cup
Reagent be filled into microwell plate；
Step 7, microwell plate moves to the left end of guide rail under the drive of pallet, and microwell plate handling unit will be micro- in pallet
Orifice plate is moved in concussion hatch unit, and concussion incubation is carried out in the microwell plate earthquake hatch unit；
Step 8, the microwell plate shaken in hatch unit is taken out and is put into pallet by microwell plate handling unit, in board-washing unit
Board-washing head microwell plate is cleaned several times；
Step 9, the liquid feeding head in board-washing unit fills enhancing liquid into the micropore of microwell plate, and microwell plate handling unit will be micro-
Orifice plate is moved in concussion hatch unit and carries out concussion incubation；
Step 10, the microwell plate shaken in hatch unit is taken out and is put into time-resolved fluorescence detection by microwell plate handling unit
In unit, fluorescent value is read；
Step 11, microwell plate is taken out temporary earthquake from time-resolved fluorescence detection unit and incubated by microwell plate handling unit
Educate in unit, moved to and be removed on sample-adding platform by microwell plate handling unit again after experiment terminates.
Wherein, in step 5 and step 8, the board-washing head fills cleaning solution into the micropore of microwell plate, again will after immersion
Cleaning solution is siphoned away, and completion is once cleaned.
The structure of each functional module of reasonable Arrangement of the present invention and position, and different experiments step is realized by control unit
Scheduling controlling and optimum organization, can complete test experiments fast and reliablely automatically, improve testing efficiency, reduce artificial
Participate in.
Brief description of the drawings
Fig. 1 is the 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 that sample fills unit；2 be left rail；3 be time-resolved fluorescence detection unit；4 be concussion hatch unit；5 are
Microwell plate handling unit；6 be right guide rail；7 be that reagent fills unit；8 be reagent rest area；9 be board-washing unit；10 is flat for sample-adding
Platform；11 be microwell plate；12 be sample rest area；13 be sample needle cleaning unit；14 be pallet；15 be guide rail；16 be liquid feeding head；
17 be board-washing head.
Embodiment
The present invention is further detailed explanation with embodiment below in conjunction with the accompanying drawings.
The full-automatic Time-resolved fluorescence assay device that the present invention is provided, as shown in figure 1, including sample filling unit 1, when
Between resolved fluorometric detection unit 3, concussion hatch unit 4, microwell plate handling unit 5, reagent filling unit 7, some board-washing units
9th, sample-adding platform 10, reagent rest area 8, sample rest area 12, sample needle cleaning unit 13, left rail 2, right guide rail 6 and control
Unit；
The sample fills unit 1, can move on left rail 2 and along left rail 2, for Sample Dilution and to micro-
Sample is filled in orifice plate 11；
The microwell plate handling unit 5, is robot manipulator structure（With reference to utility model 201320416654.3）, it is arranged on
Can move on left rail 2 and along left rail 2, for by microwell plate 11 in time-resolved fluorescence detection unit 3, concussion hatch unit
4th, moved between board-washing unit 9 and sample-adding platform 10；
The time-resolved fluorescence detection unit 3, for carrying out fluoroscopic examination to the sample in microwell plate 11；
The concussion hatch unit 4, the concussion frame with some receiving microwell plates 11（With reference to utility model
201320416698.6）, for storing microwell plate 11 and carrying out concussion incubation to the sample in microwell plate 11；
The reagent fills unit 7, can move on right guide rail 6 and along right guide rail 6, is sample loading gun structure, thereon
Equipped with disposable feed head（With reference to utility model 201320416034.X）, for filling reagent into microwell plate 11；
The board-washing unit 9, including guide rail 15, pallet 14, board-washing first 17 and liquid feeding first 16, the pallet 14 are arranged on and led
Moved left and right on rail 15 and along guide rail 15, the board-washing first 17 is arranged on the side of guide rail 15 and along above and below the support of place by support
Mobile, the liquid feeding first 16 is arranged on the side of guide rail 15 by support, and the liquid feeding first 16 is movable along place support；
The reagent rest area 8, for depositing reagent, feed head and reagent dilutions cup；
The sample needle cleaning unit 13, sample rest area 12, sample-adding platform 10 and reagent rest area 8 are located at from left to right
On apparatus platform；
Described control unit, with sample filling unit 1, time-resolved fluorescence detection unit 3, concussion hatch unit 4, micropore
Plate handling unit 5, reagent filling unit 7, the pallet 14 in board-washing unit 9, board-washing first 17 and liquid feeding first 16 are electrically connected with.
In said structure, time-resolved fluorescence detection unit 3 is arranged on the lower section of left rail 2 with concussion hatch unit 4,
The space-consuming and volume of device can effectively be reduced.
Sample filling unit 1 has multiple independent filling heads, in the present embodiment, by taking six filling heads as an example, each
One reusable sampling needle is housed, the principle that the sampling needle is replaced using air takes liquid liquid feeding on filling head.More preferably, respectively
Spacing between filling head can be adjusted, and can be met into the microwell plate of different size the need for liquid feeding.
Microwell plate 11 has different identification bar codes, and microwell plate handling unit 5 has barcode scanner.Reagent fills unit
7 have the barcode scanner of identification agent.
In addition, for the need for meeting different tests, there is temperature controller and heater in concussion hatch unit 4.
Further, control unit and a display unit（Display or liquid crystal display）It is connected.
With hepatitis B surface antigen（HBsAg）TRFIA experiment exemplified by, illustrate the application method of aforementioned means.
Step 1, preparation, by sample（Serum or blood plasma）It is placed in sample rest area 12；
Step 2, by the microwell plate 11 of HBsAg projects be positioned over sample-adding platform 10 on, microwell plate handling unit 5 is picked up successively
Microwell plate 11, reads its bar code and is carried in concussion hatch unit 4, according to industrial practice usually using 96 hole micropores
Plate, i.e., can do 96 tests per plate, if any multiple projects, and multiple microwell plates 11 can be stacked on sample-adding platform 10；
Step 3, reagent is positioned over to reagent rest area 8, the reagent of reagent filling unit 7 reading reagent rest area 8, with true
It is required reagent to recognize it；
It is flat that the microwell plate 11 of HBsAg projects is carried to sample-adding by step 4, microwell plate handling unit 5 from concussion hatch unit 4
On platform 10；
Step 5, sample filling unit 1 fill the sample of sample rest area 12 in the microwell plate 11 on sample-adding platform 10
In, per the μ L of hole 100；After often filling once, sample filling unit 1, which is moved at sample needle cleaning unit 13, to be cleaned, to avoid taking
Band pollution；Certainly, sample filling unit 1 can empirically demand percentage dilutes by sample, and the HBsAg projects sample of the present embodiment is not
Need dilution；
Microwell plate 11 after sample-adding is carried to concussion hatch unit by step 6, microwell plate handling unit 5 from sample-adding platform 10
In 4；
Room temperature concussion is incubated 40 minutes in step 7, the earthquake hatch unit 4 of HBsAg microwell plates 11；Because concussion is incubated list
Member 4 has temperature controller and heater, and such as project needs, and can be heated during earthquake incubation；
Step 8, reagent filling unit 7 are by the reagent dilutions cup that each reagent is configured in reagent rest area 8；
Pallet 14 in step 9, board-washing unit 9 moves to the left end of guide rail 15；
Microwell plate 11 is carried to the pallet of board-washing unit 9 by step 10, microwell plate handling unit 5 from concussion hatch unit 4
On 14；Two board-washing units are provided with the present embodiment, and during solving to do the experiment of multiple projects, board-washing element resources are not enough
The problem of；
Step 11, pallet 14 drive microwell plate 11 to move to first 17 position of board-washing, and board-washing unit 9 is to HBsAg microwell plates
11 cleanings 4 times, board-washing first 17 will fill cleaning solution in the micropore on microwell plate 11, soak about one second kind, then cleaning solution is siphoned away,
As once clean；
Pallet 14 in step 12, board-washing unit 9 moves to the right-hand member of guide rail 15 with microwell plate 11；
Step 13, reagent filling unit 7 fill configured good reagent in step 8 in micro- on the pallet of board-washing unit 9
In orifice plate 11；
Pallet 14 in step 14, board-washing unit 9 moves to the left end of guide rail 15 with microwell plate 11, and microwell plate is carried single
Microwell plate 11 is carried in concussion hatch unit 4 by member 5 at pallet 14；
Concussion is incubated 40 minutes in step 15, the earthquake hatch unit 4 of microwell plate 11；
Step 16, repeat step 9,10,11, but board-washing number of times is changed to 6 times；
Step 17, pallet 14 drive microwell plate 11 to be moved to first 16 position of liquid feeding, and liquid feeding first 16 is toward in microwell plate 11
Filling enhancing liquid（Special agent in time resolution kit）, per the μ L of hole 200；
Pallet 14 in step 18, board-washing unit 9 moves to the left end of guide rail 15 with microwell plate 11, and microwell plate is carried single
Microwell plate 11 is carried in concussion hatch unit 4 by member 5；
Concussion is incubated 5 minutes in step 19, the earthquake hatch unit 4 of microwell plate 11；
Microwell plate 11 is taken out and is carried to time resolution by step 20, microwell plate handling unit 5 from concussion hatch unit 4
In fluorescence detection unit 3, fluorescent value is read；
Step 21, microwell plate handling unit 5 take out microwell plate 11 from time-resolved fluorescence detection unit 3, and carry
Kept in concussion hatch unit 4；
Step 22, experiment terminate, and microwell plate 11 is carried to sample-adding platform 10 by microwell plate handling unit 5, and by operator
Collect away.
It is above-mentioned only with hepatitis B surface antigen（HBsAg）TRFIA experiments exemplified by illustrate, when carrying out other TRFIA experiment, wash
Plate number of times, concussion incubation time and temperature, reagent and the enhancing type such as liquid can be converted accordingly.
The structure of each functional module of reasonable Arrangement of the present invention and position, and different experiments step is realized by control unit
Scheduling controlling and optimum organization, can complete test experiments fast and reliablely automatically, improve testing efficiency, reduce artificial
Participate in.
The present invention is described in detail above by specific embodiment, the embodiment is only the preferable of the present invention
Embodiment, it not limits the invention.Without departing from the principles of the present invention, those skilled in the art is to each
Equivalent replacement and improvement that structure of unit etc. is made, are regarded as in the technology category of the invention protected.
Claims (7)
1. a kind of full-automatic Time-resolved fluorescence assay device, it is characterised in that including sample filling unit (1), time resolution
Fluorescence detection unit (3), concussion hatch unit (4), microwell plate handling unit (5), reagent filling unit (7), some board-washing lists
First (9), sample-adding platform (10), reagent rest area (8), sample rest area (12), sample needle cleaning unit (13), left rail (2),
Right guide rail (6) and control unit；
Sample filling unit (1), can be moved on left rail (2) and along left rail (2), for Sample Dilution and to
Filling sample in microwell plate (11)；
The microwell plate handling unit (5), on left rail (2) and can be mobile along left rail (2), for by microwell plate
(11) moved between time-resolved fluorescence detection unit (3), concussion hatch unit (4), board-washing unit (9) and sample-adding platform (10)
It is dynamic；
The time-resolved fluorescence detection unit (3), for carrying out fluoroscopic examination to the sample in microwell plate (11)；
It is described concussion hatch unit (4), with it is some receiving microwell plates (11) concussion framves, for store microwell plate (11) and
Concussion incubation is carried out to the sample in microwell plate (11)；
Reagent filling unit (7), on right guide rail (6) and can be mobile along right guide rail (6), thereon equipped with disposably adding
Sample head, for the filling reagent into microwell plate (11)；
The board-washing unit (9), including guide rail (15), pallet (14), board-washing head (17) and liquid feeding head (16), the pallet (14)
Moved left and right on guide rail (15) and along guide rail (15), the board-washing head (17) is arranged on guide rail (15) side by support
And moved up and down along place support, the liquid feeding head (16) is arranged on guide rail (15) side, liquid feeding head (16) edge by support
Place support is moved forward and backward；
The reagent rest area (8), for depositing reagent, feed head and reagent dilutions cup；
The sample needle cleaning unit (13), sample rest area (12), sample-adding platform (10) and reagent rest area (8) are from left to right
On apparatus platform；
Described control unit, with sample filling unit (1), time-resolved fluorescence detection unit (3), concussion hatch unit (4), micro-
Orifice plate handling unit (5), reagent filling unit (7), the pallet (14) in board-washing unit (9), board-washing head (17) and liquid feeding head
(16) it is electrically connected with；
Wherein, the time-resolved fluorescence detection unit (3) and concussion hatch unit (4) are arranged on the lower section of left rail (2).
2. full-automatic Time-resolved fluorescence assay device according to claim 1, it is characterised in that the sample filling is single
First (1), which has, is equipped with a reusable sampling needle on multiple independent filling heads, each filling head.
3. full-automatic Time-resolved fluorescence assay device according to claim 1, it is characterised in that the microwell plate (11)
With different identification bar codes, the microwell plate handling unit (5) has barcode scanner.
4. full-automatic Time-resolved fluorescence assay device according to claim 1, it is characterised in that the reagent filling is single
First (7) have the barcode scanner of identification agent.
5. full-automatic Time-resolved fluorescence assay device according to claim 1, it is characterised in that the concussion is incubated single
There is temperature controller and heater in first (4).
6. the application method of Time-resolved fluorescence assay device as claimed in claim 1 full-automatic, it is characterised in that including with
Lower step：
Step 1, sample is placed into sample rest area (12), and reagent is placed into reagent rest area (8), and microwell plate (11) is placed on
On sample-adding platform (10) and moved in concussion hatch unit (4) and stored by microwell plate handling unit (5)；
Step 2, it is flat that the microwell plate (11) that microwell plate handling unit (5) will be stored in concussion hatch unit (4) is moved to sample-adding
On platform (10), the sample of sample rest area (12) is filled into the microwell plate on sample-adding platform (10) by sample filling unit (1)
(11) in, a sample is often filled, sample filling unit (1) is moved to the cleaning of sample needle cleaning unit (13) place once；
Step 3, microwell plate (11) is moved to concussion hatch unit (4) by microwell plate handling unit (5) from sample-adding platform (10)
In, concussion incubation is carried out in microwell plate (11) the earthquake hatch unit (4)；
Step 4, reagent filling unit (7) under the driving of control unit identification agent rest area (8) reagent, and configure needed for
Reagent is inserted in the reagent dilutions cup of reagent rest area (8)；
Step 5, the pallet (14) in board-washing unit (9) moves to left end along guide rail (15), and microwell plate handling unit (5) will shake
Microwell plate (11) in hatch unit (4), which takes out, to be put into pallet (14), and the board-washing head (17) in board-washing unit (9) is to microwell plate
(11) cleaned several times；
Step 6, microwell plate (11) moves to the right-hand member of guide rail (15) under the drive of pallet (14), and reagent filling unit (7) will
Reagent in reagent dilutions cup is filled into microwell plate (11)；
Step 7, microwell plate (11) moves to the left end of guide rail (15), microwell plate handling unit (5) under the drive of pallet (14)
Microwell plate (11) in pallet (14) is moved in concussion hatch unit (4), microwell plate (11) the earthquake hatch unit
(4) concussion incubation is carried out in；
Step 8, the microwell plate (11) shaken in hatch unit (4) is taken out and is put into pallet (14) by microwell plate handling unit (5),
Board-washing head (17) in board-washing unit (9) is cleaned several times to microwell plate (11)；
Step 9, the liquid feeding head (16) in board-washing unit (9) fills enhancing liquid into the micropore of microwell plate (11), and microwell plate is carried
Microwell plate (11) is moved in concussion hatch unit (4) and carries out concussion incubation by unit (5)；
Step 10, microwell plate handling unit (5) takes out the microwell plate (11) shaken in hatch unit (4) that to be put into time resolution glimmering
In optical detecting unit (3), fluorescent value is read；
Step 11, microwell plate (11) is taken out and is temporarily stored in by microwell plate handling unit (5) from time-resolved fluorescence detection unit (3)
Shake in hatch unit (4), moved to and be removed on sample-adding platform (10) by microwell plate handling unit (5) again after experiment terminates.
7. the application method of full-automatic Time-resolved fluorescence assay device according to claim 6, it is characterised in that in step
Rapid 5 and step 8 in, board-washing head (17) fills cleaning solution into the micropore of microwell plate (11), again inhales cleaning solution after immersion
Walk, completion is once cleaned.
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Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105754842B (en) * | 2016-04-29 | 2018-08-28 | 珠海迪尔生物工程有限公司 | A kind of cold chain storage extraction auxiliary reagent sample adding device |
| 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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