CN102146442B - Method and system for controlling sequencing process of gene sequencer - Google Patents
Method and system for controlling sequencing process of gene sequencer Download PDFInfo
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Abstract
The invention relates to the field of genetic engineering and provides a method and a system for controlling the sequencing process of a gene sequencer. The system comprises a reaction control unit, a positioning control unit and an image acquisition control unit, wherein a DNA fragment sample to be tested is arranged in the reaction small chamber of the gene sequencer. The method comprises the following steps: A, controlling the gene sequencer to introduce the reagent into the reaction small chamber, and regulating the temperature of the reaction small chamber; B, controlling the movement of the reaction small chamber in the gene sequencer, and determining the image acquisition position of the DNA fragment sample to be tested; and C, exciting a maker carried by the nucleotide of the DNA fragment sample to be detected, and acquiring an image signal at the image acquisition position. In the invention, through the automatic control over the sequencing process of the gene sequencer, the stability and efficiency of the sequencing process are improved, and the accuracy of the sequencing result is improved. In addition, sufficient sequencing flux is ensured by controlling the gene sequencer to perform large-scale image acquisition.
Description
Technical field
The present invention relates to genetically engineered field, more particularly, relate to a kind of method and system that the order-checking process of gene sequencer is controlled.
Background technology
Initial gene sequencing technology is undertaken by manual operations, comprises the dideoxy chain termination of Sanger invention, and the chemical degradation method of Maxam and Gilbert invention.Because efficiency hand-manipulated is lower, and human operational error easily occurs, therefore utilizing gene sequencer to check order has now become the main flow of sequencing technologies.
Current gene sequencer, its order-checking process is made up of operations such as a series of machinery, electronic communication, biology, chemistry, optics, and these operations are performed by assembly corresponding in gene sequencer respectively, substituted simple manual operations.But gene sequencer also faces 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 occurs that deviation all can cause sequencing result undesirable; On the other hand, the concrete steps that whole order-checking process relates to are very loaded down with trivial details, need to carry out Collaboration between the each assembly in gene sequencer.That is to say, order-checking process not only requires each assembly in gene sequencer to carry out accurately and rapidly operations, also requires to carry out between each assembly good cooperation.In concrete application, the factor relating 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, optical path adjusting, time shutter calculating, image takings, and the requirement of each aspect is very high, therefore will ensure that order-checking process carries out smoothly, difficulty is very large.
Only describe with the control of reagent dosage and type, due in gene sequencing process to the control of reagent dosage generally at micro updating, and need to carry out repeatedly in different step of reaction the absorption importing of various dose, add that each selected types of agents all may there are differences, therefore the assurance of the dosage to reagent, type has proposed higher requirement.If carry out reagent absorption by manual operation, or Artificial Control instrument carries out reagent absorption, all has following problem: be difficult on the one hand accurately control dosage, and the nuance of dosage can cause different biochemical reaction results, also will directly affect sequencing result; On the other hand, artificial participation need to accurately judge all ingredients type of reaction different steps, even if the error in an annelet will cause biochemical reaction failure, makes the failure totally of whole order-checking process.In addition, because order-checking process is from sample preparation, loading, order-checking, data analysis until draw sequencing result, each stage needs certain cycle, if there is error in the control of mentioned reagent dosage and type, manual operation cannot be monitored, can not be in subsequent process error correction in time, even if learn that final sequencing result failure is also difficult to find the basic reason of order-checking procedure failure, has also wasted a large amount of time and expensive reagent.
Except the factor of reagent dosage and type, other various factorss, comprise aforesaid temperature of reaction, time, cleanliness factor, nano-grade displacement, focus adjustment, luminous intensity, optical path adjusting, time shutter calculating, image taking etc., all there is above-mentioned similar situation, if do not have the Controlling System of automatization to operate, whole order-checking process will be difficult to launch smoothly, and want to stablize, obtain rapidly, sequencing result is just more difficult accurately.In addition, utilize general order-checking mode to carry out gene sequencing, can not adopt on a large scale figure, flux is lower.
Therefore need a kind of method and system of the order-checking process of gene sequencer being carried out to automatization control, can realize the automatization control to order-checking process, reduce manual operations.
Summary of the invention
The object of the present invention is to provide a kind of method and system that the order-checking process of gene sequencer is controlled, be intended to realize the automatization control to order-checking process, reduce manual operations.
In order to realize goal of the invention, the invention provides a kind of system that the order-checking process of gene sequencer is controlled, wherein DNA fragmentation sample to be measured is arranged in the reaction small chamber of gene sequencer, and described system comprises reaction control unit, Positioning Control Unit, adopts figure control unit; Described reaction control unit imports reaction small chamber for controlling gene sequenator by reagent, and in order-checking process, regulates the temperature of reaction small chamber; Described Positioning Control Unit is for controlling the movement of reaction small chamber at gene sequencer, and in definite reaction small chamber DNA fragmentation sample to be measured adopt figure position; The described figure of adopting control unit is luminous for the marker that excites DNA fragmentation sample Nucleotide to be measured to carry, and in the described figure position acquisition picture signal of adopting.
Wherein, adopting in figure process, described system controlling gene sequenator circulates and adopts figure the position one by one on DNA fragmentation sample to be measured in reaction small chamber; Described Positioning Control Unit control reaction small chamber successively moves in gene sequencer, and determine each mobile after in reaction small chamber DNA fragmentation sample to be measured adopt figure position; The described figure of adopting control unit excites the marker that in DNA fragmentation sample to be measured, Nucleotide carries luminous, and after described each movement in reaction small chamber DNA fragmentation sample to be measured adopt figure position acquisition picture signal.
Wherein, described reaction control unit comprises reagent control module, temperature control module; Described reagent control module is selected reagent for controlling gene sequenator, and draws corresponding reagent, imports reaction small chamber; Described temperature control module is for being controlled at the temperature of reaction small chamber the required temperature of reaction.
Wherein, described Positioning Control Unit comprises displacement module, focus module; Described displacement module is the current position in gene sequencer for detection of reaction small chamber, and control its move to its target location in the plane; Described focus module is for the focus adjustment of controlling gene sequenator, and that determines DNA fragmentation sample to be measured in reaction small chamber adopts figure position.
Wherein, described focus module is by regulating the distance between microscope and reaction small chamber, by the location positioning of sharpness the best for adopting figure position.
Wherein, described in, adopt figure control unit and comprise excitation module, photo module, image access module; Described excitation module, for controlling the excitation light irradiation reaction small chamber DNA fragmentation sample to be measured of specific wavelength, makes the marker that in DNA fragmentation sample to be measured, Nucleotide carries luminous; Described photo module is determined the time shutter, and adopts the described time shutter to take pictures to DNA fragmentation sample to be measured in reaction small chamber, obtains picture signal; Described image access module and photo module communicate, for preserving the picture signal of obtaining.
In order to realize better goal of the invention, the present invention also provides a kind of method of the order-checking process of gene sequencer being controlled based on said system, wherein DNA fragmentation sample to be measured is arranged in the reaction small chamber of gene sequencer, said method comprising the steps of: reagent is imported reaction small chamber by A. controlling gene sequenator, and regulate the temperature of reaction small chamber; B. control the movement of reaction small chamber in gene sequencer, and in definite reaction small chamber DNA fragmentation sample to be measured adopt figure position; C. excite the marker that in DNA fragmentation sample to be measured, Nucleotide carries luminous, and in the described figure position acquisition picture signal of adopting.
Wherein, adopting in figure process, circulated and adopt figure in position one by one on DNA fragmentation sample to be measured in reaction small chamber, comprise the following steps: B '. control reaction small chamber and successively move in gene sequencer, and determine each mobile after in reaction small chamber DNA fragmentation sample to be measured adopt figure position; C '. excite the marker that in DNA fragmentation sample to be measured, Nucleotide carries luminous, and after described each movement in reaction small chamber DNA fragmentation sample to be measured adopt figure position acquisition picture signal.
Wherein, described steps A comprises: A1. controlling gene sequenator is selected reagent, and draws corresponding reagent, imports reaction small chamber; A2. the temperature of reaction small chamber is controlled to the required temperature of reaction.
Wherein, described step B or B ' comprising: the current position of B1. detection reaction cell in gene sequencer, and control its move to its target location in the plane; B2. the focus adjustment of controlling gene sequenator, that determines DNA fragmentation sample to be measured in reaction small chamber adopts figure position.
Wherein, described step B2 comprises: by regulating the distance between microscope and reaction small chamber, by the location positioning of sharpness the best for adopting figure position.
Wherein, described step C or C ' comprising: DNA fragmentation sample to be measured in the excitation light irradiation reaction small chamber of C1. control specific wavelength, makes the marker that in DNA fragmentation sample to be measured, Nucleotide carries luminous; C2. determine the time shutter, and adopt the described time shutter to take pictures to DNA fragmentation sample to be measured in reaction small chamber, obtain picture signal; C3. preserve the picture signal of obtaining.
As from the foregoing, the method and system that the order-checking process of gene sequencer is controlled of the present invention, realize the automatization control to order-checking process, reduce manual operations, and can further realize the monitoring to operating process.
Brief description of the drawings
Fig. 1 is the system architecture schematic diagram that the present invention controls the order-checking process of gene sequencer;
Fig. 2 is Controlling System 1 in Fig. 1 structural representation in one embodiment;
Fig. 3 is reaction control unit 100 in Fig. 2 structural representation in one embodiment;
Fig. 4 is Positioning Control Unit 200 in Fig. 2 structural representation in one embodiment;
Fig. 5 adopts figure control unit 300 structural representation in one embodiment in Fig. 2;
Fig. 6 is the method flow diagram that in one embodiment of the invention, the order-checking process to gene sequencer is controlled;
Fig. 7 is step S1 method flow diagram in one embodiment in Fig. 6;
Fig. 8 is step S2 method flow diagram in one embodiment in Fig. 6;
Fig. 9 is step S3 method flow diagram in one embodiment in Fig. 6;
Figure 10 is the schematic diagram that utilizes system and method for the present invention to check order to DNA.
Embodiment
In order to make object of the present invention, technical scheme and advantage clearer, below in conjunction with drawings and Examples, the present invention is further elaborated.
Fig. 1 shows the system architecture that the present invention controls the order-checking process of gene sequencer, and this system comprises Controlling System 1, and at least one coupled gene sequencer, gene sequencer 2 as shown in the figure, gene sequencer 3 ... gene sequencer N.It should be noted that in all diagrams of the present invention, the annexation between each equipment is for the needs of clear its information interaction of explaination and control process, therefore should be considered as control relation in logic, and should not be limited to physical connection or wireless connections.It should be noted that in addition, the signalling methods between each functional module can be taked multiple, and protection scope of the present invention should not be defined as the signalling methods of certain particular type.Wherein:
(1) Controlling System 1 is for communicating with at least one gene sequencer, and its each functional module is distinguished each assembly corresponding in controlling gene sequenator, thus the order-checking process of controlling gene sequenator.Mainly comprise: reagent is imported reaction small chamber by controlling gene sequenator, and regulate the temperature of reaction small chamber; Control the movement of reaction small chamber in gene sequencer, and in definite reaction small chamber DNA fragmentation sample to be measured adopt figure position; Excite the marker that in DNA fragmentation sample to be measured, Nucleotide carries luminous, and in the above-mentioned figure position acquisition picture signal of adopting.It should be noted that above-mentioned control mode is applicable to various types of gene sequencers, therefore in the present invention, the protection domain of control method and system should not be subject to the restriction of this body structure of gene sequencer.About the particular content of Controlling System 1, will in embodiment thereafter, elaborate.
(2) gene sequencer N is by multiple module compositions, corresponding with each functional module in Controlling System 1 respectively, accepts and carries out the instructions of these functional modules, thereby worked in coordination with order-checking.These assemblies comprise: for drawing reagent and importing the assembly of reaction small chamber, for the assembly that the temperature of reaction small chamber is regulated, the assembly that comprises reaction small chamber and can move in gene sequencer, for determining the assembly of adopting figure position of reaction small chamber DNA fragmentation sample to be measured, for importing the assembly of exciting light, for gathering the assembly etc. of picture signal.It should be noted that dissimilar gene sequencer has different intrawares, or the external expressive form of intraware is different, but the function realizing is consistent, protection scope of the present invention should not be subject to the restriction of these factors.Also it should be noted that between each assembly not necessarily completely independently, each assembly of realizing difference in functionality may relate to one or more identical parts.About the part module composition in gene sequencer N, publication that can application reference people: application number is CN200810132008.8, denomination of invention is " sequencing reaction small chamber, gene sequencing reaction bench and gene sequencing system ", and the present invention also will be specifically addressed in embodiment thereafter.
It should be noted that, before checking order, DNA fragmentation sample to be measured prepares, and is aligned in reaction small chamber, and this reaction small chamber is arranged in the sample table of gene sequencer.The preparation process of DNA fragmentation sample to be measured is: first extracts DNA from tissue, blood, bacterium etc., is processed into DNA fragmentation to be measured equal in length, and jointing sequence; Then by joint sequence, the primer on microballon is combined, and DNA fragmentation to be measured is attached on microballon; Be prepared into water in oil unique DNA fragment amplification system again, make to comprise a large amount of separate reactions in this system and drip, each reaction is dripped and is comprised a microballon that is combined with DNA fragmentation to be measured; Then this water in oil system is carried out to pcr amplification, make each microballon in conjunction with multiple copy number target DNA fragments, and these fragments all come from same DNA profiling to be measured; Microballon in again reaction being dripped takes out, enrichment, and point sample is aligned in reaction small chamber, finally reaction small chamber is installed in the sample table of gene sequencer.
Fig. 2 shows Controlling System 1 in Fig. 1 structure in one embodiment, comprises reaction control unit 100, Positioning Control Unit 200, adopts figure control unit 300.Wherein:
(1) reaction control unit 100 imports reaction small chamber for controlling gene sequenator by reagent, and in order-checking process, regulates the temperature of reaction small chamber.
In the present invention, gene sequencer inside has the multiple assemblies corresponding with reaction control unit 100, reaction control unit 100 is actually by the operation of control section assembly, reagent is imported to reaction small chamber, and by controlling the operation of another part assembly, realize the adjusting to reaction small chamber temperature.Examination taking the gene sequencer of a type as example illustrates above-mentioned control process, in this concrete situation, in gene sequencer, comprise and following assembly corresponding to reaction control unit 100: (1) reagent platform, for placing or holding the multiple reagent for drawing; (2) mechanical manipulator, for selecting suitable reagent in different step of reaction; (3) pump and flexible pipe, for drawing selected reagent, wherein flexible pipe is fixed on mechanical manipulator; (4) sample table, comprises reaction small chamber, is arranged with DNA fragmentation sample to be measured on reaction small chamber; (5) temperature controller, is connected with reaction small chamber, and it comprises the temperature sensor for detection of reaction small chamber temperature, and the heating unit heating to reaction small chamber.In this case, the control process of reaction control unit 100 is: by controlling mechanical manipulator, pump and flexible pipe, reagent is imported to reaction small chamber, and by temperature sensor detected temperatures, and control heating unit and heat to reaction small chamber, thereby the temperature of adjusting reaction small chamber.Should be noted that; for dissimilar gene sequencer; type, structure or the quantity of its included assembly may there are differences, but the control process of reaction control unit 100 is consistent in ultimate principle, and therefore protection domain should not be subject to the restriction of above-mentioned factor.About specific functional modules and the control mode of reaction control unit 100, will in embodiment thereafter, elaborate.
(2) Positioning Control Unit 200 is for controlling the movement of reaction small chamber at gene sequencer, and in definite reaction small chamber DNA fragmentation sample to be measured adopt figure position.
In the present invention, gene sequencer inside has the multiple assemblies corresponding with Positioning Control Unit 200, Positioning Control Unit 200 is actually by the operation of control section assembly, thereby control the movement of reaction small chamber in gene sequencer, and by controlling the operation of another part assembly, thereby determine DNA fragmentation sample to be measured in reaction small chamber adopt figure position.Examination taking the gene sequencer of a type as example illustrates above-mentioned control process, in this concrete situation, in gene sequencer, comprise the following assembly corresponding with Positioning Control Unit 200: (1) sample table, it is the sample table that aforementioned content is mentioned, it comprises reaction small chamber, is arranged with DNA fragmentation sample to be measured, as previously mentioned on reaction small chamber, not necessarily completely independent between each assembly, each assembly of realizing difference in functionality may relate to one or more identical parts; (2) microscope, for regulating focusing on.In this case, the control process of Positioning Control Unit 200 is: by the movement of Quality control platform, make reaction small chamber move to position suitable in gene sequencer, and by controlling the distance between microscope and reaction small chamber, thereby determine the suitable figure position of adopting.About specific functional modules and the control mode of Positioning Control Unit 200, will in embodiment thereafter, elaborate.
(3) adopt figure control unit 300 luminous for the marker that excites DNA fragmentation sample Nucleotide to be measured to carry, and in the above-mentioned figure position acquisition picture signal of adopting.
In the present invention, gene sequencer inside has the multiple assemblies corresponding with adopting figure control unit 300, adopting figure control unit 300 is actually by the operation of control section assembly, thereby excite the marker that in DNA fragmentation sample to be measured, Nucleotide carries luminous, and by controlling the operation of another part assembly, thereby adopting figure position acquisition picture signal.Examination taking the gene sequencer of a type as example illustrates above-mentioned control process, in this concrete situation, in gene sequencer, comprise the following assembly corresponding with adopting figure control unit 300: (1) light-emitting device, be used for continuing to send exciting light, this device also comprises a light-blocking member that is arranged in light path, be typically a shutter or optical gate, for block the light of exciting light in the time not needing excitation light irradiation; (2) photographic means, for photographic images.In this case, the control process of adopting figure control unit 300 is: send exciting light by controlling light-emitting device, excite the marker that in DNA fragmentation sample to be measured, Nucleotide carries luminous, and by controlling CCD photographic images, thereby the picture signal of obtaining.About specific functional modules and the control mode of adopting figure control unit 300, will in embodiment thereafter, elaborate.
System of the present invention can be controlled gene sequencer and carry out large-scale IMAQ, has clear superiority than general small throughput order-checking mode.Controlling System 1 can be circulated and adopt figure the position one by one on DNA fragmentation sample to be measured in reaction small chamber by controlling gene sequenator, reaches this point.Be embodied in and adopt in figure process Positioning Control Unit 200 and adopt coordinating of figure control unit 300: Positioning Control Unit 200 is controlled reaction small chamber and successively moved in gene sequencer, for example move with the spacing of a microballon at every turn, and determine each mobile after in reaction small chamber DNA fragmentation sample to be measured adopt figure position; Adopt figure control unit 300 and excite the marker that in DNA fragmentation sample to be measured, Nucleotide carries luminous, and the figure position of adopting of DNA fragmentation sample to be measured obtains picture signal to DNA fragmentation sample to be measured in each mobile rear reaction small chamber.
In concrete application, said system utilization reaction control unit 100, the Positioning Control Unit 200 shown in Fig. 2, adopt figure control unit 300 each assembly corresponding in gene sequencer is carried out respectively to automated operation, 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 control unit 100 to reagent dosage and type, temperature of reaction, time, cleanliness factor etc., the control of Positioning Control Unit 200 to nano-grade displacement, focus adjustment etc., adopt the control of figure control unit 300 to luminous intensity, optical path adjusting, time shutter calculating, image taking etc., all operate accurately according to strict index.Therefore system of the present invention can increase substantially stability and the efficiency of order-checking process, and the accuracy of sequencing result.In addition carry out large-scale IMAQ by controlling gene sequenator, therefore ensured enough sequencing throughput.
Fig. 3 shows reaction control unit 100 in Fig. 2 structure in one embodiment, comprises reagent control module 101, temperature control module 102.Wherein:
(1) reagent control module 101 is selected reagent for controlling gene sequenator, and draws corresponding reagent, imports reaction small chamber.
Taking the situation of the gene sequencer assembly described in Fig. 2 as example, the concrete control process of reagent control module 101 realizes by serial communication mode, detailed process is: reagent control module 101 is determined the required types of agents of taking of different steps, send instructions to mechanical manipulator, control mechanical manipulator and move to reagent position corresponding on reagent platform, and the flexible pipe being fixed on mechanical manipulator is inserted in reagent; Reagent control module 101 sends instructions to pump, thereby reagent is drawn in control pump running; Reagent control module 101 is drawn to after required reagent, sends instructions to pump, continues control pump running reagent is squeezed into reaction small chamber.
(2) temperature control module 102 is for being controlled at the temperature of reaction small chamber the required temperature of reaction.
Taking the situation of the gene sequencer assembly described in Fig. 2 as example, the concrete control process of temperature control module 102 realizes by serial communication mode, detailed process is: temperature control module 102 sends instruction to temperature sensor, control temperature sensor the temperature of reaction small chamber is detected, and read temperature detection result t; In temperature control module 102, be provided with the temperature value T in differential responses stage, when it obtains after temperature detection result t, it contrasted with the temperature value T arranging; Temperature control module 102 is further processed according to comparing result, if t<T, temperature control module 102 sends instruction to the heat riser in temperature controller, the heat riser of controlling in temperature controller starts, heat to reaction small chamber, if t >=T, does not need to start heat riser heating, heat riser is cooled to temperature T automatically by outside atmosphere.
For the gene sequencer assembly under other situations, control principle is consistent, and detailed process may there are differences.For example, in gene sequencer assembly under another kind of situation, than the situation of describing in Fig. 2, temperature controller, except comprising for detection of the temperature sensor of reaction small chamber temperature, to the heat riser of reaction small chamber heating, also comprises the heat sink to reaction small chamber refrigeration.So in this case, the concrete control process of temperature control module 102 is: temperature control module 102 sends instruction to temperature sensor, controls temperature sensor the temperature of reaction small chamber is detected, and read temperature detection result t; In temperature control module 102, be provided with the temperature value T in differential responses stage, when it obtains after temperature detection result t, it contrasted with the temperature value T arranging; Temperature control module 102 is further processed according to comparing result, if t<T, temperature control module 102 sends instruction to the heat riser in temperature controller, the heat riser of controlling in temperature controller starts, to reaction small chamber heating, if t>T, temperature control module 102 sends instruction to the heat sink in temperature controller, the heat sink of controlling in temperature controller starts, and reaction small chamber is freezed.
As from the foregoing, reagent control module 101 can be controlled accurately to types of agents, reagent dosage, reagent transmission speed etc., temperature control module 102 can strictly be controlled temperature detection, temperature setting, heating, refrigeration etc., thereby ensure that the biochemical reaction process in reaction small chamber carries out smoothly, also therefore improved stability, efficiency and the accuracy of whole order-checking process.
Fig. 4 shows Positioning Control Unit 200 in Fig. 2 structure in one embodiment, comprises displacement module 201, focus module 202.Wherein:
(1) displacement module 201 for detection of reaction small chamber the current position in gene sequencer, and control its move to its target location in the plane.In the present invention, displacement module 201 can be controlled the movement of reaction small chamber in several ways.
In one embodiment, in gene sequencer, the assembly corresponding with positioning unit 200 is the situation of describing in earlier figures 2, the concrete control process of displacement module 201 realizes by serial communication mode: first displacement module 201 sends instruction to sample table, read sample table its starting position coordinate in the plane, be for example (X
0, Y
0); Determine reaction small chamber its after object coordinate (X, Y) in the plane, displacement module 201 sends instruction again to sample table, Quality control platform is from (X
0, Y
0) move to object coordinate (X, Y).
Adopting in figure process, for controlling gene sequenator carries out large-scale IMAQ, need to circulate and adopt figure the position one by one on DNA fragmentation sample to be measured in reaction small chamber, therefore need displacement module 201 in figure process, to control reaction small chamber and carry out displacement according to position one by one adopting.For example, starting position (X
0, Y
0), in reaction small chamber, on DNA fragmentation sample to be measured, the corresponding figure position of adopting has n capable, and every row has m to adopt figure position, and each spacing of adopting between figure position equates.The capable j of i coordinate representation of adopting figure position is (X so
ij, Y
ij), the coordinate of all figure of adopting position can be expressed as: (X
11, Y
11), (X
12, Y
12) ..., (X
ij, Y
ij) ... (X
nm, Y
nm), wherein i, j, n, m are positive integer.In the time adopting figure, first displacement module 201 sends instruction to sample table, read sample table its starting position coordinate (X in the plane
0, Y
0), then sending instruction to sample table, Quality control platform is from (X
0, Y
0) move to object coordinate (X
11, Y
11); At (X
11, Y
11) position focuses on, adopts after figure completes, displacement module 201 sends instruction to sample table, and Quality control platform is from (X
11, Y
11) move to object coordinate (X
12, Y
12); At (X
12, Y
12) position focuses on, adopts after figure completes, displacement module 201 sends instruction to sample table, and Quality control platform is from (X
12, Y
12) move to object coordinate (X
13, Y
13).So move in circles, by the movement of position one by one, realize large-scale IMAQ.
(2) focus module 202 is for the focus adjustment of controlling gene sequenator, and that determines DNA fragmentation sample to be measured in reaction small chamber adopts figure position.In the present invention, what focus module 202 can be determined DNA fragmentation sample to be measured in several ways adopts figure position, describes below as an example of the situation of the gene sequencer assembly described in Fig. 2 example.
In one embodiment, the concrete control process of focus module 202 realizes by serial communication mode: first focus module 202 sends instruction to microscope, controlling microscope is moving up with sample table Vertical Square, by regulating the distance between microscope and reaction small chamber, by the location positioning of sharpness the best for adopting figure position.
In another embodiment, focus module 202 sends instruction to sample table, and Quality control platform moves up at the Vertical Square of its place plane, by regulating the distance between reaction small chamber and microscope, by the location positioning of sharpness the best for adopting figure position.
In above-mentioned two embodiment, focus module 202 can be determined the position of image definition the best in several ways.For example, the mode that can observe contrast by microscope camera lens be determined the position of image definition the best, or utilizes algorithm computed image sharpness, utilizes algorithm automatically to regulate image definition, etc.
As from the foregoing, 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 determine quickly and accurately the best figure position of adopting, also therefore improve stability, efficiency and the accuracy of order-checking process.
Fig. 5 shows and adopts figure control unit 300 structure in one embodiment in Fig. 2, comprises excitation module 301, photo module 302, image access module 303.Wherein:
(1) excitation module 301 is for controlling the excitation light irradiation reaction small chamber DNA fragmentation sample to be measured of specific wavelength, and the marker that Nucleotide is wherein carried is luminous.In the present invention, excitation module 301 can make marker luminous in several ways.
In one embodiment, in gene sequencer, the assembly corresponding with adopting figure control unit 300 is the situation of describing in earlier figures 2, excitation module 301 sends instruction to light-emitting device so, opens shutter or optical gate, and the irradiate light that makes exciting light is to the reaction small chamber in sample table.In the present embodiment, the marker that in reaction small chamber, in DNA fragmentation sample to be measured, on microballon, Nucleotide carries is fluorescent marker, and it is subject to just can sending fluorescence after the light source activation of specific wavelength.
(2) photo module 302 is determined the time shutter, and adopts the described time shutter to take pictures to DNA fragmentation sample to be measured in reaction small chamber, obtains picture signal.In the present invention, photo module 302 can be obtained picture signal in several ways.
In one embodiment, in gene sequencer, the assembly corresponding with adopting figure control unit 300 is the situation of describing in earlier figures 2, first photo module 302 determines suitable exposure time values so, then sends instruction to CCD, controls CCD and takes fluorogram according to this exposure time values.
Photo module 302 of the present invention can be determined suitable exposure time values in several ways, for example according to circumstances artificially arrange, or be set to the time shutter empirical value of repeatedly order-checking process accumulation, or calculate suitable exposure time values by algorithm, etc.In the gene sequencing Controlling System of prior art before this, major part has adopted the mode of artificial setting, and the present invention has plurality of optional pattern, is intended to determine best exposure time values according to different situations, thereby improves the quality of picture signal.
(3) image access module 303 communicates with photo module 302, for preserving the picture signal of obtaining.In the present invention, image access module 303 can adopt multiple format memory image signal.In the aforementioned embodiment, photo module 302 is controlled CCD and is taken after 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, such as TIFF, EPS, PNG, PSD or binary format etc.Protection scope of the present invention should not be subject to the restriction of image storage format.
As from the foregoing, the light path of the exciting light that excitation module 301 can be sent light-emitting device etc. is accurately controlled, photo module 302 can accurately be controlled the determining of exposure time values, image taking etc., image access module 303 can adopt best picture format memory image signal, thereby ensure the quality of the picture signal of obtaining, greatly improved the accuracy of sequencing result, and this automatization control mode stability and the efficiency of order-checking process are also improved.
The system of the method flow that the present invention controls the order-checking process of gene sequencer based on shown in Fig. 1, this system comprises Controlling System 1, and at least one coupled gene sequencer.Particular content, with reference to the statement in earlier figures 1, repeats no more herein.The method flow process comprises the steps: that reagent is imported reaction small chamber by Controlling System 1 controlling gene sequenator, and regulates the temperature of reaction small chamber; Controlling System 1 is controlled the movement of reaction small chamber in gene sequencer, and in definite reaction small chamber DNA fragmentation sample to be measured adopt figure position; Controlling System 1 excites the marker that in DNA fragmentation sample to be measured, Nucleotide carries luminous, and is adopting figure position acquisition picture signal.
Fig. 6 shows the method flow that in one embodiment of the invention, the order-checking process to gene sequencer is controlled, the system of the method flow process based on shown in earlier figures 2.This system comprises Controlling System 1, and at least one coupled gene sequencer, and particular content repeats no more herein.
It should be noted that DNA fragmentation sample to be measured prepares, and is aligned in reaction small chamber before carrying out the method flow process, this reaction small chamber is arranged in the sample table of gene sequencer.The preparation process of DNA fragmentation sample to be measured is: first extracts DNA from tissue, blood, bacterium etc., is processed into DNA fragmentation to be measured equal in length, and jointing sequence; Then by joint sequence, the primer on microballon is combined, and DNA fragmentation to be measured is attached on microballon; Be prepared into water in oil unique DNA fragment amplification system again, make to comprise a large amount of separate reactions in this system and drip, each reaction is dripped and is comprised a microballon that is combined with DNA fragmentation to be measured; Then this water in oil system is carried out to pcr amplification, make each microballon in conjunction with multiple copy number target DNA fragments, and these fragments all come from same DNA profiling to be measured; Microballon in again reaction being dripped takes out, enrichment, and point sample is aligned in reaction small chamber, finally reaction small chamber is installed in the sample table of gene sequencer.
Method flow shown in Fig. 6 comprises the following steps:
Step S1, Controlling System 1 utilizes its reaction control unit 100 controlling gene sequenators that reagent is imported to reaction small chamber, and regulates the temperature of reaction small chamber.About the particular content of step S1, will in embodiment thereafter, elaborate.
Step S2, Controlling System 1 utilizes its Positioning Control Unit 200 to control the movement of reaction small chamber in gene sequencer, and in definite reaction small chamber DNA fragmentation sample to be measured adopt figure position.About the particular content of step S2, will in embodiment thereafter, elaborate.
Step S3, Controlling System 1 is utilized it to adopt figure control unit 300 to excite the marker that in DNA fragmentation sample to be measured, Nucleotide carries luminous, and is adopting figure position acquisition picture signal.About the particular content of step S3, will in embodiment thereafter, elaborate.
Utilize aforesaid method can carry out large-scale IMAQ, there is clear superiority than general small throughput order-checking mode.The present invention can, by being circulated and adopt figure in the position one by one on DNA fragmentation sample to be measured in reaction small chamber, reach this point.Specifically comprise: step S2, Positioning Control Unit 200 is controlled reaction small chamber and is successively moved in gene sequencer, for example move with the spacing of a microballon at every turn, and determine each mobile after in reaction small chamber DNA fragmentation sample to be measured adopt figure position; Step S3, adopts figure control unit 300 and excites the marker that in DNA fragmentation sample to be measured, Nucleotide carries luminous, and after described each movement in reaction small chamber the figure position of adopting of DNA fragmentation sample to be measured DNA fragmentation sample to be measured is obtained to picture signal.
In concrete application, the aforesaid method shown in Fig. 6 carries out respectively automated operation to each assembly corresponding in gene sequencer, 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., the control of step S2 to nano-grade displacement, focus adjustment etc., the control of step S3 to luminous intensity, optical path adjusting, time shutter calculating, image taking etc., all operates accurately according to strict index.Therefore aforesaid method can increase substantially stability and the efficiency of order-checking process, and the accuracy of sequencing result.In addition carry out large-scale IMAQ by controlling gene sequenator, therefore ensured enough sequencing throughput.
Fig. 7 shows step S1 method flow in one embodiment in Fig. 6, the system of the method flow process based on shown in Fig. 1, Fig. 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 small chamber.If the assembly corresponding with reaction control unit 100 is the situation of describing in earlier figures 2 in gene sequencer, its control process is: reagent control module 101 is determined the required types of agents of taking of different steps, send instructions to mechanical manipulator, control mechanical manipulator and move to reagent position corresponding on reagent platform, and the flexible pipe being fixed on mechanical manipulator is inserted in reagent; Reagent control module 101 sends instructions to pump, thereby reagent is drawn in control pump running; Reagent control module 101 is drawn to after required reagent, sends instructions to pump, continues control pump running reagent is squeezed into reaction small chamber.
Step S12, the temperature of reaction small chamber is controlled at the required temperature of reaction by temperature control module 102.In the present invention there is multiple specific implementation in step S12, will be described in detail by different embodiment below.
In one embodiment, taking the situation of the gene sequencer assembly described in Fig. 2 as example, step S12 realizes by serial communication mode, detailed process is: temperature control module 102 sends instruction to temperature sensor, control temperature sensor the temperature of reaction small chamber is detected, and read temperature detection result t; In temperature control module 102, be provided with the temperature value T in differential responses stage, when it obtains after temperature detection result t, it contrasted with the temperature value T arranging; Temperature control module 102 is further processed according to comparing result, if t<T, temperature control module 102 sends instruction to the heat riser in temperature controller, the heat riser of controlling in temperature controller starts, heat to reaction small chamber, if t >=T, does not need to start heat riser heating, heat riser is cooled to temperature T automatically by outside atmosphere.
For the gene sequencer assembly under other situations, control principle is consistent, and detailed process may there are differences.For example, in another embodiment, if gene sequencer assembly is compared with the situation of describing in Fig. 2, temperature controller, except comprising for detection of the temperature sensor of reaction small chamber temperature, to the heat riser of reaction small chamber heating, also comprises the heat sink to reaction small chamber refrigeration.So in this case, the implementation procedure of step S12 is: temperature control module 102 sends instruction to temperature sensor, controls temperature sensor the temperature of reaction small chamber is detected, and read temperature detection result t; In temperature control module 102, be provided with the temperature value T in differential responses stage, when it obtains after temperature detection result t, it contrasted with the temperature value T arranging; Temperature control module 102 is further processed according to comparing result, if t<T, temperature control module 102 sends instruction to the heat riser in temperature controller, the heat riser of controlling in temperature controller starts, to reaction small chamber heating, if t>T, temperature control module 102 sends instruction to the heat sink in temperature controller, the heat sink of controlling in temperature controller starts, and reaction small chamber is freezed.
As shown in Figure 7, step S11 can be to types of agents, reagent dosage, capture velocity, get speed etc. controls accurately, step S12 can strictly control temperature detection, temperature setting, heating, refrigeration etc., thereby ensure that the biochemical reaction process in reaction small chamber carries out smoothly, also therefore improved stability, efficiency and the accuracy of whole order-checking process.
Fig. 8 shows step S2 method flow in one embodiment in Fig. 6.
Step S21, the current position of detection reaction cell in gene sequencer, and control its move to its target location in the plane.In the present invention, step S21 can control the movement of reaction small chamber in several ways.
In one embodiment, in gene sequencer, the assembly corresponding with Positioning Control Unit 200 is the situation of describing in earlier figures 2, step S21 realizes by serial communication mode: first displacement module 201 sends instruction to sample table, read sample table its starting position coordinate in the plane, be for example (X
0, Y
0); Determine reaction small chamber its after object coordinate (X, Y) in the plane, displacement module 201 sends instruction again to sample table, Quality control platform is from (X
0, Y
0) move to object coordinate (X, Y).
Adopting in figure process, for controlling gene sequenator carries out large-scale IMAQ, need to circulate and adopt figure the position one by one on DNA fragmentation sample to be measured in reaction small chamber, therefore need displacement module 201 in figure process, to control reaction small chamber and carry out displacement according to position one by one adopting.For example, starting position (X
0, Y
0), the figure position of adopting corresponding on reaction small chamber has n capable, and every row has m to adopt figure position, and each spacing of adopting between figure position equates.The capable j of i coordinate representation of adopting figure position is (X so
ij, Y
ij), the coordinate of all figure of adopting position can be expressed as: (X
11, Y
11), (X
12, Y
12) ..., (X
ij, Y
ij) ... (X
nm, Y
nm), wherein i, j, n, m are positive integer.In the time adopting figure, first displacement module 201 sends instruction to sample table, read sample table its starting position coordinate (X in the plane
0, Y
0), then sending instruction to sample table, Quality control platform is from (X
0, Y
0) move to object coordinate (X
11, Y
11); At (X
11, Y
11) position focuses on, adopts after figure completes, displacement module 201 sends instruction to sample table, and Quality control platform is from (X
11, Y
11) move to object coordinate (X
12, Y
12); At (X
12, Y
12) position focuses on, adopts after figure completes, displacement module 201 sends instruction to sample table, and Quality control platform is from (X
12, Y
12) move to object coordinate (X
13, Y
13).So move in circles, by the movement of position one by one, realize large-scale IMAQ.
Step S22, the focus adjustment of controlling gene sequenator, that determines DNA fragmentation sample to be measured in reaction small chamber adopts figure position.In the present invention, what step S22 can determine DNA fragmentation sample to be measured in several ways adopts figure position, describes below as an example of the situation of the gene sequencer assembly described in Fig. 2 example.
In one embodiment, step S22 realizes by serial communication mode: first focus module 202 sends instruction to microscope, controlling microscope is moving up with sample table Vertical Square, by regulating the distance between microscope and reaction small chamber, by the location positioning of sharpness the best for adopting figure position.
In another embodiment, step S22 remains and realizes by serial communication mode: focus module 202 sends instruction to sample table, Quality control platform moves up at the Vertical Square of its place plane, by regulating the distance between reaction small chamber and microscope, by the location positioning of sharpness the best for adopting figure position.
In above-mentioned two embodiment, step S22 can determine the position of image definition the best in several ways.For example, the mode that can observe contrast by microscope camera lens be determined the position of image definition the best, or utilizes algorithm computed image sharpness, utilizes algorithm automatically to regulate image definition, etc.
As from the foregoing, 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 determine quickly and accurately the best figure position of adopting, also therefore improve stability, efficiency and the accuracy of order-checking process.
Fig. 9 shows step S3 method flow in one embodiment in Fig. 6.
Step S31, DNA fragmentation sample to be measured in the excitation light irradiation reaction small chamber of control specific wavelength, the marker that Nucleotide is wherein carried is luminous.In the present invention, step S31 can be accomplished in several ways.
In one embodiment, in gene sequencer, the assembly corresponding with adopting figure control unit 300 is the situation of describing in earlier figures 2, the implementation procedure of step S31 is so: excitation module 301 sends instruction to light-emitting device, open shutter or optical gate, the irradiate light that makes exciting light is to the reaction small chamber in sample table.In the present embodiment, the marker that on the microballon in DNA fragmentation sample to be measured, Nucleotide carries is fluorescent marker, and it is subject to just can sending fluorescence after the light source activation of specific wavelength.
Step S32, determines the time shutter, and adopts this time shutter to take pictures to DNA fragmentation sample to be measured in reaction small chamber, obtains picture signal.In the present invention, step S32 can be accomplished in several ways.
Continue in the embodiment of abovementioned steps S31, in gene sequencer, the assembly corresponding with adopting figure control unit 300 is the situation of describing in earlier figures 2, so in step S32, first determine suitable exposure time values by photo module 302, then send instruction to CCD, control CCD and take fluorogram according to this exposure time values.
Step S32 of the present invention can determine in several ways and for example according to circumstances artificially arrange suitable exposure time values, or is set to the time shutter empirical value of repeatedly order-checking process accumulation, or calculates suitable exposure time values by algorithm, etc.In the gene sequencing Controlling System of prior art before this, major part has adopted the mode of artificial setting, and the present invention has plurality of optional pattern, is intended to determine best exposure time values according to different situations, thereby improves the quality of picture signal.
Step S33, preserves the picture signal of obtaining.In the present invention, image access module 303 can adopt multiple format memory image signal.Step S33 can be accomplished in several ways.
Continue in the embodiment of abovementioned steps S31, S32, the implementation of step S33 is: the fluorogram that photo module 302 is taken CCD sends 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, such as TIFF, EPS, PNG, PSD or binary format etc.
As from the foregoing, the light path of the exciting light that step S31 can send light-emitting device etc. is accurately controlled, step S32 can accurately control the determining of exposure time values, image taking etc., step S33 can adopt best picture format memory image signal, thereby ensure the quality of the picture signal of obtaining, greatly improved the accuracy of sequencing result, and this automatization control mode stability and the efficiency of order-checking process are also improved.
In order more clearly to explain the present invention, applicant will be as an example of a concrete experimentation example the whole process of a known gene sequencing of explanation, Figure 10 be one can be for referencial use schematic diagram.This application scenarios is to adopt a kind of known sequence measurement to check order to DNA:
(1) in connection with there being the microballon that carries the 3 ' terminal modified DNA fragmentation to be measured to be deposited on loading slide, for a large amount of microballons, be by microballon matrix point sample in slide, in point sample process, can regulate microballon density, to reach flux peak.
(2) to adding DNA ligase, universal sequencing primer thing n in reaction small chamber and thering are eight polynucleotides of 3 '-XXnnnzzz-5 ' structure.In this eight polynucleotide, the base on the 1st and the 2nd (XX) determines, and according to having added different fluorescent marks on the 6-8 position (zzz) that do not coexist of kind.This sequence measurement being determined by two bases is called as two base order-checkings (two base encoding).
(3) when eight polynucleotides are because the 1st when the 2nd bit pairing is connected enzyme and is connected, through the optical excitation of specific wavelength, can send fluorescence.
(4) recording after fluorescence information, cutting between the 5th and the 6th by chemical process, cancellation fluorescent signal, to carry out the order-checking of next position.
By this method, the position of each order-checking all differs five, surveys for the first time the 1st and the 2nd, surveys for the second time the 6th and the 7th ... measuring behind end, by newly synthetic chain sex change, wash-out.Then carry out second with universal sequencing primer thing n-1 and take turns order-checking.The difference of universal sequencing primer thing n-1 and universal sequencing primer thing n is, the two with the position of joint pairing on differ a base, universal sequencing primer thing n-1 has moved a base to 3 ' end in universal sequencing primer thing n mated position.Therefore adding after DNA ligase and eight polynucleotides, can measure the 0th and the 1st, the 5th and the 6th ... second take turns and checked order 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, fourth round, the 5th is taken turns order-checking, finally can complete the mensuration of whole positions.
Above-mentioned order-checking process, each take turns all relate to that repeatedly reagent is taken, temperature adjusting, time are controlled etc., no matter be simple manual operation, or Artificial Control instrumentation, all cannot fully ensure stability, efficiency and the accuracy of experiment.And utilize control method of the present invention and system, only the DNA fragmentation sample to be measured preparing need be arranged in the reaction small chamber of gene sequencer, select the order-checking pattern for different samples, just can make gene sequencer above-mentioned each step of operation automatically, without manual operations.After loading, through the automatic sequencing process of gene sequencer, can arrive picture signal by Quick Acquisition.It should be noted that, the whole process of the gene sequencing shown in Figure 10 is only the order-checking process under certain situation, and Controlling System and the control method of the present invention to gene sequencer do not limit which kind of order-checking flow process and sequence measurement of employing.
Should be noted that; method and system of the present invention are applicable to the order-checking process of various types of gene sequencers to carry out automatization control; even if dissimilar gene sequencer may there are differences in concrete internal structure; but above-mentioned Controlling System and control method in grammar, are consistent or similarly, therefore protection scope of the present invention should not be subject to the restriction of the internal structure of dissimilar gene sequencer.
The foregoing is only preferred embodiment of the present invention, not in order to limit the present invention, all any amendments of doing within the spirit and principles in the present invention, be equal to and replace and improvement etc., within all should being included in protection scope of the present invention.
Claims (6)
1. a system of the order-checking process of gene sequencer being controlled, wherein DNA fragmentation sample to be measured is arranged in the reaction small chamber of gene sequencer, it is characterized in that, and described system comprises reaction control unit, Positioning Control Unit, adopts figure control unit; Adopting in figure process, described system controlling gene sequenator circulates and adopts figure the position one by one on DNA fragmentation sample to be measured in reaction small chamber;
Described reaction control unit imports reaction small chamber for controlling gene sequenator by reagent, and in order-checking process, regulates the temperature of reaction small chamber;
Described Positioning Control Unit is used for controlling reaction small chamber and successively moves at gene sequencer, and determine each mobile after in reaction small chamber DNA fragmentation sample to be measured adopt figure position;
The described figure of adopting control unit is luminous for the marker that excites DNA fragmentation sample Nucleotide to be measured to carry, and after described each movement in reaction small chamber DNA fragmentation sample to be measured adopt figure position acquisition picture signal;
Described reaction control unit comprises reagent control module, temperature control module;
Described reagent control module is selected reagent for controlling gene sequenator, and draws corresponding reagent, imports reaction small chamber;
Described temperature control module is for being controlled at the temperature of reaction small chamber the required temperature of reaction;
The concrete control process of described temperature control module is: temperature control module sends instruction to temperature sensor, controls temperature sensor the temperature of reaction small chamber is detected, and read temperature detection result t; In temperature control module, be provided with the temperature value T in differential responses stage, when it obtains after temperature detection result t, it contrasted with the temperature value T arranging; Temperature control module is further processed according to comparing result, if t<T, temperature control module sends instruction to the heat riser in temperature controller, the heat riser of controlling in temperature controller starts, to reaction small chamber heating, if t>T, temperature control module sends instruction to the heat sink in temperature controller, the heat sink of controlling in temperature controller starts, and reaction small chamber is freezed;
The described figure of adopting control unit comprises excitation module, photo module, image access module;
Described excitation module, for controlling the excitation light irradiation reaction small chamber DNA fragmentation sample to be measured of specific wavelength, makes the marker that in DNA fragmentation sample to be measured, Nucleotide carries luminous;
Described photo module calculates suitable exposure time values by time shutter empirical value or the algorithm of the accumulation of repeatedly order-checking process, then send instruction to CCD, control CCD and according to this exposure time values, DNA fragmentation sample to be measured in reaction small chamber is taken pictures, obtain fluorescence image signal;
Described image access module and photo module communicate, for preserving the picture signal of obtaining.
2. the system that the order-checking process of gene sequencer is controlled according to claim 1, is characterized in that, described Positioning Control Unit comprises displacement module, focus module;
Described displacement module is the current position in gene sequencer for detection of reaction small chamber, and control its move to its target location in the plane;
Described focus module is for the focus adjustment of controlling gene sequenator, and that determines DNA fragmentation sample to be measured in reaction small chamber adopts figure position.
3. the system that the order-checking process of gene sequencer is controlled according to claim 2, is characterized in that, described focus module is by regulating the distance between microscope and reaction small chamber, by the location positioning of sharpness the best for adopting figure position.
4. a method of the order-checking process of gene sequencer being controlled based on system described in claim 1, wherein DNA fragmentation sample to be measured is arranged in the reaction small chamber of gene sequencer, it is characterized in that, adopting in figure process, circulated and adopt figure in position one by one on DNA fragmentation sample to be measured in reaction small chamber, said method comprising the steps of:
A. reagent is imported reaction small chamber by controlling gene sequenator, and regulate the temperature of reaction small chamber;
B. control reaction small chamber and successively move in gene sequencer, and determine each mobile after in reaction small chamber DNA fragmentation sample to be measured adopt figure position;
C. excite the marker that in DNA fragmentation sample to be measured, Nucleotide carries luminous, and after described each movement in reaction small chamber DNA fragmentation sample to be measured adopt figure position acquisition picture signal;
Described steps A comprises:
A1. controlling gene sequenator is selected reagent, and draws corresponding reagent, imports reaction small chamber;
A2. the temperature of reaction small chamber is controlled to the required temperature of reaction;
The implementation procedure of described steps A 2 is: temperature control module sends instruction to temperature sensor, controls temperature sensor the temperature of reaction small chamber is detected, and read temperature detection result t; In temperature control module, be provided with the temperature value T in differential responses stage, when it obtains after temperature detection result t, it contrasted with the temperature value T arranging; Temperature control module is further processed according to comparing result, if t<T, temperature control module sends instruction to the heat riser in temperature controller, the heat riser of controlling in temperature controller starts, to reaction small chamber heating, if t>T, temperature control module sends instruction to the heat sink in temperature controller, the heat sink of controlling in temperature controller starts, and reaction small chamber is freezed;
Described step C comprises:
C1. control DNA fragmentation sample to be measured in the excitation light irradiation reaction small chamber of specific wavelength, make the marker that in DNA fragmentation sample to be measured, Nucleotide carries luminous;
C2. calculate suitable exposure time values by time shutter empirical value or the algorithm of the accumulation of repeatedly order-checking process, then send instruction to CCD, control CCD and according to this exposure time values, DNA fragmentation sample to be measured in reaction small chamber is taken pictures, obtain fluorescence image signal;
C3. preserve the picture signal of obtaining.
5. the method that the order-checking process of gene sequencer is controlled according to claim 4, is characterized in that, described step B comprises:
B1. the current position of detection reaction cell in gene sequencer, and control its move to its target location in the plane;
B2. the focus adjustment of controlling gene sequenator, that determines DNA fragmentation sample to be measured in reaction small chamber adopts figure position.
6. the method that the order-checking process of gene sequencer is controlled according to claim 5, is characterized in that, described step B2 comprises:
By regulating the distance between microscope and reaction small chamber, by the location positioning of sharpness the best for adopting figure position.
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| CN102517206B (en) * | 2011-12-31 | 2015-06-03 | 盛司潼 | Gene sequencing device and system |
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| CN106434272B (en) * | 2015-08-07 | 2019-06-21 | 广州康昕瑞基因健康科技有限公司 | Gene sequencer, self-level(l)ing device and method |
| CN105319156B (en) * | 2015-12-02 | 2017-07-11 | 北京中科紫鑫科技有限责任公司 | It is a kind of that sequencing system is gathered based on the DNA image that CCD camera is adjusted |
| CN106250720B (en) * | 2016-08-23 | 2019-08-09 | 山东卫康医学检验有限公司 | A kind of DNA sequencing image processing system being grouped in advance according to similarity |
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| CN107871059A (en) * | 2016-09-23 | 2018-04-03 | 广州康昕瑞基因健康科技有限公司 | Gene sequencer runs analogy method and system |
| CN108085378A (en) * | 2016-11-16 | 2018-05-29 | 广州康昕瑞基因健康科技有限公司 | Gene sequencer progress display adjusting method and system |
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