CN207215686U - Systems for optical inspection and Sequence Detection System - Google Patents

Abstract

The utility model discloses a kind of Systems for optical inspection and Sequence Detection System.Systems for optical inspection includes control device, imaging device and bogey, bogey includes temperature control device and microscope carrier, imaging device includes camera lens module, camera lens module includes optical axis, microscope carrier carrying sample, and Systems for optical inspection is preset with the moving range of camera lens module, moving range is determined by the first setting position and the second setting position, the setting of second setting position is relevant with the position of sample and the depth of field, and temperature control device adjusts the temperature of sample, and control device is used for：Before IMAQ is carried out to sample using imaging device or when carrying out IMAQ to sample using imaging device, the scope for the temperature fluctuation for allowing sample is set using temperature control device, so that positional fluctuation scope of the camera lens module along optical axis is located in preset range.In this way, using above-mentioned Systems for optical inspection, adverse effect when IMAQ is carried out to imaging device can be reduced or avoided.

Description

Systems for optical inspection and Sequence Detection System

Technical field

It the utility model is related to field of optical detection, more particularly to a kind of Systems for optical inspection and a kind of sequencing system System.

Background technology

Sequencing, that is, it is sequenced, includes the measure of nucleotide sequence.It is flat to include generation sequencing for microarray dataset on the market at present Platform, two generation microarray datasets and three generations's microarray dataset.From function control angle, sequencing instrument includes detecting module, using detecting mould Block converts and/or collected information change caused by the reaction of sequencing mesophytization, to determine sequence.Detecting module generally comprises light Learn detection module, current detection module and soda acid (pH) detection module.Microarray dataset based on optical detection principle passes through analysis Acquisition testing to sequencing biochemical reaction in change in optical signal carry out sequencing.

During optical signal is gathered, the camera lens module of optical detecting module can be expected outside positional fluctuation, this Collection of the kind positional fluctuation to optical signal brings adverse effect.

Utility model content

The utility model embodiment is intended at least solve one of technical problem present in correlation technique or at least carried For a kind of selectable practical plan.Therefore, the utility model embodiment needs to provide a kind of Systems for optical inspection and one kind Sequence Detection System.

Inventor is based on the finding that and produce the design for making the utility model：

Those skilled in the art understand that, during imaging, image/signal acquisition or sequencing, particularly wrap Contained the platform of the optical system of high accuracy/high magnification numbe, the camera lens module (including object lens) of Systems for optical inspection if occur shake/ Fluctuation or the shake/fluctuation occurred are uncontrolled, it is likely that influence the collection of image, influence the acquisition of signal.

In microarray dataset using imaging device against reaction unit (such as chip, flow cell) carry out chasing after Jiao when, hair A person of good sense it has surprisingly been found that the coordinate (being set to the coordinate in Z-direction) of camera lens module (including object lens) along chasing after burnt direction (such as core The channel direction of piece, it is determined as x directions) change curve (z-x) do not meet theoretic linear relationship or be similar to linear Relation, violent localised waving is showed, and periodically change is presented in the fluctuation, as shown in Figure 1.Further, invent People tests, even if without chasing after the burnt i.e. not instruction to the motion of camera lens module Z-direction, only by camera lens module against the same visual field Taken pictures, camera lens module Z-direction still cyclic fluctuation, as shown in Fig. 2 the cycle is about 10.4s.More in surprise, invent People has found that the cycle of the fluctuation period of change of the temperature with allowing reaction unit is very close, and the fluctuation of guess camera lens module can Can be relevant with the fluctuation of temperature.Usually, it is allowed to which the setting of the temperature change of reaction unit with reaction unit by being connected What temperature control device was controlled, such as wish that the temperature of reaction unit is maintained at 25 DEG C or so, temperature can be set by temperature control device It is [24 DEG C, 26 DEG C] to spend excursion, i.e., beyond 25 ± 1 DEG C, temperature control device maintains the temperature of reaction unit by temperature is adjusted In above-mentioned preset range.

Inventor removes by controlling the above-mentioned guess of temperature fluctuation substantive test, such as by temperature control device, along chip One passage carries out being similar to the behavior for chasing after Jiao back and forth, and camera lens module Z-X fluctuations meet theoretic linear or approximately linear Relation, do not occur cyclic fluctuation phenomenon described above.Discovery and confirmation based on relation above, inventor propose a kind of profit The scheme that camera lens module fluctuates is controlled with control temperature change.

A kind of Systems for optical inspection of the utility model embodiment includes control device, imaging device and bogey, The bogey includes temperature control device and microscope carrier, and the imaging device includes camera lens module, and the camera lens module includes optical axis, The microscope carrier is used to carry sample, and the Systems for optical inspection is preset with the moving range of the camera lens module, the mobile model Enclose and determined by the first setting position and the second setting position, the setting of second setting position and the position of the sample with And the depth of field is relevant, the temperature control device is used for the temperature for adjusting the sample, and the control device is used for：Utilizing the imaging Before device carries out IMAQ to the sample or when carrying out IMAQ to the sample using the imaging device, Allow the scope of the temperature fluctuation of the sample using temperature control device setting, so that the camera lens module is along the optical axis Positional fluctuation scope is located in preset range.

A kind of Sequence Detection System of the utility model embodiment, sequencing reaction is controlled, the sequence Measurement system includes Systems for optical inspection, and the Systems for optical inspection includes control device, imaging device and bogey, described Imaging device includes camera lens module, and the camera lens module includes optical axis, and the bogey includes temperature control device and microscope carrier, described Microscope carrier is used to carry sample, and the Systems for optical inspection is preset with the moving range of the camera lens module, the moving range by Determine that second setting position is set and the position of the sample and scape with the second setting position in the first setting position Deep relevant, the temperature control device is used for the temperature for adjusting the sample, and the control device is used to utilize the imaging device pair The sample carries out IMAQ, and is used for：

Before the sequencing reaction is carried out using the Sequence Detection System or utilizing the sequencing When system carries out the sequencing, the scope of the temperature fluctuation of the sample is allowed using temperature control device setting, so that Positional fluctuation scope of the camera lens module along the optical axis is located in preset range.

, can be by the positional fluctuation scope control of camera lens module using above-mentioned Systems for optical inspection and/or Sequence Detection System Within a preset range, adverse effect when IMAQ is carried out to imaging device is reduced or avoided in system.

The additional aspect and advantage of the utility model embodiment will be set forth in part in the description, partly will be under Become obvious in the description in face, or recognized by the practice of the utility model embodiment.

Brief description of the drawings

The above-mentioned and/or additional aspect and advantage of the utility model embodiment are from accompanying drawings below is combined to embodiment Description in will be apparent and be readily appreciated that, wherein：

Fig. 1 is that the Z axis coordinate edge of the camera lens module detected chases after the change curve schematic diagram in burnt direction.

Fig. 2 is that the Z axis coordinate edge of the camera lens module detected chases after another change curve schematic diagram in burnt direction.

Fig. 3 is the schematic perspective view of the bogey of the utility model embodiment.

Fig. 4 is the decomposing schematic representation of the bogey of the utility model embodiment.

Fig. 5 is the camera lens module of the utility model embodiment and the position relationship schematic diagram of sample.

Fig. 6 is the part-structure schematic diagram of the Systems for optical inspection of the utility model embodiment.

Fig. 7 is the schematic flow sheet of the imaging method of the utility model embodiment.

Fig. 8 is another schematic flow sheet of the imaging method of the utility model embodiment.

Fig. 9 is another schematic flow sheet of the imaging method of the utility model embodiment.

Figure 10 is the decomposing schematic representation of the temperature control device of the utility model embodiment.

Figure 11 is water-bath room and the structural representation of temperature controlled water bath device of the utility model embodiment.

Figure 12 is the decomposed schematic diagram of the bogey of the utility model embodiment.

Figure 13 is the structural representation of the fluid means of the utility model embodiment.

Figure 14 is the schematic flow sheet of the method for the control sequence measure reaction of the utility model embodiment.

Figure 15 is the module diagram of the imaging device of the utility model embodiment.

Figure 16 is another module diagram of the imaging device of the utility model embodiment.

Figure 17 is the module diagram of the Systems for optical inspection of the utility model embodiment.

Figure 18 is the module diagram of the Sequence Detection System of the utility model embodiment.

Embodiment

Embodiment of the present utility model is described below in detail, the example of the embodiment is shown in the drawings, wherein Same or similar label represents same or similar element or the element with same or like function from beginning to end.Lead to below It is exemplary to cross the embodiment being described with reference to the drawings, and is only used for explaining the utility model, and it is not intended that to this practicality New limitation.

In description of the present utility model, it is to be understood that term " first ", " second " are only used for describing purpose, and It is not intended that instruction or hint relative importance or the implicit quantity for indicating indicated technical characteristic.Thus, define One or more feature can be expressed or be implicitly included to the feature of " first ", " second ".In the utility model Description in, " multiple " are meant that two or more, unless otherwise specifically defined.

, it is necessary to which explanation, unless otherwise clearly defined and limited, " connection " should be done in description of the present utility model It broadly understood, for example, it may be being fixedly connected or being detachably connected, or be integrally connected；Can mechanically connect, Can also electrically connect or can be in communication with each other；Can be joined directly together, can also be indirectly connected by intermediary, Ke Yishi The connection of two element internals or the interaction relationship of two elements.For the ordinary skill in the art, can be with Concrete meaning of the above-mentioned term in the utility model is understood as the case may be.

In description of the present utility model, it is to be understood that term " thickness ", " on ", " under ", "front", "rear", " bottom " The orientation or position relationship of the instructions such as " interior ", " outer " are based on orientation shown in the drawings or position relationship, are for only for ease of and retouch State the utility model and simplify and describe, rather than instruction or imply signified device or element must have specific orientation, with Specific azimuth configuration and operation, therefore it is not intended that to limitation of the present utility model.

In description of the present utility model, alleged " constant ", such as it is related to distance, object distance and/or relative position etc. The change in numerical value, number range or amount can be shown as, can be definitely constant or relatively constant, alleged phase To not being changed into being maintained at certain deviation range or default tolerance interval.Unless otherwise noted, be related to distance, object distance and/ Or relative position is " constant " to be relatively constant.It should be noted that due to the specific number being related in description of the present utility model According to having statistical significance mostly, therefore, unless otherwise specified, the numerical value arbitrarily expressed in a precise way represents a scope, The section of the numerical value positive and negative 10% is included, is not repeated to illustrate below.

" sequencing " same determining nucleic acid sequence alleged by the utility model embodiment, including DNA sequencing and/or RNA Sequencing, including long segment sequencing and/or short-movie section sequencing.Alleged " sequencing reaction " same to sequencing reaction.

The utility model embodiment provides a kind of imaging method, and imaging method is used for Systems for optical inspection, optical detection System includes imaging device 102 and bogey 100, and bogey 100 includes temperature control device 301 and microscope carrier 103, imaging device 102 include camera lens module 104, and camera lens module 104 includes optical axis OP, and microscope carrier 103 is used to carry sample 300, and temperature control device 301 is used In the temperature of regulation sample 300.Imaging method includes：Using imaging device 102 to sample 300 carry out IMAQ before or Person sets the temperature for allowing sample 300 using temperature control device 301 when carrying out IMAQ to sample 300 using imaging device 102 The scope of fluctuation is spent, so that positional fluctuation scope of the camera lens module 104 along optical axis OP is located in preset range.Utilize above-mentioned imaging Method, it can be reduced or avoided by the positional fluctuation scope control of camera lens module 104 within a preset range to imaging device 102 Carry out adverse effect during IMAQ.

Specifically, temperature control device 301 can be contacted with sample 300 to set the scope of the temperature fluctuation of sample 300.

In some embodiments, Systems for optical inspection is preset with the scope and camera lens module of the temperature fluctuation of sample 300 104 along the positional fluctuation scope of optical axis corresponding relation, according to corresponding relation, control camera lens module 104 is along optical axis OP position Fluctuation range is located in preset range.In this way, imaging method can be obtained positional fluctuation of the camera lens module 104 along optical axis rapidly The scope of the scope control temperature fluctuation of corresponding sample 300 within a preset range.

Specifically, the scope of the temperature fluctuation of sample 300 is corresponding with positional fluctuation scope of the camera lens module 104 along optical axis Relation is storable in Systems for optical inspection.In one example, alleged corresponding relation can join table 1 below.

Table 1

It is pointed out that the data of above-mentioned table 1 are intended only as example to illustrate the utility model embodiment, it is impossible to manage Solve as to limitation of the present utility model.In other examples, the scope of the temperature fluctuation to sample 300 can be carried out according to the actual requirements Specific setting corresponding relation is carried out with positional fluctuation scope of the camera lens module 104 along optical axis.

In some embodiments, the structure of bogey 100 is as shown in Figure 3 and Figure 4.Camera lens module 104 and sample 300 Position relationship it is as shown in Figure 5.

Again in some embodiments, Fig. 6 please be join, imaging device 102 includes focusing module 106, right using imaging device 102 Sample 300 carries out IMAQ, including：Using focusing module 106 and camera lens module 104 is focused to sample 300.In this way, Imaging device 102 is focused in IMAQ, can get clearly sample image.

In some embodiments, Fig. 7 please be join, focusing includes step：S11, emitted light into and be placed in using module 106 of focusing On sample 300 on microscope carrier 103；S12, camera lens module 104 is set to be moved to the first setting position along optical axis OP；S13, make camera lens mould Group 104 is moved along optical axis 300 to sample 300 with the first setting step-length from the first setting position and whether judges focusing module 106 Receive the light reflected by sample 300；When module 106 of focusing receives the light reflected by sample 300, S14, make camera lens module 104 are moved along optical axis OP with the second setting step-length less than the first setting step-length and sample 300 are carried out using imaging device 102 IMAQ, and judge whether the sharpness value for the image that imaging device 102 is collected reaches given threshold；In the acutance of image When value reaches given threshold, S15, the current location of camera lens module 104 is preserved as save location.Using above-mentioned imaging method, The plane of target object blur-free imaging, i.e., clear plane/clear face can rapidly and accurately be found.This method is not especially suitable for Easily find the equipment for including precision optical system of clear plane, such as the optical detection apparatus with high magnification numbe camera lens.

Specifically, sample 300 can be regarded as sensu lato sample or sample in the narrow sense.For sensu lato sample, sample Product are the testing samples such as every solution, reagent.Sample can be placed on supporting arrangement, supporting arrangement is, for example, that reaction unit is (such as Chip), slide etc., the supporting arrangement with sample is placed on bogey 100.For sample in the narrow sense, sample includes Supporting arrangement and the testing sample on supporting arrangement, during sequencing, sample can be placed on bogey 100.Please Join Fig. 5, in the embodiment of focusing step, sample 300 includes supporting arrangement 200 and treats test sample positioned at supporting arrangement 200 Product 302, testing sample 302 are biomolecule, such as nucleic acid, and camera lens module 104 is located at the top of supporting arrangement 200.Branch takes up Putting 200 has front panel 202 and rear board (lower panel), and each panel is respectively provided with two surfaces, and testing sample 302 is connected to following On the upper surface of plate, i.e., testing sample 302 is located at the lower section of lower surface 204 of front panel 202.In the utility model embodiment In, because imaging device 102 is the image of collection testing sample 302, and testing sample 302 is located at before supporting arrangement 200 The lower section of 202 lower surface of plate 204, when focus process starts, the movement of camera lens module 104 is to find the institute of testing sample 302 Dielectric interface 204, with improve imaging device 102 collection picture rich in detail success rate.In the utility model embodiment party In formula, testing sample 302 is solution, and the front panel 202 of supporting arrangement 200 is glass, supporting arrangement 200 and testing sample 302 Dielectric interface 204 for the lower surface 204, i.e. glass and liquid two media of the front panel 202 of supporting arrangement 200 boundary Face.The testing sample 302 of collection image is located under the lower surface 204 of front panel 202 needed for imaging device 102, now again The clear face of the searching blur-free imaging of testing sample 302 is differentiated by the acquired image of imaging device 102, this process can be described as Focusing.In one example, the thickness of the front panel 202 of testing sample 302 is 0.175mm.

In some embodiments, supporting arrangement 200 can be slide, and testing sample 302 is placed on slide, or treats test sample Product 302 are located in two panels slide.In some embodiments, supporting arrangement 200 can be reaction unit, for example, holding up and down The chip similar to sandwich structure of section plate, testing sample 302 are arranged on chip.

In some embodiments, Fig. 6 please be join, imaging device 102 includes microscope 107 and camera 108, camera lens module 104 include the camera lens module 112 of microscopical object lens 110 and camera 108, and focusing module 106 can pass through dichroic beam splitters 114 (dichroic beam splitter) and camera 108 camera lens module 112 are fixed together, and dichroic beam splitters 114 are located at Between the camera lens module 112 and object lens 110 of camera 108.Dichroic beam splitters 114 include double C-type beam splitter (dual c-mount splitter).Dichroic beam splitters 114 can reflect the light of the focusing transmitting of module 106 to object lens 110 and visible ray can be allowed to penetrate And the camera lens module 112 through camera 108 enters in camera 108, as shown in Figure 6.

In the utility model embodiment, the movement of camera lens module 104 can refer to the movement of object lens 110, camera lens module 104 Position can refer to the positions of object lens 110.In other embodiments, other lens that moving lens module 104 may be selected come in fact Now focus.In addition, microscope 107 also includes the tube lens 111 (tube lens) between object lens 110 and camera 108.

In some embodiments, microscope carrier 103 can drive sample 200 in the optical axis OP perpendicular to camera lens module 104 (such as Z axis) plane in move (such as X/Y plane), and/or optical axis OP (such as Z axis) of the sample 300 along camera lens module 104 can be driven to move It is dynamic.

In some embodiments, microscope carrier 103 drives the plane that sample 300 moves non-perpendicular to optical axis OP, i.e. sample Plane of movement and X/Y plane angle are non-zero, and the imaging method is still applicable.

In addition, imaging device 102 can also drive optical axis OP of the object lens 110 along camera lens module 104 to move to be focused. In some instances, imaging device 102 drives object lens 110 to move using actuators such as stepper motor or voice coil motors.

In some embodiments, when establishing coordinate system, as shown in figure 5, can be by object lens 110, microscope carrier 103 and sample 300 position is arranged on the negative axle of Z axis, and the first setting position can be the coordinate position on the negative axle of Z axis.It is appreciated that In other embodiment, the relation of coordinate system and camera and object lens 110 can be also adjusted according to actual conditions, do not done herein It is specific to limit.

In one example, imaging device 102 includes utilizing total internal reflection fluorescence microscope, and object lens 110 are 60 times and amplified, and first Set step-length S1=0.01mm.In this way, the first setting step-length S1 is appropriate, because S1 too conferences are across acceptable focusing range, S1 is too small to increase time overhead.

When module 106 of focusing does not receive the light reflected by sample 300, then camera lens module 104 is set to be walked with the first setting Length continues to move to along optical axis OP to sample 300.

In some embodiments, when the sharpness value of image does not reach given threshold, then camera lens module 104 is made with second Setting step-length continues to move to along optical axis OP.

In some embodiments, Systems for optical inspection can be applied to Sequence Detection System, in other words, Sequence Detection System Including Systems for optical inspection.

In some embodiments, when camera lens module 104 moves, judge whether the current location of camera lens module 104 surpasses Go out the second setting position；When exceeding the second setting position in the current location of camera lens module 104, stop moving lens module 104 Or carry out focusing step.In this way, the first setting position can limit the moving range of camera lens module 104 with the second setting position, Camera lens module 104 can be made to stop movement when that can not focus successfully, avoid the waste of resource or the damage of equipment, or can make Camera lens module 104 is focused again when that can not focus successfully, improves the automation of imaging method.Camera lens module 104 Moving range can be preset and be preserved by Systems for optical inspection.

In some embodiments, such as in total internal reflection imaging system, for dielectric interface can be quickly found out, can adjust Whole set makes the moving range of camera lens module 104 as far as possible small in the case where that can meet to implement the program.For example, it is 60 in object lens On total internal reflection imaging device again, according to light path characteristic and summary of experience, the moving range of camera lens module 104 may be configured as 200 μm ± 10 μm or be [190 μm, 250 μm].

In some embodiments, the moving range of camera lens module 104 is true by the first setting position and the second setting position Make, the setting of the second setting position is relevant with the position of sample 300 and the depth of field.According to fixed moving range and second The setting of any position in setting position and the first setting position, it may be determined that another setting position.In one example, second set It is depth of field when be imaged according to imaging device 102 to sample 300 to set that positioning, which is put, specifically, in the present embodiment, is set Second setting position for the front panel 202 of reaction unit 200 the lowest part of upper surface 205 again toward the position of next depth of field size, The moving range of camera lens module 104 is set as 250 μm, in this way, the first setting position determines.In the utility model example, Coordinate position corresponding to the position of next depth of field size is the position to diminish along Z axis negative direction.

Specifically, in the utility model embodiment, moving range is a section on the negative axle of Z axis.At one In example, the first setting position is nearlimit, and the second setting position is farlimit, nearlimit and farlimit corresponding Coordinate position be respectively positioned on the negative axle of Z axis, nearlimit=-6000um, farlimit=-6350um.Nearlimit and The size of the moving range limited between farlimit is 350um.Therefore, when seat corresponding to the current location of camera lens module 104 Cursor position is less than the current location that camera lens module 104 during coordinate position, is judged corresponding to the second setting position beyond the second setting Position.In Figure 5, farlimit position is the next depth of field L of the lowest part of upper surface 205 of the front panel 202 of reaction unit 200 Position.Depth of field L is the depth of field size of camera lens module 104.

It is pointed out that in other embodiments, the seat corresponding to the first setting position and/or the second setting position Cursor position can make specific setting according to actual conditions, be not especially limited herein.

In some embodiments, focusing module 106 includes the first light source 116 and optical sensor 118, the first light source 116 For emitting light into sample 300, optical sensor 118 is used to receive the light reflected by sample 300.In this way, focusing mould can be realized The luminous and reception light of group 106.

Specifically, in the utility model embodiment, the first light source 116 can be infrared light supply 116, optical sensor 118 Can be photodiode (photodiode), in this way, cost is low, the accuracy rate of detection is high.The infrared light of first light source 116 transmitting Object lens 110 are reflected into through dichroic beam splitters, and sample 300 is projected through object lens 110.Sample 300 can be reflected through object lens The infrared light of 110 projections.In the embodiment of focusing step, when sample 300 includes supporting arrangement 200 and testing sample 302 When, the light just reflected by the lower surface 204 of the front panel of supporting arrangement 200 of the reflection of sample 300 of reception.

Can the infrared light that sample 300 reflects enter object lens 110 and be received by optical sensor 118, depend primarily on thing The distance of mirror 110 and sample 300.Therefore, when judging that focusing module 106 receives the infrared light of the reflection of sample 300, can determine whether The distance of object lens 110 and sample 300 is in optical imagery OK range, can be used in the imaging of imaging device 102.At one In example, distance is 20-40um.

Now, camera lens module 104 is made with the second setting step-length movement less than the first setting step-length so that optical detection system System can find the optimal imaging position of camera lens module 104 in smaller scope.

In some embodiments, the evaluation of estimate (evaluation that the sharpness value of image can focus as image value).In one embodiment, judge whether the sharpness value for the image that imaging device 102 gathers reaches given threshold and can lead to Cross the hill-climbing algorithm of image procossing.By the acutance for calculating the image that imaging device 102 is exported at each position of object lens 110 Value judges whether sharpness value reaches the maximum at sharpness value crest, and then judges whether camera lens module 104 reaches imaging dress The position where clear face when putting 102 imaging.It is appreciated that in other embodiments, other image procossings can also be used Algorithm judge whether sharpness value reaches the maximum at crest.

When the sharpness value of image reaches given threshold, the current location of camera lens module 104 is preserved as save location, can So that when sequencing reacts progress IMAQ, imaging device 102 can export clearly image.

In some embodiments, Fig. 8 please be join, when module 106 of focusing receives the light reflected by sample 300, focusing Also include step：S16, camera lens module 104 is set to be walked with the 3rd setting less than the first setting step-length and more than the second setting step-length Length moves along optical axis OP to sample 300, and the light intensity of the light received according to focusing module 106 calculates the first light intensity parameter, Judge whether the first light intensity parameter is more than the first setting Intensity threshold；It is more than the first setting Intensity threshold in the first light intensity parameter When, carry out step S14.In this way, by the comparison of the first light intensity parameter and the first setting Intensity threshold, can exclude to demarcate with medium Face reflected light contrasts very weak optical signal to interference caused by focusing/focusing.

When the first light intensity parameter is no more than the first setting Intensity threshold, then make camera lens module 104 with the 3rd setting step-length Continued to move to along optical axis OP to sample 300.

In some embodiments, focusing module 106 includes two optical sensors 118, and two optical sensors 118 are used to connect The light reflected by sample 300 is received, the first light intensity parameter is the average value of the light intensity for the light that two optical sensors 118 receive.Such as This, the average value of the light intensity of the light received by two optical sensors 118 calculates the first light intensity parameter so that excludes weak Optical signal is more accurate.

Specifically, the first light intensity parameter may be configured as SUM, i.e. SUM=(PD1+PD2)/2, PD1 and PD2 represent two respectively The light intensity for the light that individual optical sensor 118 receives.In one example, the first setting Intensity threshold nSum=40.

In one example, the 3rd setting step-length S2=0.005mm.It is appreciated that in other examples, the 3rd setting step Length can also use other numerical value, be not especially limited herein.

In some embodiments, Fig. 9 please be join, when module 106 of focusing receives the light reflected by sample 300, focusing It is further comprising the steps of：S16, camera lens module 104 is set to be set with the 3rd less than the first setting step-length and more than the second setting step-length Fixed step size moves along optical axis OP to sample 300, and the light intensity of the light received according to focusing module 106 calculates the first light intensity ginseng Number, judges whether the first light intensity parameter is more than the first setting Intensity threshold；It is more than the first setting light intensity threshold in the first light intensity parameter During value, S17, camera lens module 104 is set to set step-length along light with the 4th less than the 3rd setting step-length and more than the second setting step-length Axle OP moves to sample 300, and the light intensity of light received according to focusing module 106 calculates the second light intensity parameter, judges the Whether two light intensity parameters are less than the second setting Intensity threshold；When the second light intensity parameter is less than the second setting Intensity threshold, carry out Step S14.In this way, by the comparison of the first light intensity parameter and the first setting Intensity threshold, can exclude to reflect with dielectric interface Light contrasts very weak optical signal to interference caused by focusing/focusing；And pass through the second light intensity parameter and the second setting light intensity threshold The comparison of value, the strong reflection signal of non-dielectric interface position, such as the light letter of 110 pastas of object lens/air reflection can be excluded Number to caused by focusing/focusing interference.

When the first light intensity parameter is no more than the first setting Intensity threshold, then make camera lens module 104 with the 3rd setting step-length Continued to move to along optical axis OP to sample 300.When the second light intensity parameter is not less than the second setting Intensity threshold, then make camera lens module 104 are continued to move to the 4th setting step-length along optical axis OP to sample 300.

In one example, the 3rd setting step-length S2=0.005mm, the 4th setting step-length S3=0.002mm.It can manage Solution, in other examples, the 3rd setting step-length and the 4th setting step-length can also use other numerical value, be not especially limited herein.

In some embodiments, focusing module 106 includes two optical sensors 118, and two optical sensors 118 are used to connect The light that receipts are reflected by sample 300, the average value of the light intensity for the light that the first light intensity parameter receives for two optical sensors 118, two The light intensity for the light that individual optical sensor 118 receives has the first difference, and the second light intensity parameter is the first difference and setting compensation value Difference.In this way, the light intensity of the light received by two optical sensors 118 calculates the second light intensity parameter so that excludes strong The optical signal of reflection is more accurate.

Specifically, the first light intensity parameter may be configured as SUM, i.e. SUM=(PD1+PD2)/2, PD1 and PD2 represent two respectively The light intensity for the light that individual optical sensor 118 receives.In one example, the first setting Intensity threshold nSum=40.Difference can be set Err is set to, setting compensation value is offset, i.e. err=(PD1-PD2)-offset.In the ideal situation, the first difference can be Zero.In one example, the second setting Intensity threshold nErr=10, offset=30.

In some embodiments, when making camera lens module 104 with the second setting step-length movement, camera lens module 104 is judged Current location corresponding to the first sharpness value of pattern whether be more than image corresponding to the prior location of camera lens module 104 Second sharpness value；It is big more than the acutance difference between the second sharpness value and the first sharpness value and the second sharpness value in the first sharpness value When difference is set, camera lens module 104 is set to continue to move to sample 300 along optical axis OP with the second setting step-length；In the first acutance When the acutance difference that value is more than between the second sharpness value and the first sharpness value and the second sharpness value is less than setting difference, make camera lens mould Group 104 continues to move so that imaging device 102 along optical axis OP to sample 300 with the 5th setting step-length less than the second setting step-length The sharpness value of the image collected reaches given threshold；It is more than the first sharpness value and the second sharpness value and the in the second sharpness value When acutance difference between one sharpness value is more than setting difference, make camera lens module 104 remote along optical axis OP with the second setting step-length Sample 300 moves；It is more than the acutance difference between the first sharpness value and the second sharpness value and the first sharpness value in the second sharpness value During less than setting difference, camera lens module 104 is set to be moved with the 5th setting step-length along optical axis OP away from sample 300 so that imaging device The sharpness value of 102 images collected reaches given threshold.It so, it is possible accurately to find at sharpness value crest and correspond to Camera lens module 104 position, the image clearly for being exported imaging device.

Specifically, the second setting step-length can be used as coarse adjustment steps Z1, and the 5th setting step-length can be used as fine tuning step-length Z2, and can Coarse adjustment range Z3 is set.When coarse adjustment range Z3 setting can make the sharpness value of image can not reach given threshold, mirror can be stopped The movement of head mould group 104, has saved resource.

Using the current location of camera lens module 104 as starting point T, coarse adjustment range Z3 is adjusting range, i.e., the adjustment on Z axis Scope is (T, T+Z3).First made with step-length Z1 in the range of (T, T+Z3) camera lens module 104 in the first direction (such as along optical axis OP to The close direction of sample 300) it is mobile, and compare the figure that imaging device 102 is collected at the current location of camera lens module 104 First sharpness value R1 of picture and the second acutance of the image that imaging device 102 is collected in prior location of camera lens module 104 Value R2.

Work as R1>R2 and R1-R2>During R0, that is, illustrate that the sharpness value of image is close to given threshold and from given threshold farther out, Camera lens module 104 is set to continue to move along the first direction with step-length Z1, with rapidly close to given threshold.

Work as R1>R2 and R1-R2<During R0, that is, illustrate that the sharpness value of image is close to given threshold and nearer from given threshold, Camera lens module 104 is set to be moved along the first direction with step-length Z2, it is close to given threshold with less step-length.

Work as R2>R1 and R2-R1>During R0, that is, illustrate the sharpness value of image across given threshold and from given threshold farther out, Make camera lens module 104 with the step-length Z1 edges second direction opposite with first direction (such as along directions of the optical axis OP away from sample 300) It is mobile, with rapidly close to given threshold.

Work as R2>R1 and R2-R1<During R0, that is, illustrate that the sharpness value of image is across given threshold and nearer from given threshold, Camera lens module 104 is moved with step-length Z2 along the second direction opposite with first direction, leaned on less step-length to given threshold Closely.

In some embodiments, when camera lens module 104 moves, the 5th setting step-length can be adjusted to adapt to setting Determine threshold value close to when step-length should not be too big or too small.

In one example, T=0, Z1=100, Z2=40, Z3=2100, adjusting range are (0,2100).Need to illustrate , above-mentioned numerical value is used in imaging device 102 and carries out in image acquisition process measurement used during to moving lens module 104 Value, the metric are that light intensity is related.

In some embodiments, IMAQ is carried out to sample 300 using imaging device 102, including：Utilize camera lens mould 104 pairs of samples 300 of group carry out chasing after Jiao.So, it is ensured that imaging device 102 is adopted when the diverse location of sample 300 gathers image The image collected is to maintain clearly.

In some embodiments, chasing after Jiao includes step：When camera lens module 104 is in save location, camera lens mould is obtained Group 104 and the relative position of sample 300；When driving the sample 300 to move using microscope carrier 103, the motion of control camera lens module 104 with Keep relative position constant.So, it is ensured that imaging device 102 collects when the diverse location of sample 300 gathers image Image is to maintain clearly, and realization chases after Jiao.

Specifically, due to the physical error of microscope carrier 103 and/or sample 300, may result in sample 300 be it is inclined, because This, is when driving sample 300 to move using microscope carrier 103, the different position in the surface of sample 300 and the distance of camera lens module 104 Minor variations can occur.Therefore, when sample 300 moves relative to the optical axis OP of camera lens module 104, imaging device 102 is to sample 300 image space is always held at clear face position.This process is referred to as chasing after Jiao.

Sample 300 is driven to move using microscope carrier 103, including sample 300 moves along the X1 axles parallel to X-axis, and sample 300 Moved along parallel to the Y1 axles of Y-axis, and plane X1Y1 of the sample 300 along X1 axles and Y1 axis limits is moved, and sample 300 is along inclination Moved in X-axis, and sample 300 moves along Y-axis is favoured, and sample 300 moves along the plane XY for favouring X-axis and Y-axis restriction.

In some embodiments, when driving the movement of sample 300 using microscope carrier 103, the present bit of camera lens module 104 is judged Put and whether exceed the 3rd setting position；When exceeding three setting positions in the current location of camera lens module 104, microscope carrier 103 is utilized Sample 300 is driven to be moved along optical axis OP and carry out focusing step；Working as setting number and camera lens module 104 is reached in mobile number When front position still exceeds three setting positions, judgement chases after burnt failure.In this way, the restriction of the 3rd setting position and mobile number makes mirror Head mould group 104 can be focused again when chasing after burnt failure.

Specifically, in the utility model example, the 3rd setting position can be nPos, and coordinate position is in Z corresponding to nPos On the negative axle of axle, and coordinate position corresponding to nPos is more than coordinate position corresponding to the second setting position farlimit.Work as camera lens Coordinate position corresponding to the current location of module 104 is less than corresponding to the 3rd setting position during coordinate position, judges camera lens module 104 current location exceeds the 3rd setting position.

When exceeding three setting positions in the current location for judging camera lens module 104 first, it can be focused again with right The position of camera lens module 104 is adjusted to attempt to chase after burnt success.During Jiao is chased after, if the number of moving lens module 104 When reaching setting number, the current location of camera lens module 104 still exceeds the 3rd setting position, then can not chase after Jiao, and judgement chases after burnt mistake Lose, suspend and clear face is found in focusing again.

Coordinate position corresponding to 3rd setting position is empirical value, during less than the value, figure that imaging device 102 collects Picture is fuzzy and very maximum probability chases after burnt failure.It is empirical value to set number, can make specific setting according to actual conditions.

In some embodiments, when not exceeding three setting positions in the current location of camera lens module 104, judge relative Position is constant.In some embodiments, relative position includes relative distance and relative direction.Further, it is simplified operation, Relative position can refer to relative distance, and relative position is constant to be referred to, the object distance of the imaging system of imaging device 102 is constant, may be such that The diverse location of sample 300 can be by imaging device blur-free imaging.

In some embodiments, Fig. 3-Fig. 4 and Figure 10-Figure 12 please be join, bogey 100 includes bottom plate 101 and elasticity Support component 40.Microscope carrier 103 is fixed on bottom plate 101 and is provided with storage tank 242, and the bottom of storage tank 242 is provided with through hole 2422. Temperature control device 301 is connected with the sample 300 in storage tank 242 by through hole 2422 and propped up by the elasticity of resilient support assemblies 40 Support is on bottom plate 101.

In the bogey 100 of the utility model embodiment, temperature control device 301 is propped up by the elasticity of resilient support assemblies 40 Support is on bottom plate 101 so that when sample 300 is loaded into storage tank 242, sample 300 and temperature control device 301 and bottom plate 101 Contact as Elastic Contact, damaged when effectively preventing sample 300 from assembling.

Specifically, when microscope carrier 103 does not have device sample 300 also, resilient support assemblies 40 push up temperature control device 301 to accommodating Groove 242, temperature control device 301 is set to be protruded relative to the bottom surface of storage tank 242.When sample 300 is loaded to storage tank 242, pass through Sample 300 is pressed to temperature control device 301 by the frame 221 that clamps of microscope carrier 103, and at the same time, the temperature control device 301 pressed compresses downwards Resilient support assemblies 40, resilient support assemblies 40 is produced elastic force, finally sample 300 is locked on microscope carrier 103.In sample When 300 whole process loaded and sample 300 are locked, sample 300 and temperature control device 301 and bottom plate 101 are Elastic Contact, are had Effect prevent sample 300 assemble and sequencing procedure in damage.

In some embodiments, resilient support assemblies 40 include guiding tube 43 and elastic component 44, and temperature control device 301 includes Temperature control portion 312 and guide posts 341, guide posts 341 are arranged far from the side in temperature control portion 312 of storage tank 242, guiding tube 43 It is fixed on bottom plate 101, guide posts 341 wear elastic component 44 and guiding tube 43, and the elasticity of elastic component 44 is contradicted in the He of temperature control portion 312 Between guiding tube 43.In this way, elastic component 44 can provide elastic force, meanwhile, the cooperation of guiding tube 43 and guide posts 341 also can Ensure that the movement of temperature control device 301 is relatively stable.

It is appreciated that when sample 300 is loaded into storage tank 242, contacts and push with temperature control device 301, now elastic component 44 are compressed, and the reaction force after elastic component 44 is compressed provides sufficient contact for sample 300 and temperature control device 301, with this Meanwhile the Elastic Contact that elastic component 44 provides ensures that sample 300 is less likely to be damaged during pushing.

In some embodiments, elastic component 44 can be spring, so, it is possible to reduce being manufactured into for bogey 100 This.

In some embodiments, elastic component 44 can be the element that rubber cylinder etc. possesses elasticity, not do herein any Limitation.

In some embodiments, guiding tube 43 is linear bearing, and the ball slip of guide posts 341 and linear bearing connects Touch.In this way, linear bearing can also reduce resistance when temperature control device 301 moves while the movement of temperature control device 301 is guided.

It is appreciated that during sample 300 pushes, temperature control device 301 can be press against and be moved down, and in other words, be led Drawing post 341 needs slide downward, and the ball of linear bearing can reduce friction in 341 slide downward of guide posts.

Fig. 4 and Figure 10 are referred to, in some embodiments, temperature control device 301 includes fixed plate 321, temperature conduction plate 322nd, temperature control element 323 and guide posts 341, temperature control element 323 are folded between fixed plate 321 and temperature conduction plate 322, temperature control Element 323 contacts with temperature conduction plate 322 and fixed plate 321, and temperature conduction plate 322 is used for being loaded in storage tank 242 Sample 300 is contacted, and guide posts 341 are arranged far from the surface of fixed plate 321 of temperature control element 323, and guide posts 341 wear bullet Property support component 40.In this way, realizing the function of temperature control device 301 using better simply structure, bogey 100 is reduced Cost.

It is appreciated that fixed plate 321, which is temperature control element 323, provides support, temperature conduction plate 322 produces temperature control element 323 Raw temperature is transferred to sample 300, so as to realize the control of the temperature of sample 300.Guide posts 341 wear resilient support assemblies 40, Using the elastic force of resilient support assemblies 40 sample 300 and temperature control device 301 are in close contact.

In some embodiments, temperature conduction plate 322 and fixed plate 321 can be made up of metal material, for conducting The temperature of temperature control element 323.

In one example, temperature control element 323 is Peltier.Using Peltier effect, temperature control element 323 can realize system Cold heating.For example, the electric current by applying different directions to temperature control element 323 so that the upper surface refrigeration of temperature control element 323, Lower surface heats, or the upper surface heating of temperature control element 323, lower surface refrigeration.With the temperature of the upper surface of temperature control element 323 Conductive plate 322 is spent by the upper surface temperature conduction of temperature control element 323 to sample 300, and then realizes the temperature control to sample 300 System.

In some embodiments, heat-conducting layer, heat-conducting layer can be set between temperature conduction plate 322 and temperature control element 323 By the temperature conduction of temperature control element 323 to temperature conduction plate 322.In this way, improve temperature control element 323 and temperature conduction plate 322 Between pyroconductivity.In one example, heat-conducting layer is layer of silica gel.

In some embodiments, temperature control device 301 also includes the fixed plate 321 for being arranged far from temperature control element 323 Water-bath room 324 on surface, water-bath room 324 are arranged at intervals with guide posts 341.In this way, when temperature control element 323 works, water-bath The heat of temperature control element 323 can be taken away in time by fixed plate 321 room 324.

In some embodiments, temperature control device 301 includes the temperature sensor 325 being arranged on temperature conduction plate 322. It so, it is possible to realize the temperature detection to temperature conduction plate 322, be advantageous to the accurate temperature control to sample 300.

Specifically, when bogey 100 works, because temperature conduction plate 322 and sample 300 are in close contact, temperature conduction The temperature of plate 322 equivalent to sample 300 temperature, so temperature sensor 325 can obtain the temperature of sample 300 and feed back to External control device, so as to which external control device can be according to the temperature of the feedback control sample 300 of temperature sensor 325.

In the utility model embodiment, temperature conduction plate 322 can offer accommodating hole, and temperature sensor 325 is worn In accommodating hole, the temperature of sample 300 so can be more accurately monitored.In a detailed embodiment, bogey 100 can It is ± 0.1 DEG C or ± 0.5 DEG C to realize the surface temperature degree of accuracy of sample 300；The surface temperature fluctuation degree of sample 300 is not more than 0.1 DEG C Or 0.5 DEG C；It is not more than 1min from 25 DEG C to the 65 DEG C heating-up times of room temperature, 25 DEG C of temperature fall times of room temperature is down to from 65 DEG C and are not more than 1.5min.The temperature control of sample 300 is accurate, can lift biochemical reaction efficiency, reduces the testing time.

Figure 11 is referred to, in some embodiments, water-bath room 324 includes heat sink 3242, cover plate 3244, liquid inlet joint 3246 and go out liquid joint 3248.Flow path groove is provided with heat sink 3242, heat sink 3242 contacts with fixed plate 321.Cover plate 3244 are connected with heat sink 3242 and cover flow path groove and form sap cavity, and sap cavity is used to accommodate coolant, the company of setting on cover plate 3244 The inlet and liquid outlet of fluid-through chamber.Liquid inlet joint 3246 connects inlet.Go out liquid joint 3248 and connect liquid outlet.It is in this way, real The efficient radiator structure of water-bath room 324 is showed.

It is appreciated that water-bath room 324 is used to provide coolant circulation, it is the temperature contacted with fixed plate 321 using coolant Control the lower surface radiating of element 323.The flow path groove set on heat sink 3242 can increase heat sink 3242 and fixed plate 321 Contact area, so as to improve the efficiency of radiating.

In some embodiments, coolant can be water.It so, it is possible the cost of reduction bogey 100.

In some embodiments, coolant can be the coolant specially made, not do any restrictions herein.Special system The coolant of work can ensure that the capacity of heat transmission reaches ideal state.

Referring to Figure 11, in some embodiments, temperature control device 301 includes temperature controlled water bath device 326, temperature control water Bath apparatus 326 includes fin 3262, infusion pump 3264 and cooling device 3266.Fin 3262 is offered for supplying coolant The stream flowed through, the import of stream connect out liquid joint 3248.Infusion pump 3264 connects outlet and the liquid inlet joint 3246 of stream. Cooling device 3266 is used for cold sink 3262.It so, it is possible to take the heat of water-bath room 324 to temperature controlled water bath device 326 And dissipated by fin 3262.

Specifically, temperature controlled water bath device 326 is used for the heat exchange for accelerating coolant and external environment condition, so that it is guaranteed that coolant Fast cooling.Fin 3262 is used to make coolant and external environment condition carry out heat exchange, and infusion pump 3264 is used to promote to cool down The circulation of liquid, cooling device 3266 are used for the heat exchange for accelerating the coolant in fin 3262.

In some embodiments, cooling device 3266 can be fan.Dried, increased to fin 3262 by fan The convection current of air, so as to realize the heat exchange for accelerating the coolant in fin 3262.

In some embodiments, temperature control device 301 includes thermal insulation barriers 327.Thermal insulation barriers 327 are arranged on temperature conduction plate Between 322 and fixed plate 321.It so, it is possible to avoid temperature conduction plate 322 from by the temperature interference of fixed plate 321 being caused to sample The temperature control of product 300 is inaccurate.

Specifically, temperature conduction plate 322 and fixed plate 321 contact with two different faces of temperature control element 323 respectively, two The temperature in face caused temperature in the course of work of temperature control element 323 is different.In order to ensure that temperature conduction plate 322 exactly will The temperature conduction of temperature control element 323 is to sample 300, so utilizing the blocking temperature conductive plate 322 of thermal insulation barriers 327 and fixed plate 321 Between temperature transmission, so that it is guaranteed that the temperature of temperature conduction plate 322 is not influenceed by the temperature of fixed plate 321.

In some embodiments, thermal insulation barriers 327 can be heat insulation foam.Heat insulation foam can be kept away while realizing heat-insulated Exempt from the temperature control element 323 damaged between temperature conduction plate 322 and fixed plate 321.

In some embodiments, temperature control device 301 includes being used for the surface 328 contacted with sample 300, and surface 328 is The black surface of matt.In this way, the laser reflection sent when can prevent bogey 100 from working comes to the print imaging belt of sample 300 Negative effect.

Specifically, the runner in sample 300 is transparent, when bogey 100 works, need to launch laser to sample 300, To excite the sample in sample 300 to send fluorescence, and the image of sample is formed by gathering fluorescence.By will be with sample 300 The surface 328 of contact is arranged to the black surface of matt, effectively reduces the reflection of laser.

In some embodiments, surface 328 is the upper surface of the temperature conduction plate 322 contacted with sample 300.

In some embodiments, microscope carrier 103 includes clamping frame 221 and support base 241, clamps frame 221 and rotationally connects It is connected on support base 241, support base 241 offers storage tank 242, and support base 241 is arranged on bottom plate 101.In this way, pass through folder Frame up 221 rotations relative to support base 241, can facilitate loading and the unloading of sample 300.

Specifically, when sample 300 is placed into storage tank 242, connected using the rotation for clamping frame 221 and support base 241 Connect to frame up with rotating clip and 221 sample 300 is pressed into temperature control device 301, so that it is guaranteed that sample 300 and temperature control device 301 fill Tap is touched.Afterwards, clamp frame 221 and be locked at support base 241 for example, by the mode of buckle, to compress sample 300, ensure sample 300 stability in sequencing procedure.The junction for clamping frame 221 and support base 241 may be provided with torsion spring, clamp frame 221 During unblock, torsion spring, which can drive, to be clamped frame 221 and is opened relative to support base 241.

Figure 12 is referred to, in some embodiments, button 246, the connection buckle of button 246 are provided with support base 241 248.In this way, the motion state of buckle 248 can be controlled by button 246, the locking for clamping frame 221 and support base 241 is released State, unlocked so as to which frame 221 will be clamped.

Specifically, when button 246 is pressed, button 246 can drive buckle 248 remote by way of lever connection Clamp frame 221 to move, so as to unlock the locking state for clamping frame 221 and buckle 248, it is possible to achieve the assembling of sample 300 is torn open Unload.When button 246 is released, buckle 248 loses external force effect, and buckle 248 resets.When clamping the closing of frame 221, frame is clamped 221 above lock with the card of buckle 248 again and clamp frame 221.

Referring to Figure 12, in some embodiments, support base 241 includes panel 244, is offered on panel 244 Button pass through hole 2442, button 246 wear button pass through hole 2442, and the bottom of button 246 is convexly equipped with flange 2462, flange 2462 and face The lower surface of plate 244 contradicts.

In this way, panel 244 can press flange 2462, so that button 246 is pressed and resetted smoothly, and can make Obtain when button 246 resets and limited, will not be run out of.

In some embodiments, Figure 13, Fig. 3, Fig. 4 and Fig. 6 please be join, the utility model embodiment provides a kind of control The method of sequencing reaction, sequencing reaction is controlled using Sequence Detection System, Sequence Detection System includes light Detecting system is learned, Systems for optical inspection includes imaging device 102 and bogey 100, and bogey 100 includes temperature control device 301 and microscope carrier 103, imaging device 102 includes camera lens module 104, and camera lens module 104 includes optical axis OP, and microscope carrier 103 is used to carry Sample, temperature control device 301 are used for the temperature for adjusting sample.Sequencing reaction is carried out using imaging device 102 to sample IMAQ.The method of control sequence measure reaction includes：Using Sequence Detection System carry out sequencing reaction before or Person sets the model for the temperature fluctuation for allowing sample using temperature control device 301 when carrying out sequencing using Sequence Detection System Enclose, so that positional fluctuation scope of the camera lens module 104 along optical axis OP is located in preset range.It is anti-using above-mentioned control sequence measure The method answered, it can be reduced or avoided by the positional fluctuation scope control of camera lens module 104 within a preset range to imaging device 102 carry out adverse effect during IMAQ.

In some embodiments, Systems for optical inspection is preset with the scope and camera lens module of the temperature fluctuation of sample 300 104 along the positional fluctuation scope of optical axis corresponding relation, according to corresponding relation, control camera lens module 104 is along optical axis OP position Fluctuation range is located in preset range.In this way, the method for control sequence measure reaction can be obtained the edge of camera lens module 104 rapidly The scope of the positional fluctuation scope control of the optical axis temperature fluctuation of corresponding sample 300 within a preset range.

Specifically, the scope of the temperature fluctuation of sample 300 is corresponding with positional fluctuation scope of the camera lens module 104 along optical axis Relation is storable in Systems for optical inspection.In one example, alleged corresponding relation can join table 1.In some embodiments In, imaging device 102 includes focusing module 106, and IMAQ is carried out to sample 300 using imaging device 102, including：Utilize Focusing module 106 and camera lens module 104 are focused to sample 300.

It is pointed out that it is above-mentioned in Systems for optical inspection on the embodiment of focusing and explaining for beneficial effect The bright focusing being also applied in the Sequence Detection System of present embodiment, to avoid redundancy, is no longer developed in details herein.

In some embodiments, Figure 14 please be join, sequencing reaction includes the first biochemical reaction and the second biochemical reaction, First biochemical reaction and the second biochemical reaction are carried out on reaction unit 200, and Sequence Detection System includes fluid means 500, stream The coupled reaction device 200 of body device 500, reaction unit 200 include first module 41 and second unit 42, and sample 300 is placed in On one unit 41 and second unit 42, it is second biochemical anti-that defined nucleotide sequence measure, which reacts the one kind included to repeat unit S112, Answer the-the first biochemical reaction-IMAQ, the method for control sequence measure reaction be included in complete following initial step S111 it Afterwards so that when the second biochemistry for making a progress sample 300 in first module 41 and second unit 42 using fluid means 500 While reaction and the first biochemical reaction, IMAQ is carried out to the sample 300 of another unit using imaging device 102, just Beginning step S111 includes step：A makes the sample on one in first module 41 and second unit 42 using fluid means 500 300 carry out the first biochemical reaction, and b is carried out using imaging device 102 to the sample 300 on the unit after carrying out the first biochemical reaction IMAQ, c made using fluid means 500 another in first module 41 and second unit 42 on sample 300 carry out the One biochemical reaction.The method of above-mentioned control sequence measure reaction, in being reacted based on sequencing, reaction unit 200 is at least divided For two units, one of unit is set to utilize imaging device 102 while biochemical reaction is carried out using fluid means 500 It is IMAQ to carry out IMAQ to another unit, and then can reduce the time used in sequencing, improves sequence survey Determine efficiency.

Specifically, in the embodiment of the method for the control sequence measure reaction shown in Figure 14, sample 300 is to treat test sample Product, sample 300 are located on reaction unit 200 (supporting arrangement).In the utility model embodiment, sample can will be also carried 300 reaction unit 200 is placed on bogey 100 to carry out sequencing reaction.

The explanation of the embodiment and beneficial effect of bogey 100 in the method for control sequence measure reaction can Join the embodiment of the bogey 100 in above-mentioned Systems for optical inspection, to avoid redundancy, be no longer developed in details herein.

The time difference of sequencing reaction mesophytization reaction and information of the inventor based on discovery, based on reaction unit and The number of imaging device in Sequence Detection System, reaction unit is at least divided into two units, the computer made as above can be held All or part of method for carrying out sequencing reaction of capable parallel control calling device/system, makes full use of sequencing reaction In key step time difference, greatly improve sequencing reaction efficiency.

Usually, the device/system needed for sequencing reaction, from the point of view of hardware spending, imaging device/system are carried out The cost more than fluid means/system is spent, the cost of fluid means/system is more than the cost of reaction unit/chip.Utilize this Utility model this method control sequence measure reaction, can make full use of imaging device/system, fluid means/system and Reaction unit, further reduce sequencing cost.

Specifically, in some embodiments, reaction unit 200 can be chip, the He of first module 41 of reaction unit 200 Second unit 42 includes plurality of passages (channel), after initial step S111, the passage and second unit of first module 41 42 passage is staggering, nonsynchronous, is mutually independent of each other in sequencing reaction.For example, needing to first module 41 On sample carry out biochemical reaction when, fluid means 500 can to first module 41 convey reaction reagent, now, will not make Identical reagent enters second unit 42, and vice versa.

In one example, core is carried out in the single-molecule sequencing platform using total internal reflection (TIRF) optical system detection Acid sequence determines, the experience of the ratio based on the data volume needed for the analysis of follow-up hereditary information and the valid data after processing Value, it is about 300 visuals field (FOV) to estimate IMAQ number corresponding to required original data volume.Sequencing is taken turns one In reaction, using imaging device 102 control time needed for 300 FOV of mobile response device 200 and collection substantially with utilization The summation that fluid means 500 carries out the time of the first biochemical reaction and the second biochemical reaction is equal, using it is of the present utility model this The method of one embodiment, it is possible to increase one times of reaction efficiency.

It will be understood by those skilled in the art that if in some other situations, the data volume as needed for hereditary information analysis subtracts The ratio of valid data less and/or after processing improves so that the FOV numbers of collection tail off needed for each round sequencing reaction, that is, scheme Reduced the time required to picture collection or the total time of biochemical reaction is elongated, react dress by m using method of the present utility model Put and be divided into n unit, m, n are the integer more than or equal to 1, and n is more than or equal to twice of m, each unit is in a wheel/no Different step or stage with wheel sequencing reaction, can make full use of imaging device 102 and fluid means 500, improve reaction effect Rate.If those skilled in the art are further appreciated that in the case of some are opposite with above example, the time required to biochemical reaction Tail off, can also make full use of the number of unit on reaction unit 200 to configure to carry using this method of the present utility model High efficiency.

In some embodiments, before sequencing reaction is carried out, the first module 41 and second of reaction unit 200 is single The fixed sample for needing sequencing, the sample for treating sequencing are, for example, to have double-strand or list on the surface of the passage of member 42 The DNA of chain structure.

In the utility model embodiment, it is second the-the first biochemical reaction of biochemical reaction-figure to repeat unit S112 As collection, refer to when to some unit progress sequencing reaction of reaction unit 200, successively to the sample on the unit Product carry out the second biochemical reaction, the first biochemical reaction and IMAQ.When repeating unit and being performed a number of times, this practicality is new The method of type embodiment occurs carries out the first biochemical reaction-biochemical reaction of IMAQ-the second to the sample on unit Process is repeated, and/or occurs carrying out the-the first biochemical reaction of biochemical reaction of IMAQ-the second to the sample on unit Repeat process.It should be noted that usually, alleged sequencing reaction is often through less than one circulation：First is biochemical anti- Should, IMAQ and the second biochemical reaction, it becomes possible to complete the measure of an at least one base, alleged base be selected from A, T, C, At least one of G and U.It will be understood by those skilled in the art that the definition to " repeating unit " in the utility model, The utility model merely for convenience of description, be not limited the reaction sequence determined in sequencing reaction.

In the utility model embodiment, second is carried out when making the sample in first module 41 using fluid means 500 While biochemical reaction and the first biochemical reaction, IMAQ is carried out to the sample on second unit 42 using imaging device 102, Then, according to unit is repeated, the second biochemical reaction has been carried out making the sample in first module 41 using fluid means 500 After the first biochemical reaction, while carrying out IMAQ to the sample in first module 41 using imaging device 102, to the After sample on two units 42 carries out IMAQ, the sample on second unit 42 is set to carry out the second life using fluid means 500 Change reaction and the first biochemical reaction.

It is biochemical when making the sample on second unit 42 carry out second using fluid means 500 in other embodiment While reaction and the first biochemical reaction, IMAQ is carried out to the sample in first module 41 using imaging device 102, so Afterwards, according to repeating unit, made using fluid means 500 sample on second unit 42 carried out the second biochemical reaction and After first biochemical reaction, while carrying out IMAQ to the sample on second unit 42 using imaging device 102, to first After sample on unit 41 carries out IMAQ, the sample in first module 41 is set to carry out second using fluid means 500 biochemical Reaction and the first biochemical reaction.

In the utility model embodiment, Figure 14 please be join, in initial step S111, a makes using fluid means 500 Sample on one unit 41 carries out the first biochemical reaction；B is single to first after carrying out the first biochemical reaction using imaging device 102 Sample in member 41 carries out IMAQ；It is biochemical anti-that c makes the sample on second unit 42 carry out first using fluid means 500 Should.

In another embodiment, in the initial step, a enters the sample on second unit 42 using fluid means 500 The biochemical reaction of row first；B carries out figure using imaging device 102 to the sample on the second unit 42 after carrying out the first biochemical reaction As collection；C makes the sample in first module 41 carry out the first biochemical reaction using fluid means 500.

IMAQ is carried out to sample using imaging device 102 and forms view data, view data may be output to sequence survey Determine the other device/modules of system to be handled to obtain corresponding image.

In some embodiments, step a and step c are carried out simultaneously, or step b and step c are carried out simultaneously, or step b Carried out before step c, or step b is carried out after step c.In this way, the implementation tool for the method being controlled to sequencing There are more flexibilities.

Specifically, in the utility model embodiment, in step a, made using fluid means 500 in first module 41 Sample when carrying out the first biochemical reaction, sample on second unit 42 is first biochemical anti-not by the sample in first module 41 It should influence.Vice versa.

It is preferred that step b and step c are carried out simultaneously, the efficiency of method is so further improved.

In some embodiments, the first biochemical reaction includes extension, and the second biochemical reaction is cut off including group.Such as This, makes wider to the method application that is controlled of sequencing reaction.

Specifically, in some embodiments, before being carried out to sequencing reaction, the first of reaction unit 200 is single The sample for needing sequencing, i.e. template strand are fixed in the passage of member 41 and second unit 42.Polymerase/ligase extension is anti- Should be connected to based on base complementrity, by specific substrates on the sample for treating sequencing, and be examined using what is carried on substrate Group is surveyed to determine the type with reference to upper substrate, to determine sequence.In one example, can detect group includes fluorescent base Group, fluorescence can be sent under the laser of specific wavelength.

Group excision (cleave) reaction is carried on the substrate being incorporated on the sample (template) for treating sequencing Group is cut off, so that next base of template can continue to be measured, i.e. first module 41 and/or second unit 42 On sample can continue to carry out sequencing reaction.

In some embodiments, extension includes being sequenced in connection and is sequenced in synthesis.

In some embodiments, the second biochemical reaction includes capping.

Alleged caps the group/key being exposed after predominantly blocking group excision.In one example, first is biochemical Reaction includes base extension, and the structure of added substrate terminates group-connection unit-luminophore for A/T/C/G-, wherein Termination group for light and/or chemistry can cleavable groups, substrate is carried luminophore by connection unit (linker).Second Biochemical reaction includes group excision, can be after cleavable groups by light and/or chemical ablation, and the group being exposed be sulfydryl, logical Cross and cap such as by adding alkylating reagent, sulfydryl can be protected not oxidized.So so that sequencing reaction is controlled The method application of system is wider.

In some embodiments, IMAQ also includes adding imaging agents.Alleged imaging agents include it is oxidation resistant into Point, such as watermiscible vitamin E (Trolox) etc., it can avoid or reduce light in image acquisition process and be damaged to caused by sample Wound influences.

It is preferred that the light that laser excitation sample is sent is fluorescence, ambient light can be reduced imaging device is adopted to sample image Adverse effect during collection.

Further illustrate：" signal collection " process includes plus imaging agents, IMAQ are (real in the utility model Apply in mode, imaging agents will be added to be placed in IMAQ)；After cleave, buffer solution (buffer1) cleaning, cap and (add certain It is protect reagent, relevant with substrate structure), buffer2 cleanings (buffer1,2 can be with identical, also can be different) again.

In some embodiments, Figure 13 please be join, fluid means 500 includes valve component 10 and drive component 50, driving Component 50 connects valve component 10 by reaction unit 200, makes first module 41 and/or second single using fluid means 500 When sample in member 42 carries out the first biochemical reaction and/or the second biochemical reaction, it is different that valve component 10 is used for switching connection Reagent, drive component 50 make valve component 10 export reagent to first module 41 and/or second unit 42.In this way, pass through valve body Component 10 and drive component 50, can easily by sequencing react needed for different reagents input to first module 41 and/ Or second unit 42.

Specifically, in the utility model embodiment, fluid means 500 includes reagent component, and reagent includes the first examination Agent, the second reagent and the 3rd reagent, reagent component include containing the first reagent bottle 11 of the first reagent, contain the of the second reagent Two reagent bottles 12 and the 3rd reagent bottle 13 for containing the 3rd reagent, valve component 10 connect the first reagent bottle 11, the by pipeline Two reagent bottles 12 and the 3rd reagent bottle 13.The switching of valve component 10 connects different reagent bottle, so that drive component 50 can be from Reagent is extracted in the reagent bottle connected with valve component 10 to first module 41 and/or second unit 42.

In some embodiments, valve component 10 includes the first multiple-way valve 20 and the first triple valve 30, the first multiple-way valve 20 switchings connect different reagent to the first triple valve 30, the first triple valve 30 by the reagent that the first multiple-way valve 20 exports export to First module 41 and/or second unit 42.In this way, drive component 50 is realized by the first multiple-way valve 20 and the first triple valve 30 Valve component 10 is set to export different reagents to first module 41 and/or second unit 42.

Specifically, in the utility model embodiment, the pipeline of the first multiple-way valve 20 connects the examination of the first reagent bottle 11, second Agent bottle 12, the 3rd reagent bottle 13 and the first triple valve 30, the first multiple-way valve 20 are used for the first reagent bottle 11, the second reagent bottle 12 Or the 3rd reagent bottle 13 be connected with the first triple valve 30.The pipeline of first triple valve 30 connection first module 41, second unit 42 And first multiple-way valve 20, the first triple valve 30 are used to first module 41 or second unit 42 with the first multiple-way valve 20 being connected.

In some embodiments, the first reagent is sequencing reagent, and the second reagent is that group cuts off reagent, and the 3rd reagent is Imaging agents, the first multiple-way valve 20 include the first extraction opening 21 of the first reagent bottle 11 of connection, the of the second reagent bottle 12 of connection Two extraction openings 22 and the 3rd extraction opening 23 for being connected to the 3rd reagent bottle 13, and a liquid outlet 24.Liquid outlet 24 and first The extraction opening 22 of extraction opening 21 or second or the 3rd extraction opening 23 are connected.Sequencing reagent is comprising extension at least a portion The reagent of reactant, the reagent e.g. comprising substrate and polymerase/ligase.Substrate carries detectable group, such as with glimmering Light group.

First triple valve 30 includes liquid sucting port 31, the first diffluence pass 32 and the second diffluence pass 33, the connection of liquid sucting port 31 first The diffluence pass 33 of diffluence pass 32 or second.Liquid sucting port 31 connects liquid outlet 24.First module 41 and second unit 42 are respectively communicated with One diffluence pass 32 and the second diffluence pass 33.

In the utility model embodiment, the first multiple-way valve 20 is rotary valve, the first extraction opening 21, the second extraction opening 22 And the 3rd extraction opening 23 respectively around around liquid outlet 24, the first extraction opening 21, the second extraction opening 22 and the 3rd extraction opening 23 Liquid outlet 24 is connected by the rotating pipe 25 rotated around liquid outlet 24.Rotating pipe 25 can rotate to the first extraction opening successively 21st, the position of the second extraction opening 22 and the 3rd extraction opening 23, so as to realize liquid outlet 24 can be sequentially communicated the first reagent bottle 11, Second reagent bottle 12 and the 3rd reagent bottle 13, i.e. reaction unit 200 can be respectively from the first reagent bottle 11, the and of the second reagent bottle 12 3rd reagent bottle 13 obtains different reagents, and then sample is carried out the first biochemical reaction, the second biochemical reaction and IMAQ. In other embodiments, the connection order of the extraction opening 21 of liquid outlet 24 and first, the second extraction opening 22 and the 3rd extraction opening 23 It can not also limit.

In some embodiments, when the liquid sucting port 31 of the first triple valve 30 is connected with the first diffluence pass 32, liquid sucting port 31 disconnect with the second diffluence pass 33, and vice versa.Liquid sucting port 31 can connect the first diffluence pass according to needed for sequencing 32 or second diffluence pass 33, i.e., when the sample in first module 41 carries out the second biochemical reaction and the first biochemical reaction, first Diffluence pass 32 is connected with liquid sucting port 30, so as to which liquid sucting port 30 provides required second through the first diffluence pass 32 to first module 41 Reagent and the first reagent, after first module 41 has obtained the second reagent and the first reagent, the second diffluence pass 33 and liquid sucting port 31 It is connected, so that second unit 42 obtains the 3rd reagent, imaging device 102 can carry out figure to the sample on second unit 42 As collection.

After the completion for the treatment of the sample image collection on second unit 42, second unit 42 starts to obtain the second examination through liquid sucting port 31 Agent and the first reagent, the sample on second unit 42 is set to carry out the second biochemical reaction and the first biochemical reaction, in second unit 42 After having obtained the second reagent and the first reagent, the first diffluence pass 32 is connected with liquid sucting port 31, and first module 41 obtains the 3rd examination Agent, imaging device 102 can carry out IMAQ to the sample in first module 41, so as to effectively reduce the time of sequencing, Improve the efficiency of sequencing.

In some embodiments, drive component 50 includes the first pump 51 and the second pump 52, and the first pump 51 is single by first Member 41 connects valve component 10, and the second pump 52 connects valve component 10 by second unit 42, makes the using fluid means 500 When sample on one unit 41 and/or second unit 42 carries out the first biochemical reaction and/or the second biochemical reaction, the first pump is utilized 51 make valve component 10 export reagent to first module 41, and/using the second pump 52 valve component 10 is exported reagent to second Unit 42.In this way, it can be realized the reagent delivery that valve component 10 exports to first respectively using the first pump 51 and the second pump 52 Unit 41 and/or second unit 42, convenient operation.

Specifically, the first pump 51 connects first module 41 and second unit 42 with the second pump 52 difference pipeline.

In the utility model example, the first pump 51 connects the first diffluence pass of the first triple valve by first module 41, Second pump 52 connects the second diffluence pass of the first triple valve by second unit 42, and during work, the first pump 51 is to first module 41 Negative pressure is provided, so that first module 41 obtains the second reagent successively and the first reagent carries out the second biochemical reaction and the first biochemistry is anti- Should, after first module 41 has obtained the second reagent and the first reagent, the first pump 51 stops providing negative pressure, and the second pump 52 provides negative Press so that second unit 42 obtains the 3rd reagent, and carries out image to the sample on second unit 42 using imaging device 102 and adopt Collection.

It is pointed out that when the sample on making first module 41 carries out the second biochemical reaction and the first biochemical reaction, Liquid outlet 24 is sequentially communicated the second extraction opening 22 and the first extraction opening 21 to extract the second reagent and the first reagent, liquid sucting port 31 with First diffluence pass 32 connects, when the first pump 51 is to the offer negative pressure of first module 41 so that the second reagent and the first reagent enter successively In the passage for entering first module 41.

After first module 41 has obtained the second reagent and the first reagent, the first pump 51 stops providing negative pressure, liquid outlet 24 The 3rd extraction opening 23 is connected to extract the 3rd reagent, liquid sucting port 24 connects with the second diffluence pass 33, and the second pump 52 is to second unit 42 provide negative pressure so that and the 3rd reagent enters in the passage of second unit 42, and using imaging device 102 on second unit 42 Sample carry out IMAQ.Therefore, valve component 10, drive component 50 and imaging device 102 cooperate, and make the first list Sample in member 41 carries out figure while the second biochemical reaction and the first biochemical reaction is carried out to the sample on second unit 42 As collection.Vice versa.

In some embodiments, fluid means 500 includes at least one first container and sequencing reagent configuration component 60, Reagent includes sequencing reagent, and first is carried out making the sample on first module 41 and/or second unit 42 using fluid means 500 When biochemical reaction and/or the second biochemical reaction, sequencing reagent configuration component 60, which exports sequencing reagent to valve component 10, to be connected In the first logical container.In this way, convenient be added to first module 41 and second unit by the reagent for carrying out sequencing reaction In 42.

Specifically, in the utility model example, the first container is the first reagent bottle 11.In one example, first holds The number of device is multiple.

Sequencing reagent configuration component 60 includes multiple sequencing reagent stock bottles 61, the second multiple-way valve 62, the second triple valve 63 With the 3rd pump 64.Multiple sequencing reagent stock bottles 61 are to contain a variety of sequencing reagent raw materials, the second multiple-way valve 62 pipeline simultaneously Multiple sequencing reagent stock bottles 61 are connected, and pipeline connects the second triple valve 63.Second triple valve 63 goes back pipeline connection the 3rd The reagent bottle 11 of pump 64 and first.3rd pump 64 is through the second triple valve 63 and the second multiple-way valve 62 and one of sequencing reagent raw material Bottle 61 is connected.First reagent bottle 11 is connected through the second triple valve 63 with the 3rd pump 64.Wherein, the 3rd pump 64 successively with it is multiple Sequencing reagent stock bottle 61 is connected, to extract the sequencing reagent raw material in multiple sequencing reagent stock bottles 61, and mixed configuration Into sequencing reagent, the 3rd pump 64 is connected with the first reagent bottle 11, for sequencing reagent to be injected in the first reagent bottle 11.

In present embodiment, multiple sequencing reagent stock bottles 61 contain different sequencing reagent raw materials respectively, so as to The sequencing reagent raw material in multiple sequencing reagent stock bottles 61 is extracted successively using the 3rd pump 64, so as to be mixedly configured into sequencing examination Agent.

In one example, the number of sequencing reagent stock bottle 61 is nine, and it is similar to contain different types of nucleosides respectively Solution, archaeal dna polymerase solution and the part of various cushioning liquid or sulfhydryl protected solution of thing (substrate).Multiple sequencings Reagent stock bottle 61 can be disposed on rack for test tube, to be consolidated to multiple sequencing reagent stock bottles 61, while may be used also With the different labels that are sticked respectively to six sequencing reagent stock bottles 61, carry out sequencing reagent raw material next time to facilitate and supplemented, Avoid the cross-infection of sequencing reagent raw material.In other embodiments, the number of sequencing reagent stock bottle 61 can also be two Individual, three, four, five, six, other quantity such as seven or eight, can be according to being actually needed and the characteristic of each solution has Body adjusts.

The setting identical with the structure of the first multiple-way valve 20 of second multiple-way valve 62.Unlike, the second multiple-way valve 62 realizes the Three pumps 64 are connected with multiple sequencing reagent stock bottles 61 successively, and the second multiple-way valve 62 selectes one of sequencing reagent stock bottle 61 connections, and duration is connected by controlling, so as to control the 3rd pump 64 to take out sequencing reagent raw material in sequencing reagent stock bottle 61 The regulation of taken amount.So as to realize that the sequencing reagent raw material of multiple sequencing reagent stock bottles 61 can be configured in proportion, with symbol Close sequencing demand.

The setting identical with the structure of the first triple valve 30 of second triple valve 63.Second triple valve 63 can realize the He of the 3rd pump 64 Second multiple-way valve 62 is connected, and the sequencing reagent raw material in multiple sequencing reagent stock bottles 61 can be extracted so as to the 3rd pump 64, is matched somebody with somebody It is set to sequencing reagent.Second triple valve 63 can realize that the 3rd pump 64 connects with the first reagent bottle 11, so as to which the 3rd pump 64 can be with The sequencing reagent being configured is injected into the first reagent bottle 11.

3rd pump 64 can be provided to multiple sequencing reagent stock bottles 61 through the second triple valve 63 and the second multiple-way valve 62 and born Pressure, to extract the sequencing reagent in multiple sequencing reagent stock bottles 61.3rd pump 64 can also be through the second triple valve 63 to first Malleation is provided in reagent bottle 11, to inject in sequencing reagent to the first reagent bottle 11.

Further, the first blender 65, the first blender are connected between the second triple valve 63 and the first reagent bottle 11 65 set multiple first serpentine tubes 651, and multiple first serpentine tubes 651 join end to end, and are communicated in the He of the second triple valve 63 Between first reagent bottle 11.

In the utility model embodiment, multiple first serpentine tubes 651 are fixed in a fixed plate, and first wriggles Pipeline 651 is S-shaped, and multiple serpentine tubes 651 can be multiple rows of arranged side by side, and be interconnected between often arranging.Wriggled using multiple first Yan pipelines 651 are communicated between the second triple valve 63 and the first reagent bottle 11 so that the sequencing reagent injected from the 3rd pump 64 Enter row buffering by multiple serpentine tubes 651, and increase the flowing stroke of sequencing reagent, so that more in sequencing reagent Kind sequencing reagent raw material is sufficiently mixed, and lifts sequencing reagent reaction efficiency.In other embodiments, multiple serpentine tubes 651 can also be and spiral successively.

The number of first reagent bottle 11 can be one or multiple.In one example, first reagent bottle 11 Number is multiple, and the solution comprising different types of substrate is held respectively.It is logical that sequencing reagent configuration component 60 also includes more than the 3rd Valve 66, pipeline connects multiple first reagent bottles 11, and the second triple valve 63 to the 3rd multiple-way valve 66 simultaneously, and the 3rd pump 64 is through second The multiple-way valve 66 of triple valve 63 and the 3rd is connected with one of them first reagent bottle 11.

In the utility model embodiment, the sequencing reagent in multiple first reagent bottles 11 is different, the first reagent bottle 11 Number be four.It is different that the sequencing reagent source chemicals ratio of multiple sequencing reagent stock bottles 61 is extracted according to the 3rd pump 64, Different sequencing reagents is can be configured to, so as to contain a variety of different sequencing reagents using multiple first reagent bottles 11. The setting identical with the structure of the second multiple-way valve 62 of the structure of 3rd multiple-way valve 66.3rd multiple-way valve 66 can realize the 3rd pump 64 successively to Different sequencing reagents is injected respectively in multiple first reagent bottles 11.Specifically, the 3rd pump 64 is often configured after completing sequencing reagent, First reagent bottle 11 is selected through the second triple valve 63 and the 3rd multiple-way valve 66, and sequencing reagent is injected to the first reagent bottle In 11.In other embodiments, the number of the first reagent bottle 11 can also be two, three, four, five, six or seven The quantity such as individual, can be according to being actually needed and the characteristic of each solution specifically adjusts.

Further, sequencing reagent configuration component 60 also includes the waste liquid bottle 68 of wash reagent bottle 67 and first, wash reagent Bottle 67 is used to contain wash reagent, and wash reagent bottle 67 is connected through the second multiple-way valve 62 and the second triple valve 63 with the 3rd pump 64 Logical, the first waste liquid bottle 68 is used to contain waste liquid, and the first waste liquid bottle 68 is through the 3rd multiple-way valve 66 and the second triple valve 63 and the 3rd pump 64 are connected.

When wash reagent bottle 67 is connected through the second multiple-way valve 62 and the second triple valve 63 with the 3rd pump 64, the 3rd pump 64 can To extract wash reagent in wash reagent bottle 67, to be rinsed to the 3rd pump 64, i.e. the 3rd pump 64 completes a kind of survey in configuration After sequence reagent, before configuring sequencing reagent next time, the wash reagent in wash reagent bottle 67 can be first extracted, after being rinsed Sequencing reagent is configured again, cross-infection occurs so as to avoid configuring two kinds of different gene sequencing.First waste liquid bottle 68 is through When three multiple-way valves 66 and the second triple valve 63 are connected with the 3rd pump 64, the 3rd pump 64 can note the waste liquid for having completed cleaning It is incident upon in the first waste liquid bottle 68, so as to reach the effect of environmental protection recovery.

In the utility model embodiment, sequencing reagent configuration component 60 realizes the on-line mixing of fluid means 500 Function.It is appreciated that in some embodiments, fluid means can also not have on-line mixing function, correspondingly, will can be sequenced Reagent configuration component 60 omits, and equally disclosure satisfy that the fluid path demand and control sequencing reaction fluid path of sequencing reaction.Such energy Enough make that the pipeline of fluid means is simpler, Sequence Detection System more compact.

In some embodiments, fluid means 500 includes second container and imaging agents configuration component 70, and reagent includes Imaging agents, when carrying out IMAQ to the sample in first module 41 and/or second unit 42 using imaging device 102, Imaging agents configuration component 70 exports imaging agents into the second container connected with valve component 10.In this way, convenient will enter The reagent of row sequencing reaction is added in first module 41 and second unit 42.

Specifically, in the utility model example, second container is the 3rd reagent bottle 13.

In the utility model embodiment, imaging agents configuration component 70 includes multiple imaging agents stock bottles 71, the Four multiple-way valves 72, the 3rd triple valve 73 and the 4th pump 74.Multiple imaging agents stock bottles 71 are former to contain a variety of imaging agents Material.Pipeline connects multiple imaging agents stock bottles 71 to 4th multiple-way valve 72 simultaneously, and pipeline connects the 3rd triple valve 73.3rd Triple valve 73 goes back pipeline and connects the 4th pump 74 and the 3rd reagent bottle 13.4th pump 74 is through the 3rd triple valve 73 and the 4th multiple-way valve 72 It is connected with one of imaging agents stock bottle 71.3rd reagent bottle 13 is connected through the 3rd triple valve 73 with the 4th pump 74, Wherein, the 4th pump 74 is connected with multiple imaging agents stock bottles 71 successively, to extract in multiple imaging agents stock bottles 71 Imaging agents raw material, and it is mixedly configured into imaging agents.4th pump 74 is connected with the 3rd reagent bottle 13, for by imaging agents It is injected in the 3rd reagent bottle 13.

In the utility model embodiment, it is former that multiple imaging agents stock bottles 71 contain different imaging agents respectively Material, so as to extract the imaging agents raw material in multiple imaging agents stock bottles 71 successively using the 4th pump 74, so as to mix It is configured to imaging agents.Specifically, the number of imaging agents stock bottle 71 is five.Multiple imaging agents stock bottles 71 can be It is positioned on rack for test tube, to be consolidated to multiple imaging agents stock bottles 71, while can also be respectively to five imaging examinations Agent stock bottle 71 is sticked different labels, to facilitate carry out imaging agents raw material supplement next time, avoids the intersection of imaging agents raw material Infection.In other embodiments, the number of imaging agents stock bottle 71 can also be the quantity such as six or eight, according to reality Need specifically to adjust.

The setting identical with the structure of the first multiple-way valve 20 of 4th multiple-way valve 72.Unlike, the 4th multiple-way valve 72 realizes the Four pumps 74 are connected with multiple imaging agents stock bottles 71 successively, and the 4th multiple-way valve 72 selectes one of imaging agents stock bottle 71 connections, and duration is connected by controlling, so as to control the 4th pump 74 to take out imaging agents raw material in imaging agents stock bottle 71 The regulation of taken amount.So as to realize that the imaging agents raw material of multiple imaging agents stock bottles 71 can be configured in proportion, with symbol Close sequencing demand.

The setting identical with the structure of the first triple valve 30 of 3rd triple valve 73.3rd triple valve 73 can realize the He of the 4th pump 74 4th multiple-way valve 72 is connected, and the imaging agents raw material in multiple imaging agents stock bottles 71 can be extracted so as to the 4th pump 74, is matched somebody with somebody It is set to imaging agents.3rd triple valve 73 can realize that the 4th pump 74 connects with the 3rd reagent bottle 13, so as to which the 4th pump 74 can be with The imaging agents being configured are injected into imaging agents bottle 13.

4th pump 74 can be provided to multiple imaging agents stock bottles 71 through the 3rd triple valve 73 and the 4th multiple-way valve 72 and born Pressure, to extract the imaging agents raw material in multiple imaging agents stock bottles 71.4th pump 74 can also through the 3rd triple valve 73 to Malleation is provided in 3rd reagent bottle 13, to inject in imaging agents to the 3rd reagent bottle 13.

Further, imaging agents configuration component 70 also includes the second blender 75, and the second blender 75 is connected to the 3rd Between the reagent bottle 13 of triple valve 73 and the 3rd, the second blender 75 includes multiple second serpentine tubes 751, multiple second serpentine pipes Road 751 joins end to end, and is communicated between the 3rd triple valve 73 and the 3rd reagent bottle 13.

The setting identical with the structure of the first blender 65 of second blender 75, what the second blender 75 injected from the 4th pump 74 Imaging agents enter row buffering by multiple second serpentine tubes 751, and increase the flowing stroke of imaging agents, so that into As a variety of imaging agents raw materials in reagent are sufficiently mixed, imaging agents reaction efficiency is lifted.

Further, in some embodiments, drive component 50 also include the 4th triple valve 53, the 5th triple valve 54, Second waste liquid bottle 55 and the 3rd waste liquid bottle 56.The pipeline of 4th triple valve 53 is connected between the first pump 51 and first module 41, together When go back pipeline connect the second waste liquid bottle 55.The pipeline of 5th triple valve 54 is connected between the second pump 52 and second unit 42, simultaneously Also pipeline connects the 3rd waste liquid bottle 56.

First pump 51 connects the waste liquid bottle 55 of first module 41 or second through the 4th triple valve 53, so as to which the first pump 51 extracts the After the waste liquid that sequencing reaction has been completed in one unit 41, waste liquid can be injected to the second waste liquid bottle 55, so that the One pump 51 carries out providing negative pressure to first module 41 next time, to carry out sequencing reaction.5th triple valve 54 and the 4th 3 The identical setting of the structure of port valve 53, will not be repeated here, the identical setting with the structure of the second waste liquid bottle 55 of the 3rd waste liquid bottle 56, herein not Repeat again.

In the utility model embodiment, imaging agents configuration component 70 realizes the on-line mixing of fluid means 500 Function.It is appreciated that in some embodiments, fluid means can also not have on-line mixing function, correspondingly, will can be imaged Reagent configuration component 70 omits.It can so make that the pipeline of fluid means is simpler, Sequence Detection System more compact.

In some embodiments, fluid means 500 includes the first control unit, the first control unit electrical connection valve body group Part 10 and drive component 50 are run with application valve body component 10 and drive component 50.In this way, valve component 10 and driving can be realized The Automated condtrol of component 50, and then improve efficiency.

Specifically, in the utility model example, the first control unit electrically connects the first multiple-way valve 20, the first triple valve 30 With drive component 50, run with the first multiple-way valve 20 of control, the first triple valve 30 and drive component 50.First control unit can be with It is to include the devices such as single-chip microcomputer, calculator processor or central control processor, it is logical using the first control unit control more than first Valve 20, the first triple valve 30 and drive component 50 are run, and realize the automatic running of fluid means 500, improve efficiency.Further, In the utility model example, the first control unit also electrically connects the second multiple-way valve 62, the second triple valve 63, the 3rd multiple-way valve 66th, the 4th multiple-way valve 72, the 3rd triple valve 73, the 3rd pump 64 and the 4th pump 74 so that the operational efficiency of fluid means 500 is lifted.

In some embodiments, the method being controlled to sequencing reaction, in addition to：Determine the use of imaging device Sample in 102 pairs of first modules 41 and/or second unit 42 carries out multiple setting positions during IMAQ.It so, it is possible Shorten the image acquisition time of imaging device 102, improve efficiency.

Specifically, can input to enter the sample 300 of first module 41 and the passage of second unit 42 in imaging device 102 Initial position during row IMAQ, such as initial XY positions, and set and scheme needed for the distance moved every time and each passage As the number of collection, sequencing reaction is carried out from initial position.

Usually, each unit of reaction unit 200 includes plurality of passages to accelerate the sequencing to sample 300.Often The view data of sample 300 on individual passage is made up of multiple visuals field (Field of View, FOV).In one example, it is intended to pair Sample 300 on multiple passages of unit carries out IMAQ, and 300 FOV will be obtained by setting each passage, according to setting FOV numbers are controlled to the shift position of reaction unit 200.

In some embodiments, Figure 16 please be join, imaging device 102 includes the second control unit 502, image acquisition units 506 and secondary light source 508, the second control unit 502 sends initialization directive and driving instruction, microscope carrier 103 refer to according to initialization Order determines multiple setting positions, and figure is being carried out to the sample 300 in first module 41 and second unit 42 using imaging device 102 During as collection, microscope carrier 103 is according to multiple setting positions and driving instruction mobile response device, when the mobile response device of microscope carrier 103 200 when arriving setting position, and the second control unit 502 control secondary light source 508 emits light into first module 41 and/or second unit 42 so that sample 300 inspires detection light, and the control acquisition testing light of image acquisition units 506 to form view data.In this way, Realize and automatically controlling for IMAQ is carried out to the sample 300 in first module 41 and second unit 42.

Specifically, in some embodiments, the second control unit 502 includes host computer 510 and slave computer 512, upper Machine 510 is used to send initialization directive.Slave computer 512 is used to send driving instruction according to initialization directive.When microscope carrier 103 moves When reaction unit 200 arrives setting position, secondary light source 508 is controlled to emit light into sample 300 so that sample using slave computer 512 300 inspire detection light, and the control acquisition testing light of image acquisition units 506 to form view data.Image acquisition units 506 are used to view data being directly transferred to host computer 510.In this way, the data that can reduce host computer 510 and slave computer 512 pass Defeated number, while view data is directly transferred to host computer 510, realizes quick sequencing.

In some embodiments, microscope carrier 103 directly carries reaction unit 200, and microscope carrier 103 controls reaction unit 200 Movement in Sequence Detection System, microscope carrier 103 include position calculation unit, and each mobile response is calculated according to initialization directive The setting position of device 200 is with sequencing process mobile response device.Such as in high through-put sequence measure, one time sequence is surveyed Surely the view data of sample 300 of multiple setting positions need to be gathered, according to initialization directive, it is anti-then to calculate driving every time for microscope carrier 103 The setting position of device 200 is answered, when receiving driving instruction, reaction unit 200 is moved to according to each setting position Image acquisition units 506 can gather the region of image.It is preferred that microscope carrier 103 can realize the motion of the direction of principal axis of XYZ tri- with mobile anti- Device 200 is answered to setting position.

In other embodiment, reaction unit 200 can be placed in another supporting table, and microscope carrier 103 is by driving branch Support platform carrys out mobile response device 200 to setting position.

In some embodiments, image acquisition units 506 include camera 108 to convert optical signal into electric signal. In one example, image acquisition units 506 include light path module and camera 108, and reaction unit 200 is positioned on microscope carrier 103, position In the thing side of light path module, camera 108 is located at the image side of light path module.Light path module can be microscope, and microscope may include State the object lens 110 of the camera lens module 104 of embodiment.

In some embodiments, image acquisition units 506 are used to receive initialization directive and be opened according to initialization directive Open.In this way, image acquisition units 506 are in opening upon initialization, make the acquisition testing light of image acquisition units 506 Speed faster.

In some embodiments, host computer 510 is sent initialization directive to image by method wirelessly or non-wirelessly Collecting unit 506 and the view data for receiving the transmission of image acquisition units 506.In this way, realize host computer 510 and IMAQ list Data transfer between member 506.

Specifically, the data transfer mode between host computer 510 and image acquisition units 506 can be that WLAN passes Defeated or Bluetooth transmission, can also be universal serial bus transmission.Certainly, in other embodiments, it is not limited to above-mentioned Transmission means, suitable transmission means can be selected according to the actual requirements.

In some embodiments, slave computer 512 includes input/output end port, and input/output end port is used to export first Transistor-transistor logic level signal (ttl signal) with control secondary light source 508 launch light, and control image acquisition units 506 acquisition testing light.

In this way, slave computer 512 controls secondary light source 508 and image by the first transistor-transistor logic level signal Collecting unit 506, reduce the call duration time between slave computer 512 and secondary light source 508 and image acquisition units 506, enter one Step is quick to obtain image, realizes quick sequencing.

Specifically, in one example, secondary light source 508 sends the laser of specific wavelength, irradiates first module 41 and the Sample 300 on two units 42, the fluorescence group in sample 300 is set to send fluorescence, the fluorescence is as detection light, IMAQ list Member 506 gathers the fluorescence to form view data.

Further, transistor-transistor logic level signal transmission rate is Microsecond grade, is compared in correlation technique Communicated by serial ports, transistor-transistor logic level signal makes slave computer 512 and secondary light source 508 and IMAQ High-speed traffic is realized between unit 506, reduces the call duration time between slave computer 512 and all parts, is advantageously implemented fast The sequencing of speed, and the imaging device 102 of the utility model embodiment completes a wheel sequencing, can complete one The collection of setting position image, the shortening of accumulative call duration time is more notable after being repeated several times.

In some embodiments, in 506 acquisition testing light of image acquisition units, when reaching image acquisition units 506 The setting time for exposure after, the second control unit 502 control secondary light source 508 close.In this way, the second control unit 502 controls Secondary light source 508 lights within the time for exposure of image acquisition units 506, is closed after end exposure so that IMAQ list The image of the collection of member 506 is apparent, and has saved the energy.

Specifically, in some embodiments, slave computer 512 controls secondary light source 508 to close.

Further, in some embodiments, the time for exposure can be set in several ways, such as is according to circumstances carried out It is artificial to set, analogue exposure process is either carried out before sequencing and obtains the optimum time for exposure or passes through algorithm meter Calculate proper exposure time value.Certainly, in other embodiments, the above method setting time for exposure is not limited to, can be in reality In the appropriate method setting time for exposure is selected according to actual conditions.

In some embodiments, slave computer 512 includes input/output end port, and input/output end port is used to export second Transistor-transistor logic level signal is to control secondary light source 508 to close.In this way, slave computer 512 passes through input/output terminal Mouth output second transistor-transistor logic level signal closes secondary light source 508, reduces slave computer 512 and secondary light source Call duration time between 508, it is advantageously implemented quick sequencing.

In some embodiments, after the closing of secondary light source 508, the second control unit 502 control microscope carrier 103 will react Device 200 is moved to next setting position to complete the collection of setting position view data.In this way, imaging device 102 is to moving Each setting position of dynamic reaction unit 200 is gathered image one by one, realizes high-flux sequence.

Specifically, in some embodiments, after the closing of secondary light source 508, slave computer 512 sends driving instruction again To microscope carrier 103.Further, when completing the collection of view data corresponding to all setting positions, slave computer 512 is used to send END instruction is to host computer 510 to complete the IMAQ of reaction unit 200 unit.

In some embodiments, image acquisition units 506 are connected with host computer 510, and image acquisition units 506 often gather To the view data of a setting position, view data is transmitted to host computer 510, slave computer 512 and closed in secondary light source 508 Afterwards, sending driving instruction to microscope carrier 103 makes microscope carrier 103 that reaction unit 200 is moved into next setting position, slave computer 512 It is transmitted without waiting for view data further to shorten the sequencing time.

In some embodiments, driving instruction is pulse signal.In this way, the second control unit 502 passes through pulse signal Form send driving instruction to microscope carrier 103, reduce the call duration time between the second control unit 502 and microscope carrier 103, favorably In realize rapid serial determine.

In some embodiments, Figure 16 please be join, image acquisition units 506 include chasing after burnt module 516 and object lens 110, chased after Burnt module 516 controls object lens 110 and/or reaction unit 200 to be moved along the optical axis of object lens 110 according to initialization directive, to determine to scheme Optimal focusing position during IMAQ is carried out to sample as collecting unit 506, when carrying out IMAQ, chases after burnt module 516 Keep the distance of object lens 110 and sample corresponding to optimal focusing position constant.In this way, when each on sample needs to gather image Setting position not when on same X/Y plane, adjusted by chasing after burnt module 516 between object lens 110 and reaction unit 200 Distance, image acquisition units 506 are made to collect the picture rich in detail of sample on different X/Y plane setting positions.

Specifically, in some embodiments, the distance of object lens 110 and sample is object distance.Host computer 510 refers to initialization Order is sent to burnt module 516 is chased after, and is made to chase after burnt module 516 to start and is chased after burnt function automatically.In one example, moved along objective lens optical axis It can be regarded as moving along Z axis.

Burnt module 516 is chased after according to initialization directive, can control object lens 110 relative to the movement of reaction unit 200 so that phase Machine 108 can be into clearly sample image.It is determined that camera 108 is into burnt module 516 after clearly sample image, is chased after, to perform lock burnt Function, that is to say, that when the sample position of required collection changes and cause the distance of object lens 110 and sample to change When, chase after burnt module 516 and compensate variable quantity by controlling the movement of object lens 110 so that sample image formed by camera 108 is all the time Keep clear.

Alleged optimal focusing position corresponds to a pre-determined distance between object lens and sample, and alleged pre-determined distance can be One fixed numbers or fixed range, the correlation of attributes with imaging.In one example, the quality parameter to photograph image is passed through Make to limit in advance, optimal focusing position can be determined by ramping constraint, so that in the optimal focusing position IMAQ institute The quality of the image obtained reaches parameter set in advance.

It please join Figure 17, the utility model embodiment provides a kind of Systems for optical inspection 600, and Systems for optical inspection 600 wraps Control device 601, imaging device 102 and bogey 100 are included, bogey 100 includes temperature control device 301 and microscope carrier 103, into Picture device 102 includes camera lens module 104, and camera lens module 104 includes optical axis OP, and microscope carrier 103 is used to carry sample 300, optics inspection Examining system 600 is preset with the moving range of camera lens module 104, and moving range is determined by the first setting position and the second setting position Go out, the setting of the second setting position is relevant with the position of sample 300 and the depth of field, and temperature control device 301 is used to adjust sample 300 Temperature, control device 601 are used for：

Before IMAQ is carried out to sample 300 using imaging device 102 or imaging device 102 is being utilized to sample During 300 progress IMAQ, the scope for the temperature fluctuation for allowing sample 300 is set using temperature control device 301, so that camera lens module 104 are located in preset range along optical axis OP positional fluctuation scope.

It should be noted that in any of the above-described embodiment and embodiment to the technical characteristic of imaging method and beneficial to effect The explanation and illustration of fruit is also applied for the Systems for optical inspection 600 of present embodiment, to avoid redundancy, no longer exhibition in detail herein Open.

It please join Figure 18, the utility model embodiment provides a kind of Sequence Detection System 700, and sequencing is reacted and carried out Control, Sequence Detection System 700 include Systems for optical inspection 600, and Systems for optical inspection 600 includes control device 601, imaging dress 102 and bogey 100 are put, imaging device 102 includes camera lens module 104, and camera lens module 104 includes optical axis OP, bogey 100 include temperature control device 301 and microscope carrier 103, and microscope carrier 103 is used to carry sample 300, and Systems for optical inspection 600 is preset with camera lens The moving range of module 104, moving range are determined that the second setting position is set by the first setting position and the second setting position Fixed relevant with the position of sample 300 and the depth of field, temperature control device 301 is used for the temperature for adjusting sample 300, and control device 601 is used In carrying out IMAQ to sample 300 using imaging device 102, and it is used for：

Enter before sequencing reaction is carried out using Sequence Detection System 700 or using Sequence Detection System 700 During row sequencing, the scope for the temperature fluctuation for allowing sample 300 is set using temperature control device 301, so that the edge of camera lens module 104 Optical axis OP positional fluctuation scope is located in preset range.

It should be noted that the skill of the method that reaction is determined to control sequence in any of the above-described embodiment and embodiment The explanation and illustration of art feature and beneficial effect is also applied for the Sequence Detection System 700 of present embodiment, to avoid redundancy, This is no longer developed in details.

In some embodiments, Systems for optical inspection 600 is preset with the scope and camera lens mould of the temperature fluctuation of sample 300 Corresponding relation of the group 104 along optical axis OP positional fluctuation scope, control device 601 are used to, according to corresponding relation, control camera lens mould Group 104 is located in preset range along optical axis OP positional fluctuation scope.

In some embodiments, the corresponding relation includes：

When setting the scope of the temperature fluctuation for allowing the sample as ± 10 DEG C, position of the camera lens module along the optical axis Put fluctuation range ± 8 micron；

When setting the scope of the temperature fluctuation for allowing the sample as ± 5 DEG C, position of the camera lens module along the optical axis Fluctuation range is put as ± 4 microns；

When setting the scope of the temperature fluctuation for allowing the sample as ± 1.5 DEG C, the camera lens module is along the optical axis Positional fluctuation scope is ± 1 micron；

When setting the scope of the temperature fluctuation for allowing the sample as ± 0.5 DEG C, the camera lens module is along the optical axis Positional fluctuation scope is ± 0.5 micron.

In some embodiments, imaging device 102 includes focusing module 106, using imaging device 102 to sample 300 IMAQ is carried out, control device 601 is used for：Using focusing module 106 and camera lens module 104 is focused to sample 300.

In some embodiments, IMAQ is carried out to sample 300 using imaging device 102, control device 601 is used In：Sample 300 is carried out using camera lens module 104 to chase after Jiao.

In some embodiments, control device 601 is used for：

Using focusing, module 106 is emitted light on the sample 300 being placed on microscope carrier 103；

Camera lens module 104 is set to be moved to the first setting position along optical axis OP；

Camera lens module 104 is set to move and judge along optical axis OP to sample 300 with the first setting step-length from the first setting position Whether focusing module 106 receives the light reflected by sample 300；

When module 106 of focusing receives the light reflected by sample 300, make camera lens module 104 to be walked less than the first setting The second long setting step-length moves along optical axis OP and carries out IMAQ to sample 300 using imaging device 102, and judges to be imaged Whether the sharpness value for the image that device 102 is collected reaches given threshold；

When the sharpness value of image reaches given threshold, the current location of camera lens module 104 is preserved as save location.

In some embodiments, focusing module 106 includes the first light source 116 and optical sensor 118, the first light source 116 For emitting light into sample 300, optical sensor 118 is used to receive the light reflected by sample 300.

In some embodiments, when module 106 of focusing receives the light reflected by sample 300, control device 601 is used In：

Camera lens module 104 is set to set step-length along optical axis with the 3rd less than the first setting step-length and more than the second setting step-length OP moves to sample 300, and the light intensity of the light received according to focusing module 106 calculates the first light intensity parameter, judges first Whether light intensity parameter is more than the first setting Intensity threshold；

When the first light intensity parameter is more than the first setting Intensity threshold, enter to exercise camera lens module 104 with the second setting step-length Moved along optical axis OP and IMAQ is carried out to sample 300 using imaging device 102, and judge that imaging device 102 is collected The sharpness value of image whether reach given threshold.

In some embodiments, focusing module 106 includes two optical sensors 118, and two optical sensors 118 are used to connect The light reflected by sample 300 is received, the first light intensity parameter is the average value of the light intensity for the light that two optical sensors 118 receive.

In some embodiments, when module 106 of focusing receives the light reflected by sample 300, control device 601 is used In：

Camera lens module 104 is set to set step-length along optical axis with the 3rd less than the first setting step-length and more than the second setting step-length OP moves to sample 300, and the light intensity of the light received according to focusing module 106 calculates the first light intensity parameter, judges first Whether light intensity parameter is more than the first setting Intensity threshold；

When the first light intensity parameter is more than the first setting Intensity threshold, make camera lens module 104 with less than the 3rd setting step-length And moved more than the 4th setting step-length of the second setting step-length along optical axis OP to sample 300, and received according to focusing module 106 The light intensity of light calculate the second light intensity parameter, judge whether the second light intensity parameter is less than the second setting Intensity threshold；

When the second light intensity parameter is less than the second setting Intensity threshold, enter to exercise camera lens module 104 with the second setting step-length Moved along optical axis OP and IMAQ is carried out to sample 300 using imaging device 102, and judge that imaging device 102 is collected The sharpness value of image whether reach given threshold.

In some embodiments, focusing module 106 includes two optical sensors 118, and two optical sensors 118 are used to connect The light that receipts are reflected by sample 300, the average value of the light intensity for the light that the first light intensity parameter receives for two optical sensors 118, two The light intensity for the light that individual optical sensor 118 receives has the first difference, and the second light intensity parameter is the first difference and setting compensation value Difference.

In some embodiments, control device 601 is used for：When making camera lens module 104 with the second setting step-length movement, Judge whether the first sharpness value of the pattern corresponding to the current location of camera lens module 104 is more than the previous position of camera lens module 104 Put the second sharpness value of corresponding image；

It is more than the acutance difference between the second sharpness value and the first sharpness value and the second sharpness value in the first sharpness value to be more than When setting difference, camera lens module 104 is set to continue to move along optical axis OP to sample 300 with the second setting step-length；

It is more than the acutance difference between the second sharpness value and the first sharpness value and the second sharpness value in the first sharpness value to be less than When setting difference, camera lens module 104 is set to continue with the 5th setting step-length less than the second setting step-length along optical axis OP to sample 300 Movement is so that the sharpness value for the image that imaging device 102 is collected reaches given threshold；

It is more than the acutance difference between the first sharpness value and the second sharpness value and the first sharpness value in the second sharpness value to be more than When setting difference, camera lens module 104 is set to be moved with the second setting step-length along optical axis OP away from sample 300；

It is more than the acutance difference between the first sharpness value and the second sharpness value and the first sharpness value in the second sharpness value to be less than When setting difference, camera lens module 104 is set to be moved with the 5th setting step-length along optical axis OP away from sample 300 so that imaging device 102 The sharpness value of the image collected reaches given threshold.

In some embodiments, control device 601 is used for：When camera lens module 104 moves, camera lens module 104 is judged Current location whether exceed the second setting position；

When exceeding the second setting position in the current location of camera lens module 104, stop moving lens module 104 or stop Focused.

The moving range of camera lens module 104 can be preset and preserved by Systems for optical inspection 100.In some embodiments, Control device 601 has man-machine interface, and user can set the moving range of camera lens module 104, control device by man-machine interface 601 are stored in moving range itself memory or external memory storage, in this way, making Systems for optical inspection 100 be preset with camera lens The moving range of module 104, the moving range of camera lens module 104 is read when subsequently being used for Systems for optical inspection 100.

In some embodiments, bogey 100 includes：Bottom plate 101；Microscope carrier 103 is fixed on bottom plate, microscope carrier 103 Provided with storage tank 242, storage tank 242 is equipped with sample 300, and the bottom of storage tank 242 is provided with through hole 2422, temperature control device 301 It is connected by through hole 2422 with the sample 300 in storage tank 242；Resilient support assemblies 40, temperature control device 301 are propped up by elasticity Support component 40 is resiliently supported on bottom plate 101.

In some embodiments, resilient support assemblies 40 include guiding tube 43 and elastic component 44, and temperature control device 301 includes Temperature control portion 312 and guide posts 341, guide posts 341 are arranged far from the side in temperature control portion 312 of storage tank 242, guiding tube 43 It is fixed on bottom plate 101, guide posts 341 wear elastic component 44 and guiding tube 43, and the elasticity of elastic component 44 is contradicted in the He of temperature control portion 312 Between guiding tube 43；Guiding tube 43 is linear bearing, guide posts 341 and the ball sliding contact of linear bearing.

In some embodiments, temperature control device 301 includes fixed plate 321, temperature conduction plate 322, the and of temperature control element 323 Guide posts 341, temperature control element 323 are folded between fixed plate 321 and temperature conduction plate 322, and temperature control element 323 passes with temperature Guide plate 322 and fixed plate 321 contact, and temperature conduction plate 322 is used to contact with the sample 300 being loaded in storage tank 242, guides Post 341 is arranged far from the surface of fixed plate 321 of temperature control element 323, and guide posts 341 wear resilient support assemblies 40.

In some embodiments, temperature control device 301 also includes the fixed plate 321 for being arranged far from temperature control element 323 Water-bath room 324 on surface, water-bath room 324 are arranged at intervals with guide posts 341.

In some embodiments, sequencing reaction includes the first biochemical reaction and the second biochemical reaction, and first is biochemical Reaction and the second biochemical reaction are carried out on reaction unit 200, and Sequence Detection System 700 includes fluid means 500, fluid means 500 coupled reaction devices 200, reaction unit 200 include first module 41 and second unit 42, and sample 300 is placed in first module 41 and second unit 42 on, one kind for including of defined nucleotide sequence measure reaction repeats unit as the life of the second biochemical reaction-the first Change reaction-IMAQ, control device 601 includes initial module and processing module, initial module connection processing module, utilized After initial module completes following a, b, c, processing module is used to work as to make first module 41 and second using fluid means 500 It is right using imaging device 102 while the second biochemical reaction and the first biochemical reaction of a progress sample 300 in unit 42 The sample 300 of another unit carries out IMAQ,

Initial module is used for：

A makes the sample 300 on one in first module 41 and second unit 42 carry out the first life using fluid means 500 Change reaction,

B carries out IMAQ using imaging device 102 to the sample 300 on the unit after carrying out the first biochemical reaction,

C made using fluid means 500 another in first module 41 and second unit 42 on sample 300 carry out first Biochemical reaction.

In some embodiments, a and c is carried out simultaneously, or b and c are carried out simultaneously, or b is carried out before c, or b c it After carry out.

In some embodiments, fluid means 500 includes valve component 10 and drive component 50, and drive component 50 passes through Reaction unit 200 connects valve component 10, and the sample on first module 41 and/or second unit 42 is made using fluid means 500 When product 300 carry out the first biochemical reaction and/or the second biochemical reaction, valve component 10, which is used to switch, connects different reagents, drives Dynamic component 50 makes valve component 10 export reagent to first module 41 and/or second unit 42.

In some embodiments, valve component 10 includes the first multiple-way valve 20 and the first triple valve 30, the first multiple-way valve 20 switchings connect different reagent to the first triple valve 30, the first triple valve 30 by the reagent that the first multiple-way valve 20 exports export to First module 41 and/or second unit 42.

In some embodiments, drive component 50 includes the first pump 51 and the second pump 52, and the first pump 51 is single by first Member 41 connects valve component 10, and the second pump 52 connects valve component 10 by second unit 42, makes the using fluid means 500 When sample 300 on one unit 41 and/or second unit 42 carries out the first biochemical reaction and/or the second biochemical reaction, the is utilized One pump 51 makes valve component 10 export reagent makes valve component 10 export reagent to first module 41, and/or using the second pump 52 To second unit 42.

It please join Figure 17 and Figure 18, a kind of control device 601 being controlled to imaging, for Systems for optical inspection 600, light Learning detecting system 600 includes imaging device 102 and bogey 100, and bogey 100 includes temperature control device 301 and microscope carrier 103, imaging device 102 includes camera lens module 104, and camera lens module 104 includes optical axis OP, and microscope carrier 103 is used to carry sample 300, Temperature control device 301 is used for the temperature for adjusting sample 300, and control device 601 includes：

Storage device 602, for data storage, data include computer executable program；

Processor 604, for performing computer executable program, performing computer executable program includes completing above-mentioned The method of one embodiment.

In some embodiments, processor 604 includes initial module and processing module.

A kind of computer-readable recording medium of the utility model embodiment, for storing the journey performed for computer Sequence, configuration processor include the method for completing any of the above-described embodiment.Computer-readable recording medium can include：Read-only storage Device, random access memory, disk or CD etc..

In the description of this specification, reference term " embodiment ", " some embodiments ", " schematically implementation The description of mode ", " example ", " specific example " or " some examples " etc. means with reference to the embodiment or example description Specific features, structure, material or feature are contained at least one embodiment or example of the present utility model.In this explanation In book, identical embodiment or example are not necessarily referring to the schematic representation of above-mentioned term.Moreover, the specific spy of description Sign, structure, material or feature can combine in an appropriate manner in any one or more embodiments or example.

The logic and/or step for representing or otherwise describing herein in flow charts, for example, being considered use In the order list for the executable instruction for realizing logic function, may be embodied in any computer-readable recording medium, For instruction execution system, device or equipment (system of such as computer based system including processor or it is other can be from finger The system for making execution system, device or equipment instruction fetch and execute instruction) use, or combine these instruction execution systems, device Or equipment and use.For the purpose of this specification, " computer-readable recording medium " can be it is any can include, store, communicating, Propagate or transmission program for instruction execution system, device or equipment or with reference to these instruction execution systems, device or equipment and The device used.The more specifically example (non-exhaustive list) of computer-readable recording medium includes following：With one or The electrical connection section (electronic installation) of multiple wirings, portable computer diskette box (magnetic device), random access memory (RAM), only Read memory (ROM), erasable edit read-only storage (EPROM or flash memory), fiber device, and portable light Disk read-only storage (CDROM).In addition, computer-readable recording medium, which can even is that, to print the paper of described program thereon Or other suitable media, because can then enter edlin, interpretation for example by carrying out optical scanner to paper or other media Or handled if necessary with other suitable methods electronically to obtain described program, it is then stored in computer and deposits In reservoir.

In addition, each functional unit in each embodiment of the utility model can be integrated in a processing module, Can also be that unit is individually physically present, can also two or more units be integrated in a module.Above-mentioned collection Into module can both have been realized in the form of hardware, can also be realized in the form of software function module.It is described integrated If module is realized in the form of software function module and as independent production marketing or in use, can also be stored in one In computer read/write memory medium.

Although embodiment of the present utility model has been shown and described above, it is to be understood that above-mentioned embodiment party Formula is exemplary, it is impossible to is interpreted as to limitation of the present utility model, one of ordinary skill in the art is of the present utility model In the range of above-mentioned embodiment can be changed, change, replace and modification.

Claims (16)

1. a kind of Systems for optical inspection, it is characterised in that the Systems for optical inspection includes control device, imaging device and carrying Device, the bogey include temperature control device and microscope carrier, and the imaging device includes camera lens module, and the camera lens module includes Optical axis, the microscope carrier are used to carry sample, and the Systems for optical inspection is preset with the moving range of the camera lens module, the shifting Move scope to be determined by the first setting position and the second setting position, the setting of second setting position and the position of the sample Put and the depth of field is relevant, the temperature control device is used for the temperature for adjusting the sample, and the control device is used for：

Before IMAQ is carried out to the sample using the imaging device or the imaging device is being utilized to described When sample carries out IMAQ, the scope of the temperature fluctuation of the sample is allowed using temperature control device setting, so that described Positional fluctuation scope of the camera lens module along the optical axis is located in preset range.

2. a kind of Sequence Detection System, sequencing reaction is controlled, it is characterised in that the Sequence Detection System includes Systems for optical inspection, the Systems for optical inspection include control device, imaging device and bogey, and the imaging device includes Camera lens module, the camera lens module include optical axis, and the bogey includes temperature control device and microscope carrier, and the microscope carrier is used to carry Sample, the Systems for optical inspection are preset with the moving range of the camera lens module, and the moving range is by the first setting position Determined with the second setting position, the setting of second setting position is relevant with the position of the sample and the depth of field, described Temperature control device is used for the temperature for adjusting the sample, and the control device is used to carry out the sample using the imaging device IMAQ, and be used for：

Before the sequencing reaction is carried out using the Sequence Detection System or utilizing the Sequence Detection System When carrying out the sequencing, the scope of the temperature fluctuation of the sample is allowed using temperature control device setting, so that described Positional fluctuation scope of the camera lens module along the optical axis is located in preset range.

3. system as claimed in claim 1 or 2, it is characterised in that the Systems for optical inspection, which is preset with, allows the sample Temperature fluctuation scope and positional fluctuation scope of the camera lens module along the optical axis corresponding relation, the control device For according to the corresponding relation, controlling positional fluctuation scope of the camera lens module along the optical axis to be located at the preset range It is interior.

4. system as claimed in claim 3, it is characterised in that the corresponding relation includes：

When setting the scope of the temperature fluctuation for allowing the sample as ± 10 DEG C, position ripple of the camera lens module along the optical axis Dynamic scope ± 8 micron；

When setting the scope of the temperature fluctuation for allowing the sample as ± 5 DEG C, position ripple of the camera lens module along the optical axis Dynamic scope is ± 4 microns；

When setting the scope of the temperature fluctuation for allowing the sample as ± 1.5 DEG C, position of the camera lens module along the optical axis Fluctuation range is ± 1 micron；

When setting the scope of the temperature fluctuation for allowing the sample as ± 0.5 DEG C, position of the camera lens module along the optical axis Fluctuation range is ± 0.5 micron.

5. system as claimed in claim 1 or 2, it is characterised in that the imaging device include focusing module, using it is described into As device is used for sample progress IMAQ, the control device：Utilize the focusing module and the camera lens module The sample is focused.

6. system as claimed in claim 5, it is characterised in that the control device is used for：

Emitted light into and be placed on the sample on the microscope carrier using the focusing module；

The camera lens module is set to be moved to first setting position along the optical axis；

The camera lens module is set to be moved simultaneously with sample described in the first setting step-length along the optical axis direction from first setting position Judge whether the focusing module receives the light reflected by the sample；

It is described focusing module receive by the sample reflect it is described smooth when, make the camera lens module with less than described first Second setting step-length of setting step-length moves along the optical axis and carries out IMAQ to the sample using the imaging device, And judge whether the sharpness value for the described image that the imaging device is collected reaches given threshold；

When the sharpness value of described image reaches the given threshold, the current location of the camera lens module is preserved as preservation position Put.

7. system as claimed in claim 6, it is characterised in that the institute reflected by the sample is received in the focusing module When stating light, the control device is used for：

Make the camera lens module with less than the described first setting step-length and more than the 3rd setting step-length of the described second setting step-length Moved along sample described in the optical axis direction, and the light intensity of the light received according to the focusing module calculates the first light intensity Parameter, judges whether the first light intensity parameter is more than the first setting Intensity threshold；

When the first light intensity parameter is more than the described first setting Intensity threshold, enter to exercise the camera lens module with described second Setting step-length moves along the optical axis and carries out IMAQ to the sample using the imaging device, and judges the imaging Whether the sharpness value for the described image that device is collected reaches given threshold.

8. system as claimed in claim 7, it is characterised in that the focusing module includes two optical sensors, described two Optical sensor is used to receive the light reflected by the sample, and the first light intensity parameter receives for described two optical sensors The average value of the light intensity of the light arrived.

9. system as claimed in claim 6, it is characterised in that the institute reflected by the sample is received in the focusing module When stating light, the control device is used for：

Make the camera lens module with less than the described first setting step-length and more than the 3rd setting step-length of the described second setting step-length Moved along sample described in the optical axis direction, and the light intensity of the light received according to the focusing module calculates the first light intensity Parameter, judges whether the first light intensity parameter is more than the first setting Intensity threshold；

When the first light intensity parameter is more than the described first setting Intensity threshold, make the camera lens module with less than the described 3rd Set step-length and moved more than sample described in the 4th setting step-length along the optical axis direction of the described second setting step-length, and according to institute The light intensity for stating the light that focusing module receives calculates the second light intensity parameter, judges whether the second light intensity parameter is less than Second setting Intensity threshold；

When the second light intensity parameter is less than the described second setting Intensity threshold, enter to exercise the camera lens module with described second Setting step-length moves along the optical axis and carries out IMAQ to the sample using the imaging device, and judges the imaging Whether the sharpness value for the described image that device is collected reaches given threshold.

10. system as claimed in claim 9, it is characterised in that the focusing module includes two optical sensors, described two Optical sensor is used to receive the light reflected by the sample, and the first light intensity parameter receives for described two optical sensors The average value of the light intensity of the light arrived, the light intensity for the light that described two optical sensors receive have the first difference, second light Strong parameter is the difference of first difference and setting compensation value.

11. the system as described in claim any one of 6-10, it is characterised in that the control device is used for：Make the mirror When head mould group is with the described second setting step-length movement, the of the pattern corresponding to the current location of the camera lens module is judged Whether one sharpness value is more than the second sharpness value of the described image corresponding to the prior location of the camera lens module；

It is more than in first sharpness value between second sharpness value and first sharpness value and second sharpness value When acutance difference is more than setting difference, the camera lens module is set to continue with the described second setting step-length along sample described in the optical axis direction Product move；

It is more than in first sharpness value between second sharpness value and first sharpness value and second sharpness value When acutance difference is less than the setting difference, make the camera lens module with the 5th setting step-length less than the described second setting step-length Continuation is moved along sample described in the optical axis direction so that the sharpness value for the described image that the imaging device is collected reaches institute State given threshold；

It is more than in second sharpness value between first sharpness value and second sharpness value and first sharpness value When acutance difference is more than the setting difference, make the camera lens module remote described along the optical axis with the described second setting step-length Sample moves；

It is more than in second sharpness value between first sharpness value and second sharpness value and first sharpness value When acutance difference is less than the setting difference, make the camera lens module remote described along the optical axis with the described 5th setting step-length Sample movement is so that the sharpness value for the described image that the imaging device is collected reaches the given threshold.

12. the system as described in claim any one of 6-10, it is characterised in that the control device is used for：In the camera lens When module moves, judge whether the current location of the camera lens module exceeds second setting position；

When exceeding second setting position in the current location of the camera lens module, stop the mobile camera lens module or stop Only carry out the focusing.

13. system as claimed in claim 1 or 2, it is characterised in that the bogey includes：

Bottom plate；

The microscope carrier is fixed on the bottom plate, and the microscope carrier is provided with storage tank, and the storage tank is equipped with the sample, described The bottom of storage tank is provided with through hole, and the temperature control device is connected by the through hole with the sample in the storage tank；

Resilient support assemblies, the temperature control device are resiliently supported on the bottom plate by the resilient support assemblies.

14. system as claimed in claim 13, it is characterised in that the resilient support assemblies include guiding tube and elastic component, The temperature control device includes temperature control portion and guide posts, and the guide posts are arranged far from the one of the temperature control portion of the storage tank On side, the guiding tube is fixed on the bottom plate, and the guide posts wear the elastic component and the guiding tube, the elasticity Part elasticity is contradicted between the temperature control portion and the guiding tube.

15. system as claimed in claim 13, it is characterised in that the temperature control device includes fixed plate, temperature conduction plate, temperature Control element and guide posts, the temperature control element are folded between the fixed plate and the temperature conduction plate, the temperature control element Contacted with the temperature conduction plate and the fixed plate, the temperature conduction plate is used for the institute with being loaded in the storage tank Sample contact is stated, the guide posts are arranged far from the surface of the fixed plate of the temperature control element, and the guide posts are worn If the resilient support assemblies.

16. system as claimed in claim 2, it is characterised in that sequencing reaction includes the first biochemical reaction and the Two biochemical reactions, first biochemical reaction and second biochemical reaction are carried out on reaction unit, the sequencing system System includes fluid means, and the fluid means connects the reaction unit,

The reaction unit includes first module and second unit, and the sample is placed in the first module and the second unit On, it is second the-the first biochemical reaction of biochemical reaction-image to define one kind that the sequencing reaction includes and repeat unit Collection,

The control device includes initial module and processing module, after following a, b, c is completed using the initial module, institute Processing module is stated to be used for so that work as makes one in the first module and the second unit to carry out using the fluid means While second biochemical reaction of the sample and first biochemical reaction, using the imaging device to another list The sample of member carries out IMAQ,

The initial module is used for：

A makes the sample on one in the first module and the second unit carry out first using the fluid means Biochemical reaction,

B carries out IMAQ using the imaging device to the sample on the unit after carrying out first biochemical reaction,

C made using the fluid means another in the first module and the second unit on the sample carry out the One biochemical reaction.