CN102321535A - Method and system for automatic control of processing and sequencing of biological samples - Google Patents
Method and system for automatic control of processing and sequencing of biological samples Download PDFInfo
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- CN102321535A CN102321535A CN201110258910A CN201110258910A CN102321535A CN 102321535 A CN102321535 A CN 102321535A CN 201110258910 A CN201110258910 A CN 201110258910A CN 201110258910 A CN201110258910 A CN 201110258910A CN 102321535 A CN102321535 A CN 102321535A
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Abstract
The invention relates to the field of genetic engineering, and provides a method and a system for automatic control of processing and sequencing of biological samples. The method comprises the following steps that: A, the preparation of biological samples is controlled, and a response system is generated and is arranged in a gene sequencing instrument; B, the gene sequencing instrument is controlled to allow a reagent to be introduced into the response system, and the temperature of the response system is regulated; C, the response system is controlled to move in the gene sequencing instrument, and an image aquiring position is determined; and D, markers carried by nucleotide in the response system are excitated to emit light, and image signals are obtained at the image aquiring position. The system comprises a sample control unit, a response control unit, a positioning control unit and an image aquiring control unit, which are respectively used for executing the steps in the method. In the method, the preparation and the loading of the biological samples are automatically controlled, and the sequencing process of the gene sequencing is automatically controlled, accordingly, the stability and the efficiency of the sequencing process are improved, and the accuracy of sequencing results is improved.
Description
Technical field
The present invention relates to the genetically engineered field, more particularly, relate to a kind of biological sample is handled and robotization control is carried out in order-checking method and system.
Background technology
Initial gene sequencing technology is carried out through manual operations, comprises the dideoxy chain termination of Sanger invention, and the chemical degradation method of Maxam and Gilbert invention.Because efficient hand-manipulated is lower, and the human operational error is taken place easily, therefore utilize gene sequencer to check order has become the main flow of sequencing technologies at present.
In present stage; If will check order or data analysis to gene fragment, need at first from biont, for example histocyte, bacterium etc. are extracted genetic material (RNA); And utilize to transcribe and synthesize and obtain dna fragmentation, the general dna fragmentation that obtains by this method all is a spot of.Then dna fragmentation is increased; For example can utilize polymerase chain reaction (Polymerase Chain Reaction; PCR), transcribe method etc. and increase, obtain a large amount of gene fragments, and then check order or data analysis based on the gene fragment after the amplification.Therefore before formal order-checking, need do the specimen preparation work in a large amount of early stages, hand-manipulated carrying out all adopted in these work all the time, and the sample for preparing is set in the sequenator also is through manually, can't realize robotization.Just there is the problem of two aspects in this: carrying out specimen preparation need spend the plenty of time by hand on the one hand, and manual operations is difficult to guarantee the quality of specimen preparation each time on the other hand, and will directly influence order-checking can carry out smoothly, also can influence the accuracy of sequencing result.
In addition; Arrived the order-checking stage; With regard to the control of gene sequencer itself; The order-checking process of gene sequencer is made up of operations such as a series of machineries, electronic communication, biology, chemistry, optics at present, and these operations are performed by assembly corresponding in the gene sequencer respectively, substituted simple manual operations.But also face following problem: on the one hand, because gene sequencing is very high to the requirement of precision, belong to nano level, the operation of any one assembly deviation occurs and all can cause sequencing result undesirable; On the other hand, the concrete steps that the whole measuring program process relates to are very loaded down with trivial details, need to carry out Collaboration between each assembly in the gene sequencer.That is to say that the order-checking process not only requires in the gene sequencer each assembly to carry out operations accurately and rapidly, also requires to carry out between each assembly good cooperation.In concrete the application; The factor that relates to when gene sequencer checks order is very complicated; Comprise many-sided controls such as reagent dosage and type, temperature of reaction, time, cleanliness factor, nano-grade displacement, focus adjustment, luminous intensity, light path adjusting, time shutter calculating, image takings; Therefore and the requirement of each aspect is very high, will guarantee that the order-checking process carries out smoothly, and difficulty is very big.
Only the control with reagent dosage and type describes; Since in the gene sequencing process to the control of reagent dosage generally at micro updating; And need carry out repeatedly the absorption importing of various dose in different step of reaction; Add that each selected types of agents all possibly there are differences, and has therefore proposed higher requirement to the dosage of reagent, the assurance of type.Draw if carry out reagent, or the MC instrument carries out reagent and draw, all have following problem: be difficult to accurately control dosage on the one hand, and the nuance of dosage can cause different biochemical reaction results, also will directly influence sequencing result by manual operation; On the other hand, artificial participation need accurately judge all ingredients type of reaction different steps, even if the error on annelet will cause the biochemical reaction failure, makes the whole measuring program process fail totally.In addition; Because up to drawing sequencing result, each stage all needs certain cycle to the order-checking process from specimen preparation, last appearance, order-checking, data analysis, if there is error in the control of mentioned reagent dosage and type; Manual operation can't be monitored; Can not be in subsequent process error correction in time, also be difficult to find the basic reason of order-checking procedure failure even if learn final sequencing result failure, also wasted great amount of time and expensive reagent.
Factor except reagent dosage and type; Other various factorss; Comprise aforesaid temperature of reaction, time, cleanliness factor, nano-grade displacement, focus adjustment, luminous intensity, light path adjusting, time shutter calculating, image taking etc., all have above-mentioned similar situation, if there is not the system of robotization to operate; The whole measuring program process will be difficult to smooth expansion, and it is just more difficult to want to stablize, obtain apace accurately sequencing result.
Therefore need a kind of the method that biological sample is handled and the order-checking process of gene sequencer carried out robotization control to be solved an above-mentioned difficult problem, guarantee the order-checking process can be smoothly, carry out expeditiously, and improve the accuracy of sequencing result.
Summary of the invention
The object of the present invention is to provide a kind ofly, be intended to improve the stability and the efficient of order-checking process, improve the accuracy of sequencing result simultaneously the method and system that biological sample is handled and robotization control is carried out in order-checking.
In order to realize goal of the invention, said the system that biological sample is handled and robotization control is carried out in order-checking is comprised sample control unit, reaction control unit, Positioning Control Unit, adopts the figure control unit; Said sample control unit is used to control the preparation of biological sample, and the formation reaction system also is set to it in gene sequencer; Said reaction control unit is used for the controlling gene sequenator reagent is imported reaction system, and in the order-checking process temperature of conditioned reaction system; Said Positioning Control Unit is used for controlling reaction system the moving of gene sequencer, and confirms to adopt the figure position; It is luminous that the said figure of adopting control unit is used for the affinity tag that provocative reaction system Nucleotide carries, and obtain picture signal in the said figure of adopting position.
Wherein, said sample control unit comprises that system prepares module, goes up the appearance control module; Said system prepares module and prepares gene fragment storehouse to be measured according to biological sample, and is processed into the required reaction system of order-checking; The said appearance control module that goes up is set in the gene sequencer allocation really with reaction system.
Wherein, said reaction control unit comprises reagent control module, temperature control module; Said reagent control module is used for the controlling gene sequenator to be selected reagent, and draws corresponding reagent, imports reaction system; Said temperature control module is used for the temperature of reaction system is controlled at the required temperature of reaction.
Wherein, said Positioning Control Unit comprises displacement module, focus module; Said displacement module is used for the current position of detection reaction system at gene sequencer, and controls it and move to its target location of place on plane; Said focus module is used for the focus adjustment of controlling gene sequenator, and that confirms reaction system adopts the figure position.
Wherein, said focus module is through regulating the distance between microscope and the reaction small chamber, and the position that sharpness is best is confirmed as and adopted the figure position.
Wherein, the said figure of adopting control unit comprises excitation module, photo module, image access module; Said excitation module is used to control the excitation light irradiation reaction system of specific wavelength, makes the affinity tag that Nucleotide carries in the reaction system luminous; Said photo module is confirmed the time shutter, and adopts the said time shutter that reaction system is taken pictures, and obtains picture signal; Said image access module and photo module communicate, and are used to preserve the picture signal of obtaining.
In order to realize goal of the invention better; Said to biological sample is handled and robotization control is carried out in order-checking method; It is characterized in that, said method comprising the steps of: the preparation of A. control biological sample, the formation reaction system also is set to it in gene sequencer; B. the controlling gene sequenator imports reaction system with reagent, and the temperature of conditioned reaction system; C. control reaction system moving in gene sequencer, and confirm to adopt the figure position; D. the affinity tag that Nucleotide carries in the provocative reaction system is luminous, and obtains picture signal in the said figure of adopting position.
Wherein, said steps A comprises: A1. prepares gene fragment storehouse to be measured according to biological sample, and is processed into the required reaction system of order-checking; A2. said reaction system is set in the gene sequencer allocation really.
Wherein, said step B comprises: B1. controlling gene sequenator is selected reagent, and draws corresponding reagent, imports reaction system; B2. the temperature with reaction system is controlled at the required temperature of reaction.
Wherein, said step C comprises: the current position of C1. detection reaction system in gene sequencer, and control it and move to its target location of place on plane; C2. the focus adjustment of controlling gene sequenator, that confirms reaction system adopts the figure position.
Wherein, said step C2 comprises: through regulating the distance between microscope and the reaction small chamber, the position that sharpness is best is confirmed as and is adopted the figure position.
Wherein, said step D comprises: the excitation light irradiation reaction system of D1. control specific wavelength makes the affinity tag that Nucleotide carries in the reaction system luminous; D2. confirm the time shutter, and adopt the said time shutter that reaction system is taken pictures, obtain picture signal; D3. preserve the picture signal of obtaining.
By on can know that the present invention is through to the preparation of biological sample and go up appearance and carry out robotization control, and the order-checking process of gene sequencing is carried out robotization control, not only improved the stability and the efficient of order-checking process, and improved the accuracy of sequencing result.
Description of drawings
Fig. 1 is that the present invention handles biological sample and the system architecture synoptic diagram that robotization is controlled is carried out in order-checking;
Fig. 2 is system 1 structural representation in one embodiment among Fig. 1;
Fig. 3 is sample control unit 400 structural representation in one embodiment among Fig. 2;
Fig. 4 is reaction control unit 100 structural representation in one embodiment among Fig. 2;
Fig. 5 is Positioning Control Unit 200 structural representation in one embodiment among Fig. 2;
Fig. 6 adopts figure control unit 300 structural representation in one embodiment among Fig. 2;
Fig. 7 carries out the method flow diagram that robotization is controlled to biological sample processing and order-checking in the one embodiment of the invention;
Fig. 8 is a step S0 method flow diagram in one embodiment among Fig. 7;
Fig. 9 is a step S1 method flow diagram in one embodiment among Fig. 7;
Figure 10 is a step S2 method flow diagram in one embodiment among Fig. 7;
Figure 11 is a step S3 method flow diagram in one embodiment among Fig. 7.
Embodiment
In order to make the object of the invention, technical scheme and advantage clearer,, the present invention is further elaborated below in conjunction with accompanying drawing and embodiment.
Fig. 1 shows the present invention the system architecture that robotization is controlled is carried out in biological sample processing and order-checking; This system comprises system 1; With coupled at least one stylobate because of sequenator, gene sequencer 2 as shown in the figure, gene sequencer 3...... gene sequencer N.Should be noted that the annexation between each equipment or the module is the needs for clear its information interaction of explaination and control process in all diagrams of the present invention, therefore should be regarded as control relation in logic, and should not be limited to physical connection or wireless connections.Need to prove that in addition the signalling methods between each functional module can be taked multiple, protection scope of the present invention should not be defined as the signalling methods of certain particular type.Wherein:
(1) system 1 is used for communicating because of sequenator with at least one stylobate, each corresponding assembly in its each functional module difference controlling gene sequenator, perhaps, the order-checking process of controlling gene sequenator.Mainly comprise: the controlling gene sequenator imports reaction system with reagent, and the temperature of conditioned reaction system; Control reaction system moving in gene sequencer, and confirm to adopt the figure position; The affinity tag that Nucleotide carries in the provocative reaction system is luminous, and obtains picture signal in the above-mentioned figure of adopting position.Should be noted that above-mentioned mode is applicable to various types of gene sequencers, so the protection domain of control method and system should not receive the restriction of this body structure of gene sequencer among the present invention.About the particular content of system 1, will in embodiment thereafter, set forth in detail.
(2) gene sequencer N is made up of a plurality of assemblies, respectively with system 1 in each functional module corresponding, accept and carry out the instructions of these functional modules, thereby collaboratively accomplish order-checking.These assemblies comprise: be used to the assembly drawing reagent and import reaction system; Be used for assembly that the temperature of reaction system is regulated; The assembly that accommodates reaction system and can in gene sequencer, move; Be used to confirm to adopt the assembly of figure position, be used to import the assembly of exciting light, be used to assembly of gathering picture signal etc.Should be noted that dissimilar gene sequencers has different intrawares, perhaps the external expressive form of intraware is different, but the function that is realized is consistent, and protection scope of the present invention should not receive the restriction of these factors.Also should be noted that not necessarily complete independence between each assembly, realize that each assembly of difference in functionality may relate to one or more identical parts.Constitute about the part assembly among the gene sequencer N; But application reference people's publication: application number is CN200810132008.8; Denomination of invention is " sequencing reaction small chamber, gene sequencing reaction bench and a gene sequencing system ", and the present invention also will specifically set forth in embodiment thereafter.
Need to prove that about reaction system this reaction system comprises a plurality of separate reactions drips, each reaction is dripped and is comprised a plurality of copy number purposes dna fragmentation to be measured, generally is up to ten million even more than one hundred million orders of magnitude in the present invention.In the order-checking process; Dna fragmentation to be measured in the reaction system will combine with the Nucleotide that has affinity tag; This affinity tag can receive the light source activation of specific wavelength, makes the microballon that has this affinity tag luminous, so just can gather picture signal; And handle and analyze according to picture signal, obtain gene order information.
Fig. 2 shows the structure of system 1 in first embodiment among Fig. 1, comprises sample control unit 400, reaction control unit 100, Positioning Control Unit 200, adopts figure control unit 300.Wherein:
(1) sample control unit 400 is used to control the preparation of biological sample, and the formation reaction system also is set to it in gene sequencer.Need to prove, should be regarded as control relation in logic in sample control unit 400 and the system 1 between other unit, and should not be limited to physical connection or wireless connections.The hardware components that sample control unit 400 is controlled may reside in gene sequencer inside, also can should not be regarded as the restriction to protection domain of the present invention in the outside independent existence of gene sequencer.
For example; Gene sequencer inside has a plurality of assemblies corresponding with sample control unit 400; Sample control unit 400 is actually the operation through the control section assembly; The biological sample that is collected is prepared into required reaction system, and, position suitable in the gene sequencer is installed or be set to this reaction system through controlling the operation of another part assembly.Examination is that example is explained above-mentioned control process with one type gene sequencer; In this concrete situation; Comprise the following assembly corresponding with sample control unit 400 in the gene sequencer: (1) library prepares assembly, is used for biological sample is prepared into gene fragment storehouse to be measured; (2) reaction system prepares assembly, is used for gene fragment to be measured is prepared into the required reaction system of order-checking; (3) mechanical manipulator is installed to position suitable in the gene sequencer with reaction system, generally is mounted in the sample table.In this case; The control process of sample control unit 400 is: prepare assembly through the control library biological sample is prepared into gene fragment storehouse to be measured; Further control reaction system again and prepare assembly gene fragment is prepared into the required reaction system of order-checking, control mechanical manipulator then and install.About the specific functional modules and the mode of sample control unit 400, will in embodiment thereafter, set forth in detail.
(2) reaction control unit 100 is used for the controlling gene sequenator reagent is imported reaction system, and in the order-checking process temperature of conditioned reaction system.
In the present invention; Gene sequencer inside has and the corresponding a plurality of assemblies of reaction control unit 100; Reaction control unit 100 is actually the operation through the control section assembly; Reagent is imported reaction system, and, realize adjusting temperature of reaction system through controlling the operation of another part assembly.Examination is that example is explained above-mentioned control process with one type gene sequencer, and in this concrete situation, comprise in the gene sequencer and the corresponding following assembly of reaction control unit 100: (1) reagent platform is used to place or holds the multiple reagent of supply drawing; (2) mechanical manipulator is used for selecting suitable reagent in different step of reaction; (3) pump and flexible pipe are used to draw selected reagent, and wherein flexible pipe is fixed on the mechanical manipulator; (4) sample table, it comprises reaction small chamber, is used to hold reaction system; (5) temperature controller links to each other with reaction small chamber, and it comprises the TP that is used for the little chambers temp of detection reaction, and to the heat riser of reaction small chamber heating.In this case; The control process of reaction control unit 100 is: through control mechanical manipulator, pump and flexible pipe, reagent is imported reaction system, and through the TP detected temperatures; And control heat riser and heat, thereby the temperature of conditioned reaction system to reaction small chamber.Should be noted that; For dissimilar gene sequencers; The type of the assembly that it is included, structure or quantity possibly there are differences, but the control process of reaction control unit 100 is consistent on ultimate principle, so protection domain should not receive the restriction of above-mentioned factor.About the specific functional modules and the mode of reaction control unit 100, will in embodiment thereafter, set forth in detail.
(2) Positioning Control Unit 200 is used for controlling reaction system the moving of gene sequencer, and confirms to adopt the figure position.
In the present invention; Gene sequencer inside has a plurality of assemblies corresponding with Positioning Control Unit 200; Positioning Control Unit 200 is actually the operation through the control section assembly; Thereby control reaction system moving in gene sequencer, and pass through to control the operation of another part assembly, thus confirm to adopt the figure position.Examination is that example is explained above-mentioned control process with one type gene sequencer, in this concrete situation, comprises as follows the assembly corresponding with Positioning Control Unit 200 in the gene sequencer: (1) sample table; It is the sample table that aforementioned content is mentioned; It comprises reaction small chamber, is used to hold reaction system, as previously mentioned; Not necessarily independent fully between each assembly, realize that each assembly of difference in functionality may relate to one or more identical parts; (2) microscope is used for focusing is regulated.In this case; The control process of Positioning Control Unit 200 is: through moving of control sample table; Make reaction system move to position suitable in the gene sequencer, and through the distance between control microscope and the reaction small chamber, thereby confirm the suitable figure position of adopting.About the specific functional modules and the mode of Positioning Control Unit 200, will in embodiment thereafter, set forth in detail.
(3) adopting figure control unit 300, to be used for the affinity tag that provocative reaction system Nucleotide carries luminous, and obtain picture signal in the above-mentioned figure of adopting position.
In the present invention; Gene sequencer inside has a plurality of assemblies corresponding with adopting figure control unit 300; Adopt figure control unit 300 and be actually operation through the control section assembly; Thereby the affinity tag that Nucleotide carries in the provocative reaction system is luminous, and the operation through control another part assembly, thereby obtains picture signal adopting the figure position.Examination is that example is explained above-mentioned control process with one type gene sequencer, in this concrete situation, comprises as follows the assembly corresponding with adopting figure control unit 300 in the gene sequencer: (1) excitation light source; (2) CCD is used for photographic images.In this case, the control process of adopting figure control unit 300 is: luminous through the control excitation light source, the affinity tag that Nucleotide carries in the provocative reaction system is luminous, and through control CCD photographic images, thereby the picture signal of obtaining.About the specific functional modules and the mode of adopting figure control unit 300, will in embodiment thereafter, set forth in detail.
In concrete the application; Said system shown in Figure 2 utilizes sample control unit 400, reaction control unit 100, Positioning Control Unit 200, adopt in 300 pairs of gene sequencers of figure control unit each corresponding assembly carries out automated operation respectively, and can effectively coordinate different assemblies.What is more important; Every operation has all taken into full account the technical characterstic in each stage of gene sequencing; Comprise the control of reaction 100 pairs of reagent dosage of control unit and type, temperature of reaction, time, cleanliness factor etc.; The control of 200 pairs of nano-grade displacements of Positioning Control Unit, focus adjustment etc., the control of adopting 300 pairs of luminous intensities of figure control unit, light path adjusting, time shutter calculating, image taking etc. is all operated according to strict index accurately.Therefore system of the present invention can increase substantially the stability and the efficient of order-checking process, and the accuracy of sequencing result.
Fig. 3 shows sample control unit 400 structure in one embodiment among Fig. 2, comprises that system prepares module 401, goes up appearance control module 402.Wherein:
(1) system prepares module 401 and prepares gene fragment storehouse to be measured according to biological sample, and generates the required reaction system of order-checking.In the present invention, the specimen preparation process of reaction system realizes through a water in oil unique DNA fragment amplification system.In water in oil system, comprise a large amount of separate reactions and drip, each reaction is dripped and is comprised a microballon, is combined with dna fragmentation to be measured on it.Through water in oil system is carried out pcr amplification, each magnetic bead will combine a plurality of copy number target DNA fragments after the amplification, and these fragments all come from same dna profiling to be measured.Certainly, the present invention also can also can carry out subsequent operations to unit molecule without pcr amplification, just behind pcr amplification, can strengthen the order-checking signal, improves sequencing quality.
Situation with the gene sequencer assembly described among Fig. 2 is an example; This preparation process is to utilize system to prepare that library in 401 pairs of gene sequencers of module prepares assembly, reaction system prepares assembly and controls; Specifically comprise: (a) system prepares module 401 transmission instructions and prepares assembly to the library; Control this assembly and under different experiment conditions, biological sample is carried out a series of processing, make biological sample accomplish the transformation of several stages, obtain gene fragment storehouse to be measured.What need explanation is that (b) system prepares module 401 transmission instructions and prepares assembly to reaction system, controls the mixing that this assembly carries out oil phase, water, generates water in oil reaction system under certain condition.
In above-mentioned (a), as if the type of the biological sample that obtains at first, stage of living in difference, system prepares module 401 will corresponding multiple different processing mode.In one embodiment, if the biological sample that obtains at first is histocyte, bacterium etc., system prepares module 401 and will at first control the library and prepare assembly and from these histocytes, bacterium, extract RNA so, and transcribes the synthetic initial dna fragmentation that obtains.Then initial dna fragmentation is attached on the microballon; To be combined in a plurality of initial dna fragmentation enzyme on the microballon again and be cut into a plurality of DNA labels of equal in length; Then each DNA label is connected one section universal sequence; At last each the DNA label that is connected with universal sequence is increased, obtain gene fragment storehouse to be measured.In another embodiment; If the biological sample that obtains at first has been through extracting, transcribe the synthetic initial dna fragmentation that obtains; System prepares 401 of modules control library and prepares assembly initial dna fragmentation is attached on the microballon so, will be combined in a plurality of DNA labels that a plurality of initial dna fragmentation enzyme on the microballon is cut into equal in length again, then each DNA label is connected one section universal sequence; At last each the DNA label that is connected with universal sequence is increased, obtain gene fragment storehouse to be measured.
In above-mentioned (b), the preparation of water-in-oil reaction system has multiple mode.In one embodiment, system prepares module 401 at first to be controlled reaction system and prepares assembly and set up the microballon comprise after the modification at interior PCRmix aqueous phase system, for example sets up following aqueous phase system: (1) 10 * PCR buffer, 15 μ L; (2) dNTP, 3 μ L; (3) dna profiling, 2 μ L; (4) microballon after primer combines, 5 μ L; (5) Taq enzyme, 9 μ L; (6) Mg2+, 3 μ L; (7) ddH
2O, 114 μ L; (8) preferred adding can add a pair of little primer of fast response toggle speed, each 0.75 μ L of two ends, upstream and downstream in aqueous phase system.System prepares module 401 to be controlled reaction system then and prepares assembly and set up the oil phase system that oils solution constitutes, and then aqueous phase system is injected into the oil phase system, and shaking mixing under certain condition becomes emulsion.This condition is for example shaken frequency 15HZ, time 15s.Above-mentioned water-in-oil reaction system can prepare in multiple container, for example in the test tube.
(2) going up appearance control module 402 is set to reaction system in the gene sequencer.
Situation with the gene sequencer assembly described among Fig. 2 is an example, and this setting up procedure is to utilize the mechanical manipulator of going up in 402 pairs of gene sequencers of appearance control module to control, and reaction system is set in the movable assembly in the gene sequencer.In one embodiment, last appearance control module 402 is sent instruction and is given mechanical manipulator, and the water-in-oil reaction system in the aforesaid test tube is imported a reaction small chamber, reaction small chamber is installed to the specific position of sample table again.
So far, accomplished the specimen preparation of robotization and gone up the appearance operation, then can begin to check order.
Fig. 4 shows reaction control unit 100 structure in one embodiment among Fig. 2, comprises reagent control module 101, temperature control module 102.Wherein:
(1) reagent control module 101 is used for the controlling gene sequenator reagent is selected, and draws corresponding reagent, imports reaction system.
Situation with the gene sequencer assembly described among Fig. 2 is an example; The concrete control process of reagent control module 101 realizes through the serial communication mode; Detailed process is: reagent control module 101 is confirmed the required types of agents of taking of different steps; Send instructions to mechanical manipulator, the control mechanical manipulator moves to reagent position corresponding on the reagent platform, and the flexible pipe that will be fixed on the mechanical manipulator inserts in the reagent; Reagent control module 101 sends instructions to pump, thereby reagent is drawn in the control pump running; Reagent control module 101 sends instructions to pump after being drawn to required reagent, continues the control pump running reagent is squeezed into reaction small chamber.
(2) temperature control module 102 is used for the temperature of reaction system is controlled at the required temperature of reaction.
Situation with the gene sequencer assembly described among Fig. 2 is an example; The concrete control process of temperature control module 102 realizes through the serial communication mode; Detailed process is: temperature control module 102 sends instruction and gives TP; The controlled temperature transmitter detects the temperature of reaction small chamber, and reads temperature detection result t; Be provided with the temperature value T in differential responses stage in the temperature control module 102, after it obtains temperature detection result t, then it compared with the temperature value T that is provided with; Temperature control module 102 is further handled according to comparing result; If t＜T, temperature control module 102 sends instruction and gives the heat riser in the temperature controller, and the heat riser in the control temperature controller starts; Heat to reaction small chamber; If t >=T then need not start the heat riser heating, heat riser is cooled to temperature T automatically through outside atmosphere.
For the gene sequencer assembly under other situations, control principle is consistent, and detailed process possibly there are differences.For example; In the gene sequencer assembly under another kind of situation; The situation of describing in Fig. 2, temperature controller except comprise the TP that is used for the little chambers temp of detection reaction, to the heat riser of reaction small chamber heating, also comprise to reaction small chamber refrigerating heat sink.So in this case, the concrete control process of temperature control module 102 is: temperature control module 102 sends instruction and gives TP, and the controlled temperature transmitter detects the temperature of reaction small chamber, and reads temperature detection result t; Be provided with the temperature value T in differential responses stage in the temperature control module 102, after it obtains temperature detection result t, then it compared with the temperature value T that is provided with; Temperature control module 102 is further handled according to comparing result, if t＜T, temperature control module 102 sends instruction and gives the heat riser in the temperature controller; Heat riser in the control temperature controller starts; To the reaction small chamber heating, if t >=T, temperature control module 102 sends instruction and gives the heat sink in the temperature controller; Heat sink in the control temperature controller starts, and reaction small chamber is freezed.
By on can know; Reagent control module 101 can be controlled types of agents, reagent dosage, reagent transmission speed etc. accurately; Temperature control module 102 can carry out strictness control to temperature detection, temperature setting, heating, refrigeration etc.; Thereby the biochemical reaction process that has guaranteed reaction system carries out smoothly, has also therefore improved stability, efficient and the accuracy of whole measuring program process.
Fig. 5 shows Positioning Control Unit 200 structure in one embodiment among Fig. 2, comprises displacement module 201, focus module 202.Wherein:
(1) displacement module 201 is used for the current position of detection reaction system at gene sequencer, and controls it and move to its target location of place on plane.Among the present invention, displacement module 201 can be controlled moving of reaction system in several ways.
In one embodiment; Be the situation of describing in the earlier figures 2 with Positioning Control Unit 200 corresponding assemblies in the gene sequencer; The concrete control process of displacement module 201 realizes through the serial communication mode: displacement module 201 at first sends instruction and gives sample table; Reading the starting position coordinate of sample table on its plane, place, for example is (X
0, Y
0); Confirm reaction system at its place the target coordinate on plane (X, Y) after, sample table is given in displacement module 201 instruction of redispatching, the control sample table is from (X
0, Y
0) move to target coordinate (X, Y).
(2) focus module 202 is used for the focus adjustment of controlling gene sequenator, and that confirms reaction system adopts the figure position.Among the present invention, what focus module 202 can be confirmed reaction system in several ways adopts the figure position, and the situation with the gene sequencer assembly described among Fig. 2 is that example describes below.
In one embodiment; The concrete control process of focus module 202 realizes through the serial communication mode: focus module 202 is at first sent instruction and is given microscope; The control microscope with the sample table vertical direction on move; Through regulating the distance between microscope and the reaction small chamber, the position that sharpness is best is confirmed as and is adopted the figure position.
In another embodiment, focus module 202 is sent instruction and is given sample table, and the control sample table moves on its place planar vertical direction, and through the distance between conditioned reaction cell and the microscope, the position that sharpness is best is confirmed as and adopted the figure position.
In above-mentioned two embodiment, focus module 202 can be confirmed the position that image definition is best in several ways.For example, can observe the position that mode of comparing confirms that image definition is best, perhaps utilize the arithmetic calculation image definition, utilize algorithm to regulate image definition automatically through the microscope camera lens, or the like.
By on can know; Displacement module 201 can carry out accurate control automatically to the displacement of micro/nano level; Focus module 202 can be carried out accurate control automatically to focus adjustment, definition judgment etc.; Thereby can confirm the best figure position of adopting quickly and accurately, also therefore improve stability, efficient and the accuracy of order-checking process.
Fig. 6 shows and adopts figure control unit 300 structure in one embodiment among Fig. 2, comprises excitation module 301, photo module 302, image access module 303.Wherein:
(1) excitation module 301 is used to control the excitation light irradiation reaction system of specific wavelength, makes the affinity tag that Nucleotide carries in the reaction system luminous.In the present invention, excitation module 301 can make affinity tag luminous in several ways.
In one embodiment, be the situation of describing in the earlier figures 2 with adopting figure control unit 300 corresponding assemblies in the gene sequencer, excitation module 301 is sent instruction and is given excitation light source so, and it is luminous to start excitation light source, makes the reaction small chamber of irradiate light on the sample table.In the present embodiment, the affinity tag that Nucleotide carries on the microballon in the reaction system is a fluorescent marker, and it just can send fluorescence after receiving the light source activation of specific wavelength.
(2) photo module 302 is confirmed the time shutter, and adopts the said time shutter that reaction system is taken pictures, and obtains picture signal.In the present invention, photo module 302 can be obtained picture signal in several ways.
In one embodiment; Be the situation of describing in the earlier figures 2 with adopting figure control unit 300 corresponding assemblies in the gene sequencer; Photo module 302 is at first confirmed suitable exposure time values so, sends instruction then and gives CCD, and control CCD takes fluorogram according to this exposure time values.
(3) image access module 303 communicates with photo module 302, is used to preserve the picture signal of obtaining.Image access module 303 can adopt multiple format picture signal among the present invention.In the aforementioned embodiment; After photo module 302 control CCD take fluorogram; Send to image access module 303; Image access module 303 can adopt special high fidelity visual storage format to preserve fluorogram, also can adopt common image storage format, for example TIFF, EPS, PNG, PSD form etc.Protection scope of the present invention should not receive the restriction of image storage format.
By on can know; Excitation module 301 can accurately be controlled luminous light path of excitation light source etc.; Photo module 302 can accurately be controlled the confirming of exposure time values, image taking etc., and image access module 303 can adopt best picture format memory image signal, thus the quality of the picture signal that has guaranteed to be obtained; Greatly improved the accuracy of sequencing result, and this robotization mode the stability and the efficient of order-checking process have also been improved.
The method flow that the present invention handles biological sample and robotization control is carried out in order-checking is based on system shown in Figure 1, and this system comprises that system 1 and coupled at least one stylobate are because of sequenator.Particular content repeats no more with reference to the statement in the earlier figures 1 here.This method flow comprises the steps: that system 1 controlling gene sequenator imports reaction system with reagent, and the temperature of conditioned reaction system; System 1 control reaction system moving in gene sequencer, and confirm to adopt the figure position; The affinity tag that Nucleotide carries in the system 1 provocative reaction system is luminous, and obtains picture signal adopting the figure position.
Fig. 7 shows in the one embodiment of the invention method flow that robotization is controlled is carried out in biological sample processing and order-checking, and this method flow is based on aforementioned system shown in Figure 2.This system comprises system 1 and coupled at least one stylobate because of sequenator, and particular content repeats no more here.
Method flow shown in Figure 7 may further comprise the steps:
Step S0, the preparation of control biological sample, the formation reaction system also is set to it in gene sequencer.About the particular content of step S1, will in embodiment thereafter, set forth in detail.
Step S1, system 1 utilizes its reaction control unit 100 controlling gene sequenators that reagent is imported reaction system, and the temperature of conditioned reaction system.About the particular content of step S1, will in embodiment thereafter, set forth in detail.
Step S2, system 1 is utilized its Positioning Control Unit 200 control reaction systems moving in gene sequencer, and confirms to adopt the figure position.About the particular content of step S2, will in embodiment thereafter, set forth in detail.
Step S3, it is luminous that system 1 utilizes it to adopt the affinity tag that Nucleotide carries in the figure control unit 300 provocative reaction systems, and obtain picture signal adopting the figure position.About the particular content of step S3, will in embodiment thereafter, set forth in detail.
In concrete the application, aforesaid method shown in Figure 7 carries out automated operation respectively to each corresponding in gene sequencer assembly, and can effectively coordinate different assemblies.What is more important; Every operation has all taken into full account the technical characterstic in each stage of gene sequencing; Comprise the control of step S1 to reagent dosage and type, temperature of reaction, time, cleanliness factor etc.; Step S2 is to the control of nano-grade displacement, focus adjustment etc., and step S3 all operates according to strict index the control of luminous intensity, light path adjusting, time shutter calculating, image taking etc. accurately.Therefore aforesaid method can increase substantially the stability and the efficient of order-checking process, and the accuracy of sequencing result.
Fig. 8 shows step S0 method flow in one embodiment among Fig. 7, comprising:
Step S01 prepares gene fragment storehouse to be measured according to biological sample, and generates the required reaction system of order-checking.In the present invention, the specimen preparation process of reaction system realizes through a water in oil unique DNA fragment amplification system.In water in oil system, comprise a large amount of separate reactions and drip, each reaction is dripped and is comprised a microballon, is combined with dna fragmentation to be measured on it.Through water in oil system is carried out pcr amplification, each magnetic bead will combine a plurality of copy number target DNA fragments after the amplification, and these fragments all come from same dna profiling to be measured.Certainly, the present invention also can also can carry out subsequent operations to unit molecule without pcr amplification, just behind pcr amplification, can strengthen the order-checking signal, improves sequencing quality.
In one embodiment; Situation with the gene sequencer assembly described among Fig. 2 is an example; Step S01 specifically comprises: (a) system prepares module 401 transmission instructions and prepares assembly to the library; Control this assembly and under different experiment conditions, biological sample is carried out a series of processing, make biological sample accomplish the transformation of several stages, obtain gene fragment storehouse to be measured.What need explanation is that (b) system prepares module 401 transmission instructions and prepares assembly to reaction system, controls the mixing that this assembly carries out oil phase, water, generates water in oil reaction system under certain condition.
In above-mentioned (a), as if the type of the biological sample that obtains at first, stage of living in difference, system prepares module 401 will corresponding multiple different processing mode.In one embodiment, if the biological sample that obtains at first is histocyte, bacterium etc., system prepares module 401 and will at first control the library and prepare assembly and from these histocytes, bacterium, extract RNA so, and transcribes the synthetic initial dna fragmentation that obtains.Then initial dna fragmentation is attached on the microballon; To be combined in a plurality of initial dna fragmentation enzyme on the microballon again and be cut into a plurality of DNA labels of equal in length; Then each DNA label is connected one section universal sequence; At last each the DNA label that is connected with universal sequence is increased, obtain gene fragment storehouse to be measured.In another embodiment; If the biological sample that obtains at first has been through extracting, transcribe the synthetic initial dna fragmentation that obtains; System prepares 401 of modules control library and prepares assembly initial dna fragmentation is attached on the microballon so, will be combined in a plurality of DNA labels that a plurality of initial dna fragmentation enzyme on the microballon is cut into equal in length again, then each DNA label is connected one section universal sequence; At last each the DNA label that is connected with universal sequence is increased, obtain gene fragment storehouse to be measured.
In above-mentioned (b), the preparation of water-in-oil reaction system has multiple mode.In one embodiment, system prepares module 401 at first to be controlled reaction system and prepares assembly and set up the microballon comprise after the modification at interior PCRmix aqueous phase system, for example sets up following aqueous phase system: (1) 10 * PCR buffer, 15 μ L; (2) dNTP, 3 μ L; (3) dna profiling, 2 μ L; (4) microballon after primer combines, 5 μ L; (5) Taq enzyme, 9 μ L; (6) Mg2+, 3 μ L; (7) ddH
2O, 114 μ L; (8) preferred adding can add a pair of little primer of fast response toggle speed, each 0.75 μ L of two ends, upstream and downstream in aqueous phase system.System prepares module 401 to be controlled reaction system then and prepares assembly and set up the oil phase system that oils solution constitutes, and then aqueous phase system is injected into the oil phase system, and shaking mixing under certain condition becomes emulsion.This condition is for example shaken frequency 15HZ, time 15s.Above-mentioned water-in-oil reaction system can prepare in multiple container, for example in the test tube.
(2) going up appearance control module 402 is set to reaction system in the gene sequencer.
Situation with the gene sequencer assembly described among Fig. 2 is an example, and this setting up procedure is to utilize the mechanical manipulator of going up in 402 pairs of gene sequencers of appearance control module to control, and reaction system is set in the movable assembly in the gene sequencer.In one embodiment, last appearance control module 402 is sent instruction and is given mechanical manipulator, and the water-in-oil reaction system in the aforesaid test tube is imported a reaction small chamber, reaction small chamber is installed to the specific position of sample table again.
So far, accomplished the specimen preparation of robotization and gone up the appearance operation, then can begin to check order.
Fig. 9 shows step S1 method flow in one embodiment among Fig. 7, and this method flow is based on Fig. 1, system shown in Figure 2.In this system, reaction control unit 100 comprises reagent control module 101, temperature control module 102.Step S1 comprises:
Step S11, reagent control module 101 controlling gene sequenators are selected reagent, and draw corresponding reagent, import reaction system.There is multiple concrete implementation in step S11 among the present invention; If be the situation of description in the earlier figures 2 in the gene sequencer with reaction control unit 100 corresponding assemblies; Then its control process is: reagent control module 101 is confirmed the required types of agents of taking of different steps; Send instructions to mechanical manipulator, the control mechanical manipulator moves to reagent position corresponding on the reagent platform, and the flexible pipe that will be fixed on the mechanical manipulator inserts in the reagent; Reagent control module 101 sends instructions to pump, thereby reagent is drawn in the control pump running; Reagent control module 101 sends instructions to pump after being drawn to required reagent, continues the control pump running reagent is squeezed into reaction small chamber.
Step S12, temperature control module 102 is controlled at the required temperature of reaction with the temperature of reaction system.There is multiple concrete implementation in step S12 among the present invention, will set forth in detail through various embodiment below.
In one embodiment; Situation with the gene sequencer assembly described among Fig. 2 is an example; Step S12 realizes through the serial communication mode; Detailed process is: temperature control module 102 sends instruction and gives TP, and the controlled temperature transmitter detects the temperature of reaction small chamber, and reads temperature detection result t; Be provided with the temperature value T in differential responses stage in the temperature control module 102, after it obtains temperature detection result t, then it compared with the temperature value T that is provided with; Temperature control module 102 is further handled according to comparing result; If t＜T, temperature control module 102 sends instruction and gives the heat riser in the temperature controller, and the heat riser in the control temperature controller starts; Heat to reaction small chamber; If t >=T then need not start the heat riser heating, heat riser is cooled to temperature T automatically through outside atmosphere.
For the gene sequencer assembly under other situations, control principle is consistent, and detailed process possibly there are differences.For example; In another embodiment; If the contrast of describing among gene sequencer assembly and Fig. 2, temperature controller except comprise the TP that is used for the little chambers temp of detection reaction, to the heat riser that reaction small chamber heats, also comprise to reaction small chamber refrigerating heat sink.So in this case, the implementation procedure of step S12 is: temperature control module 102 sends instruction and gives TP, and the controlled temperature transmitter detects the temperature of reaction small chamber, and reads temperature detection result t; Be provided with the temperature value T in differential responses stage in the temperature control module 102, after it obtains temperature detection result t, then it compared with the temperature value T that is provided with; Temperature control module 102 is further handled according to comparing result, if t＜T, temperature control module 102 sends instruction and gives the heat riser in the temperature controller; Heat riser in the control temperature controller starts; To the reaction small chamber heating, if t >=T, temperature control module 102 sends instruction and gives the heat sink in the temperature controller; Heat sink in the control temperature controller starts, and reaction small chamber is freezed.
Can know by Fig. 9; Step S11 can be to types of agents, reagent dosage, capture velocity, get speed etc. controls accurately; Step S12 can carry out strictness control to temperature detection, temperature setting, heating, refrigeration etc.; Thereby the biochemical reaction process that has guaranteed reaction system carries out smoothly, has also therefore improved stability, efficient and the accuracy of whole measuring program process.
Figure 10 shows step S2 method flow in one embodiment among Fig. 7.
Step S21, the current position of detection reaction system in gene sequencer, and control it and move to its target location of place on plane.Among the present invention, step S21 can control moving of reaction system in several ways.
In one embodiment; Be the situation of describing in the earlier figures 2 with Positioning Control Unit 200 corresponding assemblies in the gene sequencer; Step S21 realizes through the serial communication mode: displacement module 201 at first sends instruction and gives sample table; Reading the starting position coordinate of sample table on its plane, place, for example is (X
0, Y
0); Confirm reaction system at its place the target coordinate on plane (X, Y) after, sample table is given in displacement module 201 instruction of redispatching, the control sample table is from (X
0, Y
0) move to target coordinate (X, Y).
Step S22, the focus adjustment of controlling gene sequenator, that confirms reaction system adopts the figure position.Among the present invention, what step S22 can confirm reaction system in several ways adopts the figure position, and the situation with the gene sequencer assembly described among Fig. 2 is that example describes below.
In one embodiment; Step S22 realizes through the serial communication mode: focus module 202 is at first sent instruction and is given microscope; The control microscope with the sample table vertical direction on move; Through regulating the distance between microscope and the reaction small chamber, the position that sharpness is best is confirmed as and is adopted the figure position.
In another embodiment; Step S22 remains and realizes through the serial communication mode: focus module 202 is sent instruction and is given sample table; The control sample table moves on its place planar vertical direction; Through the distance between conditioned reaction cell and the microscope, the position that sharpness is best is confirmed as and is adopted the figure position.
In above-mentioned two embodiment, step S22 can confirm the position that image definition is best in several ways.For example, can observe the position that mode of comparing confirms that image definition is best, perhaps utilize the arithmetic calculation image definition, utilize algorithm to regulate image definition automatically through the microscope camera lens, or the like.
By on can know; Step S21 can carry out accurate control automatically to the displacement of micro/nano level; Step S22 can carry out accurate control automatically to focus adjustment, definition judgment etc.; Thereby can confirm the best figure position of adopting quickly and accurately, also therefore improve stability, efficient and the accuracy of order-checking process.
Figure 11 shows step S3 method flow in one embodiment among Fig. 7.
Step S31, the excitation light irradiation reaction system of control specific wavelength makes the affinity tag that Nucleotide carries in the reaction system luminous.In the present invention, step S31 can be accomplished in several ways.
In one embodiment; Be the situation of describing in the earlier figures 2 with adopting figure control unit 300 corresponding assemblies in the gene sequencer; The implementation procedure of step S31 is so: excitation module 301 is sent instruction and is given excitation light source; It is luminous to start excitation light source, makes the reaction small chamber of irradiate light on the sample table.In the present embodiment, the affinity tag that Nucleotide carries on the microballon in the reaction system is a fluorescent marker, and it just can send fluorescence after receiving the light source activation of specific wavelength.
Step S32 confirms the time shutter, and adopts and should the time shutter reaction system be taken pictures, and obtains picture signal.In the present invention, step S32 can be accomplished in several ways.
Continuation is in the embodiment of abovementioned steps S31; Be the situation of describing in the earlier figures 2 with adopting figure control unit 300 corresponding assemblies in the gene sequencer; So in step S32; At first confirm suitable exposure time values, send instruction then and give CCD that control CCD takes fluorogram according to this exposure time values by photo module 302.
Step S32 of the present invention can confirm for example according to circumstances artificially to be provided with suitable exposure time values in several ways, and the process cumulative time shutter empirical value that perhaps is set to repeatedly check order perhaps goes out suitable exposure time values through arithmetic calculation, or the like.In the gene sequencing system of prior art before this, major part has adopted the mode of artificial setting, and the present invention then has the plurality of optional pattern, is intended to confirm the best exposure time value according to condition of different, thereby improves the quality of picture signal.
Step S33 preserves the picture signal of obtaining.Image access module 303 can adopt multiple format picture signal among the present invention.Step S33 can be accomplished in several ways.
Continuation is in the embodiment of abovementioned steps S31, S32; The implementation of step S33 is: photo module 302 sends to image access module 303 with the fluorogram that CCD takes; Image access module 303 can adopt special high fidelity visual storage format to preserve fluorogram; Also can adopt common image storage format, for example TIFF, EPS, PNG, PSD form etc.
By on can know; Step S31 can accurately control luminous light path of excitation light source etc.; Step S32 can accurately control the confirming of exposure time values, image taking etc., and step S33 can adopt best picture format memory image signal, thus the quality of the picture signal that has guaranteed to be obtained; Greatly improved the accuracy of sequencing result, and this robotization mode the stability and the efficient of order-checking process have also been improved.
In order more clearly to explain the present invention, the applicant will be the whole process that gene sequencing was handled and carried out to known biological sample of example explanation with a concrete experimentation.This application scenarios is to adopt certain known sequence measurement that DNA is checked order:
(1) from the bacteria samples of being obtained, extracts RNA, transcribe the synthetic initial dna fragmentation that obtains.
(2) initial dna fragmentation is attached on the microballon, will be combined in a plurality of DNA labels that a plurality of initial dna fragmentation enzyme on the microballon is cut into equal in length again, then each DNA label is connected one section universal sequence.
(3) each the DNA label that is connected with universal sequence is increased, obtain gene fragment storehouse to be measured.
(4) 3 ' the terminal modified microballon on the testing gene fragment is deposited on the appearance slide, in deposition process, can regulates, to reach flux peak to microballon density.
(5) eight polynucleotides that in reaction system, add dna ligase, universal sequencing primer thing n and have 3 '-XXnnnzzz-5 ' structure.In this eight polynucleotide, the base on the 1st and the 2nd (XX) is confirmed, and has been added different fluorescent marks on the 6-8 position (zzz) that do not coexist according to kind.This sequence measurement by two base decisions is called as two bases order-checkings (two base encoding).
(6) when eight polynucleotides owing to the 1st be connected last time by ligase enzyme with the 2nd bit pairing, through the optical excitation of specific wavelength, can send fluorescence.
(7) after noting fluorescence information, cut between the 5th and the 6th through chemical process, the cancellation fluorescent signal is to carry out the order-checking of next position.
By this method, the position of each order-checking all differs five, promptly for the first time surveys the 1st and the 2nd, surveys the 6th and the 7th for the second time ... after measuring the end, with the sex change of new synthetic chain, wash-out.Then carry out second and take turns order-checking with universal sequencing primer thing n-1.The difference of universal sequencing primer thing n-1 and universal sequencing primer thing n is, the two with joint paired position on differ a base, promptly universal sequencing primer thing n-1 has moved a base to 3 ' end on universal sequencing primer thing n mated position.Therefore after adding dna ligase and eight polynucleotides; Can measure the 0th and the 1st, the 5th and the 6th ... second takes turns order-checking accomplish after; Next add respectively again that universal sequencing primer thing n-2, universal sequencing primer thing n-3, universal sequencing primer thing n-4 carry out third round, four-wheel, the 5th is taken turns order-checking, finally can accomplish the mensuration of whole positions.
Above-mentioned order-checking process, each take turns relate to all repeatedly that reagent is taken, temperature adjusting, time control etc., no matter be simple manual operation, or the MC instrumentation, all can't fully guarantee stability, efficient and the accuracy of testing.And utilize control method of the present invention and system, and only need to be arranged on the reaction system for preparing in the gene sequencer, select order-checking pattern to different samples, just can make gene sequencer above-mentioned each step of operation automatically, need not manual operations.If based on system shown in Figure 2 and method shown in Figure 7, through the automatic sequencing process of gene sequencer, can collect picture signal fast after the last appearance; If based on system shown in Figure 3 and method shown in Figure 8, through the automatic sequencing process of gene sequencer, can directly obtain gene order information after last kind, for example base puts in order, Disease-causing gene site etc.
Should be noted that method and system of the present invention are applicable to handles various types of biological samples, and on this basis the order-checking process of gene sequencer is carried out robotization control.Even if dissimilar biological samples possibly there are differences on processing mode; Perhaps dissimilar gene sequencers possibly there are differences on concrete internal structure; But above-mentioned system and control method are consistent or similar on grammar; Therefore protection scope of the present invention should not receive dissimilar biological samples, the restriction of the internal structure of perhaps dissimilar gene sequencers.
The above is merely preferred embodiment of the present invention, not in order to restriction the present invention, all any modifications of within spirit of the present invention and principle, being done, is equal to and replaces and improvement etc., all should be included within protection scope of the present invention.
Claims (12)
1. biological sample handled and system that robotization control is carried out in order-checking for one kind, it is characterized in that said system comprises sample control unit, reaction control unit, Positioning Control Unit, adopts the figure control unit;
Said sample control unit is used to control the preparation of biological sample, and the formation reaction system also is set to it in gene sequencer;
Said reaction control unit is used for the controlling gene sequenator reagent is imported reaction system, and in the order-checking process temperature of conditioned reaction system;
Said Positioning Control Unit is used for controlling reaction system the moving of gene sequencer, and confirms to adopt the figure position;
It is luminous that the said figure of adopting control unit is used for the affinity tag that provocative reaction system Nucleotide carries, and obtain picture signal in the said figure of adopting position.
2. according to claim 1ly the system that biological sample is handled and robotization control is carried out in order-checking is characterized in that said sample control unit comprises that system prepares module, goes up an appearance control module;
Said system prepares module and prepares gene fragment storehouse to be measured according to biological sample, and is processed into the required reaction system of order-checking;
The said appearance control module that goes up is set in the gene sequencer allocation really with reaction system.
3. according to claim 1 to the system that biological sample is handled and robotization control is carried out in order-checking, it is characterized in that said reaction control unit comprises reagent control module, temperature control module;
Said reagent control module is used for the controlling gene sequenator to be selected reagent, and draws corresponding reagent, imports reaction system;
Said temperature control module is used for the temperature of reaction system is controlled at the required temperature of reaction.
4. according to claim 1 to the system that biological sample is handled and robotization control is carried out in order-checking, it is characterized in that said Positioning Control Unit comprises displacement module, focus module;
Said displacement module is used for the current position of detection reaction system at gene sequencer, and controls it and move to its target location of place on plane;
Said focus module is used for the focus adjustment of controlling gene sequenator, and that confirms reaction system adopts the figure position.
5. according to claim 4ly the system that biological sample is handled and robotization control is carried out in order-checking is characterized in that said focus module is through regulating the distance between microscope and the reaction small chamber, the position that sharpness is best is confirmed as and is adopted the figure position.
6. according to claim 1 to the system that biological sample is handled and robotization control is carried out in order-checking, it is characterized in that the said figure of adopting control unit comprises excitation module, photo module, image access module;
Said excitation module is used to control the excitation light irradiation reaction system of specific wavelength, makes the affinity tag that Nucleotide carries in the reaction system luminous;
Said photo module is confirmed the time shutter, and adopts the said time shutter that reaction system is taken pictures, and obtains picture signal;
Said image access module and photo module communicate, and are used to preserve the picture signal of obtaining.
7. one kind according to the method that the said system of claim 1 handles biological sample and robotization control is carried out in order-checking, it is characterized in that, said method comprising the steps of:
A. control the preparation of biological sample, the formation reaction system also is set to it in gene sequencer;
B. the controlling gene sequenator imports reaction system with reagent, and the temperature of conditioned reaction system;
C. control reaction system moving in gene sequencer, and confirm to adopt the figure position;
D. the affinity tag that Nucleotide carries in the provocative reaction system is luminous, and obtains picture signal in the said figure of adopting position.
8. according to claim 7 to the method that biological sample is handled and robotization control is carried out in order-checking, it is characterized in that said steps A comprises:
A1. prepare gene fragment storehouse to be measured according to biological sample, and be processed into the required reaction system of order-checking;
A2. said reaction system is set in the gene sequencer allocation really.
9. according to claim 7 to the method that biological sample is handled and robotization control is carried out in order-checking, it is characterized in that said step B comprises:
B 1. controlling gene sequenators are selected reagent, and draw corresponding reagent, import reaction system;
B2. the temperature with reaction system is controlled at the required temperature of reaction.
10. according to claim 7 to the method that biological sample is handled and robotization control is carried out in order-checking, it is characterized in that said step C comprises:
C1. the current position of detection reaction system in gene sequencer, and control it and move to the target location on its plane, place;
C2. the focus adjustment of controlling gene sequenator, that confirms reaction system adopts the figure position.
11. according to claim 10 to the method that biological sample is handled and robotization control is carried out in order-checking, it is characterized in that said step C2 comprises:
Through regulating the distance between microscope and the reaction small chamber, the position that sharpness is best is confirmed as and is adopted the figure position.
12. according to claim 7 to the method that biological sample is handled and robotization control is carried out in order-checking, it is characterized in that said step D comprises:
D1. control the excitation light irradiation reaction system of specific wavelength, make the affinity tag that Nucleotide carries in the reaction system luminous;
D2. confirm the time shutter, and adopt the said time shutter that reaction system is taken pictures, obtain picture signal;
D3. preserve the picture signal of obtaining.
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| WO2020037571A1 (en) * | 2018-08-22 | 2020-02-27 | 深圳市真迈生物科技有限公司 | Method and apparatus for building sequencing template on basis of images, and computer program product |
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