CN208883902U - A kind of time difference imaging culture systems - Google Patents

Abstract

The utility model discloses a kind of time differences to be imaged culture systems, belong to Embryo Culture technical field, how simple the technical problems to be solved in the utility model is, safety, individually isolation culture and label and the developmental state that can observe embryo constantly reliably are carried out to embryo, reduce number of unpacking, increase the success rate of Embryo Culture, mutually obscuring between embryo is prevented simultaneously, the technical solution of use are as follows: its structure includes incubator, cultivate box, several culture box settings are in culture upper box part, culture dish and lighting device are provided in culture box, culture dish is placed in culture box, the top of culture dish in culture box is arranged in lighting device, automatic mobile device is provided in incubator, automatic imaging device and control system, control system includes microprocessor, automatic mobile device, automatic imaging device, lighting device is equal It is connect with microprocessor.

Description

A kind of time difference imaging culture systems

Technical field

The utility model relates to Embryo Culture technical field, culture systems are imaged in specifically a kind of time difference.

Background technique

Estimate according to the World Health Organization (WHO), the couple at child-bearing age of 10-15% is perplexed by infertility in developed country.In development In country (including China), until 2002 have 1.86 hundred million women of child-bearing age to suffer from infertility.Statistical data in 2014 shows, The incidence of China's reproductive population infertility is 12.5%, and infertility number is greater than 40,000,000.Vitro fertilization-embryo implanting (in- Vitro fertilization and embryo transfer, IVF-ET) technology be current treatment infertility important hand Section.From 1978 since the first in the world " test-tube baby " is born in Britain, developed country helps pregnant birth by IVF-ET at present Baby accounted for the 1-3% of born baby sum, estimate the births of whole world test-tube baby more than 5,000,000 people.It is some The deriving technology of IVF-ET needs that all embryos of patient are individually distinguished and marked, the case where in order to each embryo into Row analysis and tracking.For example, implementing preimplantation embryo genetic diagnosis (preimplantationgenetic Diagnosis, PGD) during technology, need the embryo all to patient individually to be cultivated and marked, then from this The a small amount of embryo of biopsy carries out genetic analysis in a little embryos, so that it is determined that whether these embryos suffer from genetic disease, only Patient's body can be implanted by diagnosing normal embryo, and abnormal embryo not can be carried out transplanting.At this point, in incubation The mutually isolated and label of embryo is then very crucial, if embryo obscures during culture, it is likely that cause different Normal embryo is implanted into patient's body, and birth suffers from the offspring of hereditary disease.

With the development of assisted reproductive technology, how more accurately to pick out the embryo with potentiality of development and transplants, And obtaining good Clinical Pregnancy Rate in is always the hot topic that field of reproduction is inquired into.Currently, selecting traditional morphology mostly Appraisement system carries out the screening of embryo, observes a series of Static State Index at specific time point, form including ovum mother embryo, Maturity, early stage protokaryon number, size, symmetrical degree, brephic embryo's number, the symmetrical degree of blastomere, fragment rate etc. And the blister cavities of blastula stage by stages, interior embryo group and trophoderm embryo number etc..The embryo of current routine culture system comments Valence method is known together according to Istanbul, is observed at specific time point embryo, according to Morphologic Parameters and development Situation screens embryo.It is artificial to choose particular point in time to be observed by the dynamic process of embryonic development, exist certain Limitation；In addition it unpacks repeatedly, causes the temperature, humidity and gas content of Embryo Culture environment to change, to Embryo Culture ring Border has an impact；Meanwhile there are subjective factors by embryo appraiser, expose traditional appraisement system drawback gradually.

Human embryonic in vitro culture is carried out in culture dish, and it is female that laboratories in vitro fertilization most of at present carry out ovum The in vitro culture of embryo and embryo are using common Embryo Culture ware (such as 35mm, 60mm or 100mm Petri dish).Into The patient of row IVF-ET treatment often has multiple embryos to need to carry out in vitro culture, since the form of each embryo is closer to, Directly embryo can not be marked.If necessary to individually be distinguished for each embryo, most common method is to train The droplet for preparing respective numbers in ware according to the quantity of embryo is supported, 1 embryo is placed in each droplet, then in each droplet Upper reference numerals.Since human embryos are close to spherical shape, and there is the protection of oolemma, embryo will not occur as common embryo The same adherent fixed growth is very easy to move in culture medium.In addition, culture medium and paraffin oil are liquid, hold The drift of droplet easily occurs.In addition, there have the culture dish (such as WOW culture dish) of some commercializations that can have at ware bottom at present to be several small It hole can be with individually placed each embryo, in order to be delayed by growth course of the microcam to each embryo (timelapse) it is imaged.But these apertures are not mutually isolated between embryo under the same culture medium droplet , and aperture is shallower also smaller, and slight shaking this may result in embryo and float out from aperture.Compared to each droplet list Solely for culture, it is easier to embryo be caused to obscure.The method insured the most is to prepare a culture dish for each embryo and carry out Individually label and culture, but when patient's embryo's quantity is more (such as 10-20 is a even more), then need largely to train Ware is supported, this will dramatically increase cost, operating time and difficulty.Therefore, how simply, it is safe and reliable to embryo carry out Individually isolation culture and label, prevent from mutually obscuring the technical problem for needing to solve at present between embryo.

There is also staff's skilled operation problems for human embryonic in vitro culture, by fertilized eggs from by sperm transfer It moves on to and continues growth and development in the droplet of culture dish, until third day embryo freezes or embryo transfer.Suffer from third day After embryo is transplanted, freezes or continue culture by person's selection, continue to cultivate to the embryo of blastaea to still need to shift again Into new Embryo Culture ware because first day to nutritional ingredient required for third day and third day to embryonic development in the 6th day It is different, it is necessary to which that the culture dish more renewed can just guarantee that embryonic development is good.Therefore it needs to make a large amount of Embryo Culture daily Ware, while also needing to guarantee that embryo medium different in each Embryo Culture ware will not malfunction, most embryos training at present It supports and changes liquid manually, reduce the success rate of Embryo Culture to a certain extent, it is achieved that the changing liquid automatically of culture solution is also to need The problem to be solved.

Summary of the invention

The technical assignment of the utility model is to provide a kind of time difference imaging culture systems, to solve how simple, peace Entirely, individually isolation culture and label and the developmental state that can observe embryo constantly reliably are carried out to embryo, reduces and unpacks Number, increases the success rate of Embryo Culture, at the same prevent between embryo the problem of mutually obscuring.

The technical assignment of the utility model realizes in the following manner, and culture systems, including culture is imaged in a kind of time difference Case, culture box, several culture box settings are provided with culture dish and lighting device in culture upper box part, the culture box, train Feeding ware is placed in culture box, and the top of culture dish in culture box is arranged in lighting device, is provided in incubator and is automatically moved dress Set, automatic imaging device and control system, the control system include microprocessor, the automatic mobile device for drive from Dynamic imaging device realizes three-dimensional movement, and the automatic imaging device is used to that the sample in culture dish to be imaged, automatic mobile device, Automatic imaging device, lighting device are connect with microprocessor；

At least one culture region, and the changing liquid automatically mechanism with culture regional connectivity are provided on the culture dish.

Preferably, the control system further includes image capture analysis module, gas supply module, display and environmental monitoring Module, microprocessor are separately connected image capture analysis module, gas supply module, display and environment monitoring module；

Wherein, image capture analysis module is used to capture the image of embryo and progress in culture dish by automatic imaging device Analysis；

Module is supplied to be used to inflate to incubator；

Environment monitoring module is used for the environmental change of real-time monitoring incubator and carries out deviation early warning；

Display is for showing the analysis of ambient conditions and image capture analysis module to embryo in culture dish in incubator As a result.

Preferably, the culture region includes culture pond and waste liquid pool, changing liquid automatically mechanism includes new culture solution storage Case, peristaltic pump one and peristaltic pump two, new culture solution storage box, peristaltic pump one, culture pond, peristaltic pump two, waste liquid pool pass through pipeline It is sequentially communicated；Peristaltic pump two is mounted on the upper side wall of culture box, and the interface of peristaltic pump two, using being threadedly coupled, is trained with culture dish It is convenient to support ware replacement.

More preferably, the changing liquid automatically mechanism further includes waste liquid collecting box and peristaltic pump three, waste liquid pool, peristaltic pump three and useless Liquid collecting box is sequentially communicated by pipeline；It is provided with switch valve one on pipeline between new culture solution storage box and peristaltic pump one, It is provided with switch valve two on pipeline between culture pond and peristaltic pump two, is provided on the pipeline between waste liquid pool and peristaltic pump three Switch valve three.

More preferably, the culture region further includes the service sink one and service sink two around culture pond setting, service sink one It is used to after embryo's cleaning be placed into culture pond with service sink two.

More preferably, the culture bottom of pond portion is provided with several culturing room, and culturing room is circumferentially distributed and the cross of culturing room Section is in inverted ladder shape, and culturing room top opens up fluted, and bottom portion of groove is in arc-shaped, and groove is for placing embryo to be cultivated Tire.

Preferably, be provided with heating layer above and below the culture dish, heating layer setting in culture box and The both ends of heating layer are separately fixed on the two sides inner wall of culture box box cover.

Preferably, the lighting device includes lamp bracket, LED illumination mould group and condenser mould group, lamp bracket setting is being cultivated The box cover inner wall of box, LED illumination mould group and condenser mould group are arranged on lamp bracket and condenser mould group is located at LED illumination mould group Lower section.

More preferably, the LED illumination mould group includes pedestal, and multiple LED chips are provided on pedestal, and LED chip is in positive six The distribution of side shape.

More preferably, the condenser mould group includes connection frame, optically focused microscope group one and optically focused microscope group two, optically focused microscope group one and poly- The middle position of lamp bracket is arranged in by connection frame for light microscopic group two and optically focused microscope group one is located at two top of optically focused microscope group；Optically focused microscope group One and optically focused microscope group two include frame and condenser, condenser is in the frame and condenser extends frame, condenser The condenser of group one and the condenser of condenser two are oppositely arranged.

Preferably, the automatic mobile device includes that guide rail, device of rotation driving, mobile drive deivce and setting are being led Sliding block on rail, guide rail is arranged in incubator and the both ends of guide rail are fixed on the two sides inner wall of incubator, and rotation is driven Dynamic device is arranged in upper end of slide block, and automatic imaging device is mounted on the output end of device of rotation driving by connecting shaft；It is mobile to drive The side of sliding block is arranged in dynamic device and mobile drive deivce passes through the side wall of incubator and is fixed on the lateral wall of incubator.

More preferably, the mobile drive deivce uses cylinder, and the cylinder body end of cylinder is fixed on the lateral wall of incubator, gas The tailpiece of the piston rod of cylinder passes through the side wall of incubator and is fixedly connected with sliding block；Cylinder block is arranged by mounting plate in incubator Lateral wall on；Wherein, driving device can also use motor and rack pinion structure.

More preferably, the device of rotation driving uses servo motor, and servo motor is fixed at upper end of slide block, servo electricity Machine is rotatablely connected automatic imaging device by connecting shaft.

Preferably, the automatic imaging device includes camera mounting bracket, camera and limited remote correction object lens, camera installation Frame is rotatablely connected servo motor output end by connecting shaft, and camera is arranged in camera mounting bracket upper end, and limited remote correction object lens are set It sets in camera upper end and limited remote correction object lens is located at below culture dish.

The time difference imaging culture systems of the utility model have the advantage that compared with prior art

(1), time difference imaging technique is a kind of imaging technique that moment exposure is continuously shot, and has high resolution ratio, this Utility model is using time difference imaging technique to embryo's screening and assessment, and real time monitoring carries out dynamic observation to embryo, and using shape State kinetic parameter quantifies embryo development procedure, it is made more to objectify；

(2), automatic imaging device is combined with incubator, effectively reduces unpacking frequency, it is ensured that embryo sends out during observation The stability for educating environment, during supplementary reproduction, provide one can temperature control (close to human body temperature) environment, have to embryo Tire culture environment monitors in real time, to the function of embryonic development state captured in real-time analysis, realizes automation comprehensive detection embryo hair Educate situation；

(3), multiple culture boxes can be placed on the incubator of the utility model simultaneously, a culture is placed in each culture box Ware, at least one culture pond in each culture dish, one circle culturing room of setting in culture pond not only can be simultaneously to the embryo of different people Tire is cultivated and is not easy to obscure, and can carry out the culture of multiple embryos to the same person and be not easy to obscure, Substantially increase the efficiency and success rate of Embryo Culture；

(4), culture dish can realize changing liquid automatically by changing liquid automatically mechanism, during avoiding manual operation, to embryo It damages, influences the success rate of Embryo Culture.

Detailed description of the invention

The present invention will be further described with reference to the accompanying drawing.

Attached drawing 1 is the structural schematic diagram of incubator；

Attached drawing 2 is the schematic diagram of internal structure of incubator；

Attached drawing 3 is the structural schematic diagram for cultivating box；

Attached drawing 4 is the structural schematic diagram of lighting device；

Attached drawing 5 is the structural schematic diagram of LED illumination mould group；

Attached drawing 6 is the structural schematic diagram of condenser mould group；

Attached drawing 7 is the structural schematic diagram of culture dish and changing liquid automatically mechanism；

Attached drawing 8 is the structural schematic diagram of culturing room；

Attached drawing 9 is the structural block diagram of control system；

Attached drawing 10 is the flow chart of image capture analysis module work.

In figure: 1, incubator, 2, culture box, 3, culture dish, 4, culture oil, 5, microprocessor, 6, image capture analysis Module, 7, gas supply module, 8, display, 9, environment monitoring module, 10, new culture solution storage box, 11, culture pond, 12, waste liquid Pond, 13, waste liquid collecting box, 14, peristaltic pump one, 15, peristaltic pump two, 16, peristaltic pump three, 17, service sink one, 18, service sink two, 19, culturing room, 20, groove, 21, heating layer, 22, lamp bracket, 23, LED illumination mould group, 23-1, pedestal, 23-2, LED chip, 24, Condenser mould group, 24-1 connection frame, 24-2, optically focused microscope group one, 24-3 optically focused microscope group two, 24-4, frame, 24-5 condenser, 25, Guide rail, 26, sliding block, 27, connecting shaft, 28, cylinder, 29, servo motor, 30, camera mounting bracket, 31, camera, 32, limited remote school Positive lens, 33, liquid crystal touch screen, 34, mounting plate, 35, embryo, 36, culture solution.

Specific embodiment

A kind of imaging of time difference culture systems of the utility model are made referring to Figure of description and specific embodiment following detailed Carefully illustrate.

Embodiment 1:

As shown in Fig. 1, the utility model the time difference be imaged culture systems, structure include incubator 1 and culture box 2, Six culture boxes 2 are mounted on 1 top of incubator, cultivate in box 2 and are equipped with culture dish 3 and lighting device, culture dish 3 is placed in culture In box 2, lighting device is mounted on the top of culture dish 3 in culture box 2, be equipped in incubator 1 automatic mobile device, automatically at As device and control system, control system includes microprocessor 5, and automatic mobile device is for driving automatic imaging device to realize three Dimension movement, automatic imaging device are used to that the sample in culture dish 3 to be imaged, automatic mobile device, automatic imaging device, illumination dress It sets and is connect with microprocessor 5；Culture dish 3 is equipped with culture region, and the changing liquid automatically mechanism with culture regional connectivity.Its In, incubator 1 has occupied area small, and bearer cap is big, and design should be as compact as possible, can multiple-bearer patient populations (mesh as far as possible The time difference incubator maximum bearing capacity of preceding design can reach 16 culture boxes)；1 configuration of incubator uses and independently cultivates box 2, and every Patient's Embryo Culture is independent of each other, while should reduce influence when loading culture dish to whole culture environment as far as possible.Incubator 1 Liquid crystal touch screen 33 is installed, liquid crystal touch screen 33 can be directly on incubator 1 to patient's embryonic development situation at upper inclined surface And current condition of culture (setting values and actual value of the parameters such as temperature, CO2 concentration, oxygen concentration) is observed, Yi Jidan Point shooting photo, and photo is played out and reset.Incubator provides complete period closed environment for Embryo Culture, can carry out certainly Dynamic temperature, humidity, adjustment of gas concentration avoid the change of environment temperature, humidity and gas content to the shadow of embryo's mechanism study It rings.HEPA/VOC filter is combined by three built-in gas mixing cabins, highly stable and controllable culture ring is provided for embryo Border.The monitoring of stringent Environment features and the isolation loading area design of unique culture dish 3 ensure to open hatch door addition or move comprehensively Culture environment when except culture dish in the culturing room of inside will not change.

Embryo's amount as much as possible (mating culture dish list of the time difference incubator designed at present is reserved on culture dish 3 for patient Ware can at most carry 16 pieces of embryos), and addition patient information bar code/two dimensional code identification label position is reserved on culture dish It sets, patient information registration can be automatically performed and read.16 micropore culture dishes can be realized the uniformity of 6 micropores of culture dish, gas-tight Bubble and changing liquid automatically.

As shown in Fig. 7, culture dish 3 can carry out individually isolation training to all embryos of patient in a culture dish It supports and marks, avoid mutually obscuring between embryo, while changing liquid automatically mechanism can reduce and manually change the cumbersome of liquid.Cultivation region Domain is located at ware body inside bottom surface, cultivates remaining by outside enclosing and intermediate enclosing and is divided into four, respectively culture pond 11, service sink One 17, service sink 2 18 and waste liquid pool 12, service sink 1 and service sink 2 18 are used to be placed into culture pond after embryo's cleaning In 11, isolation height 1mm.Ware lid is additionally provided on culture dish 3, a height of 6mm of ware lid, outer diameter 41mm, with a thickness of 1mm, internal diameter is 39mm；The a height of 11mm of culture dish, outer diameter 38mm, culture dish edge enclosing thickness internal diameter is 1mm, and culture dish bottom is with a thickness of 1mm. Culture pond 11, service sink 1, service sink 2 18 and 12 outer diameter of waste liquid pool in culture dish are 9mm, internal diameter 7.5mm.Whole Changing liquid automatically mechanism is equipped in the intermediate culture pond 11 of a culture dish, changing liquid automatically can reduce embryo's exposure outside incubator 1 Time, influence the quality and activity of embryo.Changing liquid automatically mechanism includes new culture solution storage box 10, peristaltic pump 1 and wriggles 2 15 are pumped, new culture solution storage box 10, peristaltic pump 1, culture pond 11, peristaltic pump 2 15, waste liquid pool 12 are successively connected by pipeline Logical, peristaltic pump 2 15 is mounted on the upper side wall of culture box 1, and the interface of peristaltic pump 2 15 is used with culture dish 3 to be threadedly coupled, with Just culture dish 3 is replaced.The volume of waste liquid pool 12 is about three times of intermediate culture pond 11, prevent liquid outlet pipe peristaltic pump 2 15 by Pollution.The material of culture dish 3 is the plastics of various biological safety levels, preferably poly- such as polypropylene, polystyrene or polycarbonate Propylene.Changing liquid automatically mechanism further includes waste liquid collecting box 13 and peristaltic pump 3 16, waste liquid pool 12, peristaltic pump 3 16 and waste collection Case 13 is sequentially communicated by pipeline；Switch valve one, training are installed on pipeline between new culture solution storage box 10 and peristaltic pump 1 It supports and switch valve two is installed on the pipeline between pond 11 and peristaltic pump 2 15, pacify on the pipeline between waste liquid pool 12 and peristaltic pump 3 16 Equipped with switch valve three.

As shown in Fig. 8, it is equipped with around 16 small 19,16 culturing room 19, culturing room in culture pond 11 and is equipped with number, It is observed so that single Embryo Culture marks.16 culturing room 19 are circumferentially distributed and the cross section of culturing room 19 is in inverted ladder shape, 19 top of culturing room opens up fluted 20, and 20 bottom of groove is in arc-shaped, and groove 20 is for placing embryo 35 to be cultivated.Culture It is full of culture solution 36 in room 19 and places culture oil on culture solution 36 after placing embryo 35 in 19 bottom groove 20 of culturing room 4, play the role of sealing.The volume of culturing room 19 is designed as 450ul, human fertilization ovum and cleavage stage embryo diameter about 120- 150 microns, about 200-300 microns of blastaea, each culturing room can place 1 embryo.Round mouth diameter is in culturing room 500um, axis of small circle 400um.The bottom level of culturing room is convenient for the microscopical observation in workbench bottom.

As shown in Fig. 3, heating layer 21 is mounted on above and below culture dish 3, heating layer 21 is mounted on culture box 2 Interior and heating layer 21 both ends are separately fixed on the two sides inner wall of culture 2 box cover of box.

As shown in Fig. 4, lighting device includes lamp bracket 22, LED illumination mould group 23 and condenser mould group 24, and lamp bracket 22 is installed On the upper end inner wall of the box cover of culture box 2, LED illumination mould group 23 and condenser mould group 24 is installed on lamp bracket 22 and optically focused Mirror mould group 24 is located at the lower section of LED illumination mould group 23.Wherein, the illumination of LED illumination mould group 23 is when taking pictures, each image it is lasting when Between should as short as possible (reference data 0.064s at present), intensity of illumination is as low as possible, to reduce influence of the light to embryonic development (reference: 635nm red LED light source).Lighting device is able to carry out light field imaging and dark-field imaging.

As shown in Fig. 5, LED illumination mould group 23 includes pedestal 23-1, is equipped with multiple LED chip 23- on pedestal 23-1 2, LED chip 23-2 are distributed in regular hexagon.LED illumination mould group 23 uses array LED chip, wavelength 635nm；One of them LED chip is in center, other six LED chips are in the annular that radius is 20mm.When light field imaging, middle position LED chip is lighted, and other six LED chips are closed；When dark-field imaging, the LED chip in middle position is closed, and other six LED chip is lighted, by control array LED chip switch, avoid traditional dark-field imaging need mechanical switch ring baffle or The problem of person's condenser, and it is conducive to the synchronously control that illumination light switching sequence adopts figure timing with camera, dark field LED chip generates light Beam incidence angle is 26 °, meets the requirement that numerical aperture is greater than object lens NA.Condenser is optimized for LED chip array, shines Bright field 8mm, numerical aperture 0.46 can guarantee that entire orifice plate is completely covered, and when to different embryo's imaging samples, shine Mingguang City source is not necessarily to shift position, while illuminating optical numerical aperture greater than image-forming objective lens numerical aperture, can be realized dark-field imaging.

As shown in Fig. 6, condenser mould group 24 includes two 24- of connection frame 24-1, one 24-2 of optically focused microscope group and optically focused microscope group 3, one 24-2 of optically focused microscope group and two 24-3 of optically focused microscope group are mounted on middle position and the condenser of lamp bracket 22 by connection frame 24-1 One 24-2 of group is located above two 24-3 of optically focused microscope group；One 24-2 of optically focused microscope group and two 24-3 of optically focused microscope group include frame 24-4 and Condenser 24-5, condenser 24-5 are in the frame 24-4 and condenser 24-5 extends frame 24-4, one 24- of optically focused microscope group The condenser 24-5 of 2 two 24-3 of condenser 24-5 and condenser is oppositely arranged.

As shown in Fig. 2, automatic mobile device includes that guide rail 25, device of rotation driving, mobile drive deivce and setting exist Sliding block 26 on guide rail, guide rail 25 is mounted in incubator 1 and the both ends of guide rail 25 are fixedly mounted on the two sides inner wall of incubator 1 On, device of rotation driving is mounted on 26 upper end of sliding block, and automatic imaging device is mounted on device of rotation driving by connecting shaft 27 Output end；The side of sliding block 26 is arranged in mobile drive deivce, and mobile drive deivce passes through the side wall of incubator 1 and is fixed on training On the lateral wall for supporting case 1.Automatic imaging device can be realized the automatic positioning and clear observation of micron order embryo.Mobile driving dress It sets using cylinder 28, the cylinder body end of cylinder 28 is fixed on the lateral wall of incubator 1, and the tailpiece of the piston rod of cylinder 28 passes through incubator 1 side wall and it is fixedly connected with sliding block 26；28 cylinder body of cylinder is mounted on the lateral wall of incubator 1 by mounting plate 34；Its In, driving device can also use motor and rack pinion structure.Device of rotation driving uses servo motor 29, servo electricity Machine 29 is fixedly mounted on 26 upper end of sliding block, and servo motor 29 is rotatablely connected automatic imaging device by connecting shaft 27.

Automatic imaging device includes camera mounting bracket 30, camera 31 and limited remote correction object lens 32, and camera mounting bracket 30 is logical It crosses connecting shaft 27 and is rotatablely connected 29 output end of servo motor, camera 31 is mounted on 30 upper end of camera mounting bracket, limited remote correction object Mirror 32 is mounted on 31 upper end of camera and limited remote correction object lens 32 are located at 3 lower section of culture dish.Wherein, 31 resolution ratio of camera should not be low It is not less than 2.22px/ μm in 1280*1024 pixel, monochromatic 8 bits, optical resolution, 3-7 layers of focal plane, when circulation is taken pictures Between (5 minutes, 10 minutes etc.) can be freely set.The limited remote correction object lens 32 of automatic mobile device control are directed at embryo into image position Set, Embryo Culture ware 3 illuminated by LED illumination mould group 23 as lighting source, culture dish 3 illuminated by light source after under The limited remote correction object lens 32 of side are imaged on the camera at rear, and embryo's image after being imaged by camera 31 passes through Image Acquisition Card, image processing equipment obtain more perfect image after carrying out image enhanced processing, and image is presented on the display 8, embryo Image can be assessed by image capture analysis module 6 on the display 8, classification is handled.

Imaging uses the limited remote correction object lens 32 of long reach, true field 700um, numerical aperture 0.33, times magnification 17.5 times of rate, it can satisfy the requirement to embryo samples imaging resolution.500 μm of visual fields require CCD target surface to be greater than 8.7 mm, because This camera uses scientific research grade high-quantum efficiency sCMOS camera, makes the light energy required for adopting figure be well below conventional microscopy Embryonic development provides safer optical environment.Tube lens is not provided between object lens and camera, so that limited remote correction object Mirror and camera constitute a compact automatic imaging device.Automated imaging, which is loaded onto, to be mounted on servo motor, can be moved in the direction z It is dynamic to realize focusing, minimum movement step-length 0.1um；Simultaneously can be mobile in X and Y-direction by cylinder, to different embryo samples Imaging realizes and keeps sample position constant always, can reduce injury of the mechanical movement to sample to the maximum extent.Pass through servo The movement of motor makes automatic imaging device have automatic focusing function, and realizes that single focal plane imaging and multifocal surface layer sweep imaging mould Formula (most 11 layers).For single focal plane imaging mode, realize in 5 minutes to whole embryo samples imaging observations；For 11 coke charges Surface layer sweeps imaging pattern, 10 minutes achievable imaging observations to whole embryo samples.

As shown in Fig. 9, control system further includes image capture analysis module 6, gas supply module 7, display 8 and environment prison Module 9 is surveyed, microprocessor 5 is separately connected image capture analysis module 6, gas supply module 7, display 8 and environment monitoring module 9；

Wherein, the image that image capture analysis module 6 is used to capture embryo in culture dish by automatic imaging device is gone forward side by side Row analysis, improves the accuracy rate that noninvasive embryo quality assesses classification automatically；And there is following function: 1. can identify patient's bar shaped Code/two-dimensional barcode information carries out case history verification；

2. being able to record the condition of culture (temperature, CO2 concentration, O2 concentration) of embryo；

3. intellectual analysis can be carried out to the developmental state of each piece of embryo of patient and carry out judgement scoring, when can show Between axis；

4. analysis can be compared to the developmental state of more pieces of embryos of patient simultaneously；

5. intelligent guidance doctor carries out the transplanting of embryo, freezes or abandon；

6. every piece of embryonic development picture and relevant information can export, it is made into film (AVI MOV MP4).

Image capture analysis module 6 using elimination system score-system step by step, finally obtained embryo be exactly selection can be with The embryo quality of transplanting or freezing.

The time required to as shown in Fig. 10, T5_PNF is is 5 cell stages to development from fertilization female pronucleus disappearance (unit: Hour)；S2: embryo is that moment of 3 cells by 2 cell divisions, is to 3 cell divisions in the duration of 3 cell stages Time needed for that moment of 4 cells (unit: hour).

Module 7 is supplied to be used to inflate to incubator；The plenum system for supplying module includes the following two kinds: it is straight 1. to premix gas It fills；2. pure N2 and pure CO2, is mixed by built-in mixing machine；Gas detection mouth is offered on incubator, it can external CO2 detection immediately Instrument carries out culture gas concentration monitoring comparison.

Environment monitoring module 9 for real-time monitoring incubator environmental change and carry out deviation early warning (including temperature and CO2 Concentration)；

Display 8 is for showing the analysis of ambient conditions and image capture analysis module to embryo in culture dish in incubator As a result.

Specific use process is as follows:

(1), culture box 2 is arranged at the top of incubator 1, opens the box cover of culture box 2, culture dish 3 is put into training It supports in box 2；

(2), changing liquid automatically mechanism will add new culture solution in culturing room 11, and embryo to be cultivated is placed into culturing room 11 It is inside cultivated, and covers the box cover of culture box 2；

(3), the control of microprocessor 5 starting gas supply module 7, automatic mobile device, automatic imaging device, lighting device and ring Border monitoring modular 9, the environmental change of real-time monitoring incubator 1 simultaneously carry out deviation early warning；

(4), automatic mobile device drives automatic imaging device to complete three-dimensional mobile, has been tuned into the position of automatic imaging device It sets, automatic imaging device carries out imaging to the sample in culture dish, and is transmitted to image capture analysis module 6；

(5), image capture analysis module 6 captures embryo in culture dish according to the sample imaging contexts of automatic imaging device Image and analyzed；

(6), in incubator 1 ambient conditions and image capture analysis module 6 to the captured image of embryo in culture dish and Analysis result shows that staff can make a choice according to reality by display 8.

The technical personnel in the technical field can readily realize the utility model with the above specific embodiments,.But It is it should be appreciated that the utility model is not limited to above-mentioned specific embodiment.On the basis of the disclosed embodiments, described Those skilled in the art can arbitrarily combine different technical features, to realize different technical solutions.

Except for the technical features described in the specification, it all is technically known to those skilled in the art.

Claims (14)

1. culture systems, including incubator, culture box is imaged in a kind of time difference, upper box part is being cultivated in several culture box settings, It is characterized in that, culture dish and lighting device are provided in the culture box, and culture dish is placed in culture box, and lighting device setting exists The top of culture dish in box is cultivated, is provided with automatic mobile device, automatic imaging device and control system, the control in incubator System processed includes microprocessor, and the automatic mobile device is three-dimensional mobile for driving automatic imaging device to realize, described automatic Imaging device is used to that the sample in culture dish to be imaged, and automatic mobile device, automatic imaging device, lighting device are and micro process Device connection；

At least one culture region, and the changing liquid automatically mechanism with culture regional connectivity are provided on the culture dish.

2. culture systems are imaged in the time difference according to claim 1, which is characterized in that the control system further includes that image is caught Analysis module, gas supply module, display and environment monitoring module are obtained, microprocessor is separately connected image capture analysis module, supplies Gas module, display and environment monitoring module；

Wherein, image capture analysis module is used to capture the image of embryo in culture dish by automatic imaging device and divide Analysis；

Module is supplied to be used to inflate to incubator；

Environment monitoring module is used for the environmental change of real-time monitoring incubator and carries out deviation early warning；

Display is used to show that ambient conditions and image capture analysis module to be to the analysis result of embryo in culture dish in incubator.

3. culture systems are imaged in the time difference according to claim 1 or 2, which is characterized in that the culture region includes culture Pond and waste liquid pool, changing liquid automatically mechanism include new culture solution storage box, peristaltic pump one and peristaltic pump two, new culture solution storage box, Peristaltic pump one, culture pond, peristaltic pump two, waste liquid pool are sequentially communicated by pipeline.

4. culture systems are imaged in the time difference according to claim 3, which is characterized in that the changing liquid automatically mechanism further includes giving up Liquid collecting box and peristaltic pump three, waste liquid pool, peristaltic pump three and waste liquid collecting box are sequentially communicated by pipeline；New culture solution storage box It is provided with switch valve one on pipeline between peristaltic pump one, is provided with switch valve on the pipeline between culture pond and peristaltic pump two Two, switch valve three is provided on the pipeline between waste liquid pool and peristaltic pump three.

5. culture systems are imaged in the time difference according to claim 4, which is characterized in that the culture region further includes around training The service sink one and service sink two of pond setting are supported, service sink one and service sink two are used to be placed into culture pond after embryo's cleaning In.

6. culture systems are imaged in the time difference according to claim 5, which is characterized in that the culture bottom of pond portion is provided with several A culturing room, culturing room is circumferentially distributed and the cross section of culturing room is in inverted ladder shape, and culturing room top opens up fluted, groove Bottom is in arc-shaped, and groove is for placing embryo to be cultivated.

7. culture systems are imaged in the time difference according to claim 1, which is characterized in that above and below the culture dish It is provided with heating layer, heating layer is arranged in culture box and the both ends of heating layer are separately fixed at the two sides inner wall of culture box box cover On.

8. culture systems are imaged in time difference according to claim 1 or 2, which is characterized in that the lighting device include lamp bracket, The box cover inner wall of culture box is arranged in LED illumination mould group and condenser mould group, lamp bracket, and LED illumination mould group and condenser mould group are equal It is arranged on lamp bracket and condenser mould group is located at the lower section of LED illumination mould group.

9. culture systems are imaged in the time difference according to claim 8, which is characterized in that the LED illumination mould group includes pedestal, Multiple LED chips are provided on pedestal, LED chip is distributed in regular hexagon.

10. culture systems are imaged in the time difference according to claim 8, which is characterized in that the condenser mould group includes connection Frame, optically focused microscope group one and optically focused microscope group two, optically focused microscope group one and optically focused microscope group two-way cross the interposition that lamp bracket is arranged in connection frame It sets and optically focused microscope group one is located at two top of optically focused microscope group；Optically focused microscope group one and optically focused microscope group two include frame and condenser, are gathered Light microscopic is in the frame and condenser extends frame, the condenser of the optically focused microscope group one and condenser of condenser two is opposite sets It sets.

11. culture systems are imaged in time difference according to claim 1 or 2, which is characterized in that the automatic mobile device includes Guide rail, device of rotation driving, mobile drive deivce and the sliding block being arranged on guide rail, guide rail is arranged in incubator and guide rail Both ends are fixed on the two sides inner wall of incubator, and device of rotation driving is arranged in upper end of slide block, and automatic imaging device passes through Connecting shaft is mounted on the output end of device of rotation driving；The side of sliding block is arranged in mobile drive deivce and mobile drive deivce is worn It crosses the side wall of incubator and is fixed on the lateral wall of incubator.

12. culture systems are imaged in the time difference according to claim 11, which is characterized in that the mobile drive deivce uses gas The cylinder body end of cylinder, cylinder is fixed on the lateral wall of incubator, and the tailpiece of the piston rod of cylinder passes through the side wall of incubator and fixed company Connect sliding block；Cylinder block is arranged on the lateral wall of incubator by mounting plate.

13. culture systems are imaged in the time difference according to claim 11, which is characterized in that the device of rotation driving is used and watched Motor is taken, servo motor is fixed at upper end of slide block, and servo motor is rotatablely connected automatic imaging device by connecting shaft.

14. culture systems are imaged in time difference according to claim 1 or 2, which is characterized in that the automatic imaging device includes Camera mounting bracket, camera and limited remote correction object lens, camera mounting bracket are rotatablely connected servo motor output end, phase by connecting shaft Machine setting is in camera mounting bracket upper end, and limited remote correction object lens are arranged in camera upper end and limited remote correction object lens are located at culture dish Lower section.