CN103513024B - Full-automatic immunochemical instrument and using method thereof - Google Patents
Full-automatic immunochemical instrument and using method thereof Download PDFInfo
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- CN103513024B CN103513024B CN201210206355.7A CN201210206355A CN103513024B CN 103513024 B CN103513024 B CN 103513024B CN 201210206355 A CN201210206355 A CN 201210206355A CN 103513024 B CN103513024 B CN 103513024B
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Abstract
The present invention relates to life science and medical research field, disclose a kind of Full-automatic immunochemical instrument and using method thereof.In the present invention, Full-automatic immunochemical instrument can realize overall process automatic liquid-feeding and sucking-off waste liquid, the mode in ware hole is not changed by changing liquid, frequent moving tissue section sample and damage is caused to it or causes displacement in operation can be avoided, and, experimenter also can be avoided directly to contact noxious material and damage health.By instrument automatic operation, reduce labor intensity, raise the efficiency, can human error be exempted, realize robotization and standardization.Liquid distribution and liquid discharge structure and fluid pipeline can move up and down, and reduce when liquid feeding and imbibition, raising, making more convenient to operate when changing reaction utensil.Liquid distribution and liquid discharge structure and fluid pipeline can all around move, and can realize batch processing to reaction utensil hole matrix, efficiency is higher.
Description
Technical field
The present invention relates to life science and medical research field, the full-automatic immunocytochemical technique of the particularly full-automatic immunohistochemistry technology of histopathologic slide, and cultured cell.
Background technology
Immunochemistry dyeing comprises immunohistochemistry and immunocytochemistry, it is classical, conventional and that requirement is very rigorous laboratory work, immunochemistry is very responsive for the albumen change of tissue, cell, can locate, again can sxemiquantitative, is the very valuable detection technique means of one.Expression, with regard to the existence form of histotomy, mainly contains two kinds of modes: paster method and floating (drift) sheet method.Floating sheet method refers to that histotomy is floating, free and carries out in a liquid hatching, reacting, and waits the process of hatching to terminate and after reacting quality, then is mounted on glass sheet by histotomy.The benefit of this method is exactly for comparatively slab (can observe body comparatively maxicell overall picture or structure), can incubation reaction obtain comparatively thorough, fully, completely, true, be that paster method can not be compared.
At present, paster method achieves robotization, but floating sheet method and the immunocytochemical technique with double dish or culture plate cultured cell, be still manual operation.Artificial behaviour is wasted time and energy, operate miss, experiment parameter change etc., easily cause experimental result unstable.
The present inventor finds, problem is in the following areas deposited in manual operations:
1, time-consumingly to take a lot of work: immunochemical procedures process at least needs 7 to 8 hours consuming time each time, and even more than 1 to 2 days, step was many, dry as dust; Each experiment can only process finite sample, as 5 to 7 examples;
2, easily make mistakes: the antibody that immunochemical procedures is used each time and reagent type reach tens kinds, manual operations easily mistake adds or forgets to add, and the reaction time can not accurately control to hold with standard;
3, between different experiments batch, due to multifactorial impact, be difficult to realize reappearing completely and matching, differences between batches are difficult to prevent.
4, tissue section sample easily damages: in manual process, needs frequent moving tissue section sample, easily causes damage to it, and easily causes sample displacement with dragging for into pulling action out;
5, damage healthy: the antibody that immunochemical procedures is used and reagent have severe toxicity a bit, and if diaminobenzidine (DAB) is clear and definite strong carcinogen, in manual process, experimenter directly contacts harmful liquid, damages healthy.
Therefore, need now one badly and can realize overall process automatic operation, both can not damage, displacement tissue section sample, also can not damage healthy because of direct contact harmful liquid, the immunochemical technique objective, efficient, flux is large and related equipment.
Summary of the invention
The object of the present invention is to provide a kind of Full-automatic immunochemical instrument and using method thereof, can avoid damaging in operation, displacement tissue section sample, and, also experimenter can be avoided directly to contact harmful liquid and damage health, simultaneously, improve the reliability of experimental result, repeatability and standardization, realize large flux, high-level efficiency.
For solving the problems of the technologies described above, embodiments of the present invention disclose a kind of Full-automatic immunochemical instrument, comprising: at least three reagent bottles (2), Liquid distribution and liquid discharge structure (3), at least one fluid pipeline (4) and at least one porous reaction ware (5).
Reagent bottle (2) is for holding reactant liquor, washing lotion and liquid waste processing liquid;
Liquid distribution and liquid discharge structure (3) are connected between reagent bottle (2) and fluid pipeline (4), for the liquid in reagent bottle (2) is injected in reaction utensil (5) by fluid pipeline (4), or by the waste liquid in reaction utensil (5) by fluid pipeline (4) sucking-off.
Embodiments of the present invention also disclose a kind of using method of Full-automatic immunochemical instrument, for Full-automatic immunochemical instrument as described above, comprise the following steps:
Reactant liquor is joined in reaction utensil, fully after reaction, by waste liquid sucking-off from reaction utensil;
Washing lotion is joined in reaction utensil, fully after cleaning, by waste liquid sucking-off from reaction utensil;
If immunochemical procedures all completes not yet, then next reactant liquor is joined in reaction utensil.
Compared with prior art, the key distinction and effect thereof are embodiment of the present invention:
Compared with manual immunochemical procedures process pure in prior art, this Full-automatic immunochemical instrument can realize overall process automatic liquid-feeding and sucking-off waste liquid, the mode in ware hole is not changed by changing liquid, frequent moving tissue section sample and damage is caused to it in operation can be avoided, prevent the displacement of sample between different ware hole, avoid experimenter directly contact noxious material and damage health, improve conventional efficient, increase experiment flux, reduce experimental error.
Further, Liquid distribution and liquid discharge structure and fluid pipeline can move up and down, and reduce when liquid feeding and imbibition, raising, making more convenient to operate when changing reaction utensil.
Further, Liquid distribution and liquid discharge structure and fluid pipeline can all around move, and can realize batch processing to the sample matrix such as histotomy or cultured cell, efficiency is higher.
Further, in general, the toxicity of waste liquid is all very strong, very large to the harm of human body and physical environment, after carrying out harmless treatment, can realize qualified discharge, safety and environmental protection by waste collection and harmless treating apparatus to waste liquid.
Further, 4 degrees Celsius of constant temperature and the environment of lucifuge.4 degrees Celsius of constant temperature, can ensure variously to prepare stand-by hydraulic fluid and be in a kind of stable state, extend the expiration date; Under the condition that this kind of low temperature is constant, the speed that immune response can be made to carry out slowly, in conjunction with special, to reduce background local; Can ensure that this Full-automatic immunochemical instrument is under any natural conditions, can work for a long time, reactant liquor and sample can keep optimum performance.The environment of lucifuge is that fluorescent reagent is to the requirement of light in order to meet when carrying out Immunofluorescence Reactions.
Further, the albumen in tissue section sample self is electronegative, after adding positive electricity to reaction utensil, positive and negatively to attract each other, thus ensures that tissue section sample can not be sucked away when sucking-off waste liquid.
Accompanying drawing explanation
Fig. 1 is the structural representation of a kind of Full-automatic immunochemical instrument in first embodiment of the invention;
Fig. 2 is the schematic flow sheet of the using method of a kind of Full-automatic immunochemical instrument in second embodiment of the invention.
Embodiment
In the following description, many ins and outs are proposed in order to make reader understand the application better.But, persons of ordinary skill in the art may appreciate that even without these ins and outs with based on the many variations of following embodiment and amendment, also can realize each claim of the application technical scheme required for protection.
For making the object, technical solutions and advantages of the present invention clearly, below in conjunction with accompanying drawing, embodiments of the present invention are described in further detail.
First embodiment of the invention relates to a kind of Full-automatic immunochemical instrument.Fig. 1 is the structural representation of this Full-automatic immunochemical instrument.
Specifically, as shown in Figure 1: this Full-automatic immunochemical instrument comprises: at least three reagent bottle 2, Liquid distribution and liquid discharge structure 3, at least one fluid pipeline 4 and at least one porous reaction ware 5.
Reagent bottle 2 is for holding reactant liquor, washing lotion and liquid waste processing liquid.
At least three reagent bottles 2, one of them is used for holding washing lotion, other to be used for holding in immunochemical procedures process used various reactant liquors, specifically, various reactant liquor comprises: various antibody as various primary antibodie, various two anti-, three to resist, and positive, negative controls, reaction substrate---diaminobenzidine (DAB), hydrogen peroxide etc.; Washing lotion is used for cleaning samples such as the histotomy after reacted in reactant liquor or cultured cells, and in practical operation, modal washing lotion is exactly phosphate buffer, also can comprise the washing lotion of other kind in addition.
Liquid distribution and liquid discharge structure 3 are connected between reagent bottle 2 and fluid pipeline 4, for being injected in reaction utensil 5 by fluid pipeline 4 by the liquid in reagent bottle 2, or by the waste liquid in reaction utensil 5 by fluid pipeline 4 sucking-off.In operation, Liquid distribution and liquid discharge structure 3 utilize motor or other power, realize the operation of accurate liquid feeding or drawing liquid.Whole operating process is by programmed control, and program can be write.
As shown in Figure 1, each reacting hole of reaction utensil 5 is oral area is circular groove shapes, and mouth diameters is 1 centimetre to 10 centimetres, and reaction utensil can be special, and bottom and the loins in ware hole are mounted with electrode; Also the culture plates such as 6 holes of standard, 12 holes, 24 holes or double dish can be used to replace, thus make the adaptability of this instrument stronger, compatible better.
The arrangement in matrix form between multiple reacting holes of reaction utensil 5, the line space of this matrix is equal with the distance between each fluid pipeline 4.
When reaction utensil 5 has multiple, the also arrangement in matrix form between them.
Liquid distribution and liquid discharge structure 3 and fluid pipeline 4 can move up and down, and reduce when liquid feeding and imbibition, raising, making more convenient to operate when changing reaction utensil.
As shown in Figure 1, Liquid distribution and liquid discharge structure 3 and fluid pipeline 4 are fixed together, and both can move together.By movement, the imbibition mouth of pipe of fluid pipeline 4 end can be deep in each reacting hole of reaction utensil 5 and carry out liquid feeding and imbibition, also can be lifted away from reaction utensil 5.
Liquid distribution and liquid discharge structure 3 and fluid pipeline 4 also can all around move, and like this, can realize batch processing to reaction utensil hole matrix, efficiency is higher.
As shown in Figure 1, according to actual needs, fluid pipeline 4 can be one, also can be many. when fluid pipeline 4 is for many, they can be arranged in a row equally spacedly, when liquid feeding or imbibition, the reacting hole one_to_one corresponding of each fluid pipeline 4 and a certain row in reaction utensil 5 matrix or each reaction utensil 5 in certain a line.
Certainly, in some other embodiment of the present invention, when the quantity of fluid pipeline 4 is abundant, many fluid pipelines 4 also can be arranged in matrix form, the reacting hole one_to_one corresponding of each fluid pipeline 4 in this matrix and each reaction utensil 5 in reaction utensil 5 matrix.
In addition, also comprising: waste collection and harmless treating apparatus 6, for collecting the waste liquid of sucking-off from reaction utensil 5, and harmless treatment being carried out to waste liquid.In general, the toxicity of waste liquid is all very strong, very large to the harm of human body and physical environment, after carrying out harmless treatment, can realize qualified discharge, safety and environmental protection by this waste collection and harmless treating apparatus 6 pairs of waste liquids.
In this waste collection and harmless treating apparatus 6, experimental procedure starts the first step and just automatically adds liquid waste processing liquid in advance.In concrete operating process, can according to used different reactant liquor, waste collection with automatically add corresponding liquid waste processing liquid in harmless treating apparatus 6 in advance and carry out harmless treatment.Such as, when reactant liquor is diaminobenzidine (DAB), in waste collection and harmless treating apparatus 6, adds hydrogen peroxide or 84 thimerosals etc. in advance just can realize harmless treatment to the DAB of severe toxicity containing the material of active oxygen.
Also comprise: casing 1, in this casing 1 be 4 degrees Celsius of constant temperature and the confined space of lucifuge, Full-automatic immunochemical instrument is arranged in this confined space.4 degrees Celsius of constant temperature and the environment of lucifuge, can ensure that this Full-automatic immunochemical instrument is under any natural conditions, can work for a long time, and reactant liquor and the sample such as histotomy, cultured cell can keep optimum performance.
4 degrees Celsius of constant temperature, can ensure variously to prepare stand-by hydraulic fluid and be in a kind of stable state, extend the expiration date; Under the condition that this kind of low temperature is constant, the speed that immune response can be made to carry out slowly, in conjunction with special, reduce local background signal; The environment of lucifuge is that fluorescent reagent is to the requirement of light in order to meet when carrying out Immunofluorescence Reactions.
When Liquid distribution and liquid discharge structure 3 pass through the waste liquid in fluid pipeline 4 sucking-off reaction utensil 5, can allow to stay a part of waste liquid, do not inhale too dry, in order to avoid the sample such as histotomy or cultured cell is sucked away, also can prevent histotomy or cell sample from dehydrating simultaneously, the effect of impact operation, can solve the residual problem of waste liquid by utilizing the washing lotion method of washing several times more.
In addition, in order to avoid in the process of imbibition, the sample such as histotomy or cultured cell is siphoned away simultaneously, when Liquid distribution and liquid discharge structure 3 pass through the waste liquid in fluid pipeline 4 sucking-off reaction utensil 5, should inhale at leisure, the imbibition mouth of pipe of fluid pipeline 4 end also should be tried one's best carefully.
Also comprise: be arranged on reaction utensil 5 top electrode 7, specifically, the bottom in ware hole and loins are mounted with electrode 7, this electrode 7, add positive electricity for during waste liquid in sucking-off reaction utensil 5 to reaction utensil 5.Albumen in tissue section sample self is electronegative, after adding positive electricity to reaction utensil 5, positive and negatively to attract each other, thus ensures that tissue section sample can not be sucked away when sucking-off waste liquid.
Also comprise: shaking table 8, screens established by the top panel of this shaking table 8, and the corresponding reaction utensil 5 of each screens or Tissue Culture Dish, culture plate, for shaking reaction utensil, Tissue Culture Dish or culture plate.
There is screens above this shaking table 8, Tissue Culture Dish, culture plate or special reaction utensil position can be fixed.
By the shake of shaking table 8, liquid can be made fully to mix, avoid the samples such as the multiple histotomy in reaction utensil 5 or cultured cell to be pasted together, with make the sample such as histotomy or cultured cell and reactant liquor fully, uniform contact, ensure to hatch balanced consistent.
Whole immunochemical procedures process is by programmed control, and program can be write.
As shown in Figure 1, this Full-automatic immunochemical instrument also comprises program control instrument 9 and display screen 10, pre-set program, the possibility of newly establishing program also can be provided, full automatic working can be carried out to this Full-automatic immunochemical instrument by program control instrument 9, accurately, convenient, and avoid experimenter and directly contact the risk that noxious material brings.By display screen 10, the information such as the state can clearly observing immunochemical procedures, the reactant liquor used and the time needed for reaction, so that experimenter grasps experiment process.
In addition, be appreciated that program control instrument 9 and display screen 10 can not have yet in some other embodiment of the present invention.
Compared with manual immunochemical procedures process pure in prior art, this Full-automatic immunochemical instrument can realize overall process automatic liquid-feeding and sucking-off waste liquid, the mode in ware hole is not changed by changing liquid, frequent moving tissue section sample and damage, displacement are caused to it in operation can be avoided, and, also experimenter can be avoided directly to contact noxious material and damage health, increase experiment flux, improve conventional efficient, reduce experimental error.
Second embodiment of the invention relates to a kind of using method of Full-automatic immunochemical instrument.Fig. 2 is the schematic flow sheet of the using method of this Full-automatic immunochemical instrument.The using method of this Full-automatic immunochemical instrument is used for Full-automatic immunochemical instrument mentioned above
Specifically, as shown in Figure 2, the using method of this Full-automatic immunochemical instrument comprises the following steps:
In step 201, liquid waste processing liquid is joined in waste collection and harmless treating apparatus, reactant liquor is joined in reaction utensil.
After this step 202 is entered, fully after reaction, by waste liquid sucking-off from reaction utensil.
After this enter step 203, washing lotion is joined in reaction utensil, fully after cleaning, by waste liquid sucking-off from reaction utensil.
In the present embodiment, preferably, step 203 performs three times.
By waste liquid from reaction utensil during sucking-off, retain a part of waste liquid, do not inhale too dry, the object done like this, the sample such as histotomy or cultured cell while sucking-off waste liquid can be avoided to be siphoned away on the one hand simultaneously, also can prevent from the other hand inhaling too dry after, the sample such as histotomy or cultured cell dehydrates, the effect of impact operation.In the present embodiment, the washing lotion method of washing several times can be added by utilization more and solve the residual problem of waste liquid, in the present embodiment, preferably, add washing lotion and wash three times.
Certainly, this be one of the present invention preferred embodiment, in some other embodiment of the present invention, also can only cleaning once, twice, or clean more number of times, as four times, five times etc.
After this enter step 204, judge whether immunochemical procedures all completes.
If so, then process ends; If not, then enter step 205, next reactant liquor is added in reaction utensil.
If immunochemical procedures all completes not yet, then next reactant liquor is joined in reaction utensil.
After this again step 202 is got back to.
Present embodiment is the method embodiment corresponding with the first embodiment, and present embodiment can be worked in coordination with the first embodiment and be implemented.The relevant technical details mentioned in first embodiment is still effective in the present embodiment, in order to reduce repetition, repeats no more here.Correspondingly, the relevant technical details mentioned in present embodiment also can be applicable in the first embodiment.
Each method embodiment of the present invention all can realize in modes such as software, hardware, firmwares.No matter the present invention realizes with software, hardware or firmware mode, instruction code can be stored in the addressable storer of computing machine of any type (such as permanent or revisable, volatibility or non-volatile, solid-state or non-solid, fixing or removable medium etc.).Equally, storer can be such as programmable logic array (Programmable Array Logic, be called for short " PAL "), random access memory (Random Access Memory, be called for short " RAM "), programmable read only memory (Programmable Read Only Memory, be called for short " PROM "), ROM (read-only memory) (Read-Only Memory, be called for short " ROM "), Electrically Erasable Read Only Memory (Electrically Erasable Programmable ROM, be called for short " EEPROM "), disk, CD, digital versatile disc (Digital Versatile Disc, be called for short " DVD ") etc.
Although by referring to some of the preferred embodiment of the invention, to invention has been diagram and describing, but those of ordinary skill in the art should be understood that and can do various change to it in the form and details, and without departing from the spirit and scope of the present invention.
Claims (9)
1. a Full-automatic immunochemical instrument, it is characterized in that, comprising: at least three reagent bottles (2), Liquid distribution and liquid discharge structure (3), at least one fluid pipeline (4) and at least one porous reaction ware (5);
Reagent bottle (2) is for holding reactant liquor, washing lotion and liquid waste processing liquid;
Liquid distribution and liquid discharge structure (3) are connected between reagent bottle (2) and fluid pipeline (4), for the liquid in reagent bottle (2) is injected in the reacting hole of reaction utensil (5) by fluid pipeline (4), or by the waste liquid in reaction utensil (5) by fluid pipeline (4) sucking-off;
This Full-automatic immunochemical instrument also comprises: be arranged on the electrode (7) on reaction utensil (5), and this electrode (7) adds positive electricity for during waste liquid in sucking-off reaction utensil (5) to reaction utensil (5).
2. Full-automatic immunochemical instrument according to claim 1, is characterized in that, each reacting hole of described reaction utensil (5) is oral area is circular groove shapes, and mouth diameters is 1 centimetre to 10 centimetres;
The arrangement in matrix form between multiple reacting holes of reaction utensil (5), the line space of this matrix is equal with the distance between each fluid pipeline (4).
3. Full-automatic immunochemical instrument according to claim 2, is characterized in that, described Liquid distribution and liquid discharge structure (3) and fluid pipeline (4) can move up and down.
4. Full-automatic immunochemical instrument according to claim 3, is characterized in that, described Liquid distribution and liquid discharge structure (3) and fluid pipeline (4) can all around move.
5. Full-automatic immunochemical instrument according to claim 4, it is characterized in that, also comprise: waste collection and harmless treating apparatus (6), automatically treating fluid is added in advance in this waste collection and harmless treating apparatus (6), for collecting the waste liquid of sucking-off from reaction utensil (5), and harmless treatment is carried out to waste liquid.
6. Full-automatic immunochemical instrument according to claim 5, it is characterized in that, also comprise: casing (1), in this casing (1) be 4 degrees Celsius of constant temperature and the confined space of lucifuge, described Full-automatic immunochemical instrument is arranged in this confined space.
7. Full-automatic immunochemical instrument according to claim 6, it is characterized in that, also comprise: shaking table (8), screens established by the top panel of this shaking table (8), the corresponding reaction utensil (5) of each screens, for shaking reaction utensil (5).
8. a using method for Full-automatic immunochemical instrument, for the Full-automatic immunochemical instrument such as according to any one of claim 1 to 7, is characterized in that, comprises the following steps:
Reactant liquor is joined in reaction utensil, fully after reaction, by waste liquid sucking-off from reaction utensil;
Washing lotion is joined in reaction utensil, fully after cleaning, by waste liquid sucking-off from reaction utensil;
If immunochemical procedures all completes not yet, then next reactant liquor is joined in reaction utensil.
9. the using method of Full-automatic immunochemical instrument according to claim 8, is characterized in that, describedly joins in reaction utensil by washing lotion, fully after cleaning, waste liquid step of sucking-off from reaction utensil is performed three times.
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| JP2002189033A (en) * | 2000-12-22 | 2002-07-05 | Furuno Electric Co Ltd | Method and system for dispensing, and tip stocker device |
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| CN101865927A (en) * | 2009-04-17 | 2010-10-20 | 武汉医尔特科技有限公司 | Novel full-automatic blood type analyzer |
| JP2011153940A (en) * | 2010-01-28 | 2011-08-11 | Hitachi High-Technologies Corp | Automatic analyzer |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN2170520Y (en) * | 1993-06-16 | 1994-06-29 | 陆雪龙 | Automatic operating apparatus for determining radioimmunity |
| CN2444230Y (en) * | 2000-05-16 | 2001-08-22 | 陈智 | Immunohistochemistry apparatus |
| JP2002189033A (en) * | 2000-12-22 | 2002-07-05 | Furuno Electric Co Ltd | Method and system for dispensing, and tip stocker device |
| CN2859530Y (en) * | 2005-12-29 | 2007-01-17 | 王维锋 | Protein mark immunological automatic detection |
| CN101865927A (en) * | 2009-04-17 | 2010-10-20 | 武汉医尔特科技有限公司 | Novel full-automatic blood type analyzer |
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