CN202184048U - Zebra fish action inducing system - Google Patents
Zebra fish action inducing system Download PDFInfo
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- CN202184048U CN202184048U CN2011200497374U CN201120049737U CN202184048U CN 202184048 U CN202184048 U CN 202184048U CN 2011200497374 U CN2011200497374 U CN 2011200497374U CN 201120049737 U CN201120049737 U CN 201120049737U CN 202184048 U CN202184048 U CN 202184048U
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Abstract
The utility model relates to a zebra action inducing system which comprises a visual stimulation device, a camera and a data collection unit, wherein the visual stimulation device can at least induce the eye movement reaction and the optomotor response action of a zebra fish; the camera is used for shooting the actions of the zebra fish so as to obtain video data; the data collecting unit obtains the video data from the camera and inputs the video data into an analysis system; the analysis system conducts ethological analysis on the input video data so as to obtain the ethological data of the eye movement reaction and optomotor response reaction of the zebra fish. The zebra fish action inducing system can be used for analyzing the eye movement reaction and the optomotor response reaction of the adult zebra fish, and the eye movement reaction of the infantile zebra fish.
Description
Technical field
The utility model relates to a kind of bio-analysis system, especially relates to a kind of model organism zebra fish behavior and brings out and analytical system.
Background technology
Model organism zebra fish is widely used in studies such as tumour, disease of hematopoietic system, angiocardiopathy, ophthalmology disease, ephrosis disease, bone relevant disease, epilepsy, nervous system at life science.The analysis of zebra fish behaviouristics is an important research means; But present existing system of behavior can not be used for zebra fish OKR (the optokinetic response that grows up; Eye movement reaction), OMR behaviors such as (optomotor response, optomotor responses) brings out and analyzes.
The utility model content
The purpose of the utility model is to propose a kind of zebra fish behavior to bring out and analytical system, and the OKR of the zebra fish that can grow up analyzes.
The utility model proposes a kind of zebra fish behaviouristics and brings out and analytical system, comprises visual stimulus device, camera and analytical system.The visual stimulus device can bring out eye movement reaction, the optomotor response behavior of zebra fish at least.Camera is in order to take the behavior of zebra fish, to obtain video data.Analytical system is imported said video data and is carried out the behaviouristics analysis, to obtain eye movement reaction and the optomotor response behaviouristics data of zebra fish.
In an embodiment of the utility model; Said visual stimulus device is a mechanical type visual stimulus device; It comprises motor, transmission device, grating and fluorescent tube; Said motor is used to produce rotary power, and said transmission device is used for being passed to grating to the rotation of motor and rotates to drive grating, and said fluorescent tube is used to illuminate grating and produces visual pattern.
In an embodiment of the utility model, said visual stimulus device is an arc screen stimulator, and it comprises container that holds zebra fish and the arc panel type display that surrounds this container, and this arc panel type display produces image, as the visual stimulus of zebra fish.
In an embodiment of the utility model, said analytical system is based on preset one or more templates one or more area-of-interests of selected video image is carried out template matches, obtains the eye movement angle-data of zebra fish eye movement reflex action with tracking.
In an embodiment of the utility model, said analytical system is that background correction is sheared out selected area-of-interest again and isolated target through binaryzation from video image, obtains the angle-data of zebra fish optomotor response behavior with tracking.
The utility model is owing to adopt above technical scheme, make it possible to grow up zebra fish and childhood zebra fish carry out the eye movement response analysis.Further, can also carry out the optomotor response analysis to the zebra fish that grows up, and to childhood zebra fish carry out multiple spot and follow the tracks of.
Description of drawings
Fig. 1 illustrates the behavior of the utility model one embodiment and brings out and the analytical system sketch map.
Fig. 2 illustrates the visual stimulus device sketch map of the utility model one embodiment.
Fig. 3 illustrates the control circuit figure of device shown in Figure 2.
Fig. 4 illustrates the visual stimulus device sketch map of another embodiment of the utility model.
Fig. 5 illustrates the drive circuit figure of device shown in Figure 4.
Fig. 6 illustrates the behaviouristics analytical system framework of the utility model one embodiment.
Fig. 7 illustrates the adult zebra fish OKR analysis principle of the utility model one embodiment.
Fig. 8 illustrates the adult zebra fish OKR analysis process figure of the utility model one embodiment.
Fig. 9 illustrates the adult zebra fish OKR assay surface of the utility model one embodiment.
Figure 10 illustrates the adult zebra fish OMR analysis process figure of the utility model one embodiment.
Figure 11 illustrates the adult zebra fish OMR assay surface of the utility model one embodiment.
Figure 12 illustrates zebra fish OKR analysis principle childhood of the utility model one embodiment.
Figure 13 illustrates zebra fish OKR analysis process figure childhood of the utility model one embodiment.
Figure 14 illustrates zebra fish OKR assay surface childhood of the utility model one embodiment.
Embodiment
For letting above-mentioned purpose, the feature and advantage of the utility model can be more obviously understandable, elaborate below in conjunction with the embodiment of accompanying drawing to the utility model.
Fig. 1 illustrates the behavior of the utility model one embodiment and brings out and the analytical system sketch map.With reference to shown in Figure 1, zebra fish behaviouristics system 1000 comprises: visual stimulus device 100, camera 200, data acquisition unit 300 and analytical system 400.
Camera 200 is used to gather the video information of zebra fish motion.In the embodiment of the utility model, camera 200 can use CCD or the general camera of CMOS and software to make up.In order to analyze data effectively, the acquisition frame rate of camera 200 >=25 frame/seconds.
In the utility model one embodiment, data acquisition unit 300 can be implemented by hardware with analytical system 400.For hardware was implemented, embodiment described herein can realize in one or more special ICs (ASIC) or digital signal processor (DSP).In another embodiment of the utility model, data acquisition unit 300 can be made up by calculator with analytical system 400.
The embodiment of zebra fish behaviouristics system 1000 can be applicable to the zebra fish that grows up (back civilian abbreviation adult fish) OKR, adult fish OMR, zebra fish childhood (the back literary composition is called for short juvenile fish) OKR, the analysis of juvenile fish multi-point tracking widely.Yet in various embodiment, zebra fish behaviouristics system 1000 can be only applicable to the analysis of part occasion, and for example adult fish OKR analyzes.For the analytical system of implementing with hardware, in various embodiment, hardware possibly need change to some extent.
Fig. 2 illustrates the visual stimulus device sketch map of the utility model one embodiment.With reference to shown in Figure 2, this is exemplified as mechanical type visual stimulus device.The primary structure of zebra fish mechanical type visual stimulus device 100 comprises: housing 101, gear drive 102, motor 103, control panel 104, grating pallet 105, grating 106, annular light 107, fish jar pallet 108 and control circuit 110 etc.Motor 103 drives grating pallet 105 through gear drive 102 and rotates, and the rotation of grating pallet 105 drives grating 106 and rotates.Can control the switch of annular lamp 107, the rotating and the rotating speed of motor 103 through control panel 104.
Fig. 3 illustrates the control circuit of visual stimulus device shown in Figure 2, and with reference to shown in Figure 3, control circuit 110 is supplied power by the 220V power supply.Holes probe 111 is used to detect the real-time rotating speed of motor 103, and revolution counter 112 is used for display speed, and resistance R 1 is used to regulate the speed of motor.K switch 2, K3 are located on the control panel 104, control the rotating of annular lamp 107 and motor 103 respectively.
Mechanical type visual stimulus apparatus function and characteristics are:
1. uniform scattered light is provided, and raster image is evenly clear;
2. show the grating rotating speed in real time, and regulating grating rotation direction and grating rotating speed, to change grating be temporal frequency;
3. grating easy disassembling can be changed the spatial frequency of grating easily.
Except using the mechanical type visual stimulus device of above description, can also use the visual stimulus device 100 of another embodiment as shown in Figure 4, promptly arc screen stimulator is as the visual stimulus of zebra fish.Arc screen stimulator uses arc panel type display 128 to combine with fish jar or culture dish.Arc panel type display 128 can be realized multiple stimulus modality according to the programme-control display image.The grating that stimulates can change the direction of motion, color, angle and the asymmetric stimulation of zebra fish right and left eyes of spatial frequency, temporal frequency, grating arbitrarily, image that simultaneously also can show complex.
Fig. 5 illustrates the drive circuit of arc panel type display 128 shown in Figure 4.Comprise CPU122, memory 124, display interface 126 in the drive circuit.CPU 122 can be from memory 124 fetch program carry out, produce image, through the driving of display interface 126, on arc panel type display 128, show.
Owing to saved frame for movement, whole stimulating apparatus volume is little, simple for structure.And service routine control, realized automation, easy to use.
Fig. 6 illustrates the behaviouristics analytical system framework of the utility model one embodiment.With reference to shown in Figure 6, behaviouristics analytical system 400 can comprise that analysis module 402, data correction module 404 and data derive module 406.
In order to make this analytical system 400 be applicable to different situations such as adult fish OKR, adult fish OMR, juvenile fish OKR and the analysis of juvenile fish multi-point tracking; Above-mentioned these modules; Especially analysis module needs adjustment, to be used to analyze various zebra fish behaviouristics video data type.
Below be presented in the process of analyzing under the different occasions.
The OKR of adult fish
The OKR of adult fish carries out grating to stimulate under the condition that the zebra fish health is fixed, images of left and right eyes can be with the motion of following the tracks of grating.At this moment need analyze be in the video each frame images of left and right eyes with respect to the angle of center line movement.Obtain zebra fish OKR angle curve in time thus, can from curve, analyze the parameter such as angle, amplitude, cycle, number of times of eye movement.
At first be that the OKR behaviouristics is brought out.Grow up zebra fish after anaesthetic anesthesia, fix, stop it to move about with pin or other fixtures.Treat that the clear-headed back of zebra fish adapts to 5～10min, place mechanical type visual stimulus device (like Fig. 2) to fixing zebra fish, perhaps arc shields in the stimulator (like Fig. 4).Adapt to about 2min to light stimulus, through the OKR of the adult zebra fish head of camera 200 records, as shown in Figure 7.
Fig. 7 illustrates the adult zebra fish OKR analysis principle of the utility model one embodiment.Can find out from the morphological feature of adult fish shown in Figure 7: left eye of adult fish and right eye, dorsal part is seen, and can find out the characteristic curve of eyeball thus significantly.Therefore when analyzing; Can use certain part in the image as template; Use template matches (Match Pattern) algorithm to seek the image with template matches in the drawings, and the be complementary parameters such as number, direction, coordinate of image of final acquisition and left eye, right eye, center.
The analytic process of adult fish OKR is as shown in Figure 8.At first be the step S 1 that left eye, right eye, center die plates are set.Choosing of the template image that template matches (Pattern Match) the step S3 that this step is stated after realizing is required.Next is that ROI (region of interest, region of interest) is set, this step realize three with left eye, right eye, the corresponding area-of-interest in center.Carry out template matches at step S3, obtain left eye, right eye, corresponding coordinate and the angle of three templates in center through the image module matching algorithm.At last in data such as step S4 these coordinates of storage and angles.
Fig. 9 illustrates the adult zebra fish OKR assay surface 500 of the utility model one embodiment.There is image-region 502 in 500 left side at the interface, through three templates 504,506,508 being set as tracing object, in three ROI zones, passes through template matching algorithm respectively, follows the tracks of and obtains zebra fish eye movement angle-data in the video.On the right side at interface curve regions 510 is arranged, it shows the curve movement at left eye, right eye and center respectively.
The OMR of adult fish
The zebra fish that when grating moves, grows up can be followed grating and moved in a circle around fish jar, and this promptly is adult fish OMR.
At first adult fish is placed as in Fig. 2 or the visual stimulus device 100 shown in Figure 4, photopia is after a few minutes, and camera 200 record moving gratings are given the video of visual stimulus clockwise or counterclockwise.
Can know that through observing the motion of adult fish has following characteristics: in the space in the center of circle, make reciprocal circular motion; The bodily form can change in motion process.Therefore, be used in combination background correction and image binaryzation parser here,, isolate the target that to follow the tracks of through background deduction.After binaryzation, obtain the coordinate of two-value figure again.
Because zebra fish makes the circular motion of circulation, can from the figure of video different frame, merge through shearing, synthesize background.
The concrete analysis process is shown in figure 10.At first, use image subtracting background to be analyzed to obtain the object different with background in step S11 background correction.Because grating rotates, after the background correction, grating also can appear in the image.Therefore for fear of the interference of grating, at step S12, the ROI zone of in the image of background correction, shearing out a circle is as the object of analyzing; At step S13, threshold value is set image binaryzation, isolate target; The target that can obtain to follow the trail of through the binary map analysis of step S14 again; At last at step S15 storage data.
Figure 11 illustrates the adult zebra fish OMR assay surface 600 of the utility model one embodiment.In interface shown in Figure 11 600, there is image-region 602 in the left side, and the border circular areas that comprises the adult fish image 604 of its central authorities is the ROI zone.There is curve regions 606 on the right side, interface, and it shows the of living in angle change curve of adult fish in border circular areas 604.
Thus through synthetic background, background correction, ROI choose, binary-state threshold is provided with and obtains the target of following the tracks of; Analyze zebra fish in one section video contrary/number of turns of needle movement clockwise the time; And move distance, average speed, finally obtain the motion change after the zebra fish visual stimulus.
The OKR of juvenile fish
Because juvenile fish and adult fish are different on bodily form characteristic, so analytical method is corresponding adjusts.When analyzing juvenile fish OKR, defined axon as center line, the angle that right and left eyes rotates is reference with the center line, calculates its relative angle, and is shown in figure 12.The pigment of juvenile fish mainly concentrates on eye, also contains partial pigment at the pigment epithelial cell of head body surface, but more shallow with the eye color when compared, and disperses, and therefore through after the binaryzation, can from background, separate two eyeballs.
At first bring out.Juvenile fish is dipped in the methylcellulose, perhaps uses the low temperature agarose to fix its position, place juvenile fish in mechanical type visual stimulus device (like Fig. 2) or the arc screen stimulator (like Fig. 4), stimulate to grating, simultaneously camera 200 recording of video.
Analysis process is shown in figure 13.Flow process is perhaps successively carried out from both direction simultaneously.At step S21, each two field picture of video is at first chosen the reference center of a topography as juvenile fish eye movement angle from figure, be set to center die plates.At step S22, through template matching algorithm, obtain the coordinate and the angle of reference center, arrive step S26 storage data again.When analyzing the eye movement angle; At step S23; Choosing head is set is the ROI zone, then at step S24, after the post processing of extraction ROI image process binaryzation; It is 1 particle that two eyeballs correspond to two values, through can obtain left and right sides eyeball corresponding coordinate and angle in step S25 analysis of binary figure.Follow the tracks of in step S26 storage then and obtain data.
Figure 14 illustrates the zebra fish OKR assay surface 700 childhood of the utility model one embodiment.In interface shown in Figure 14 700; There is image-region 702 in the left side; The childhood vitellina is set as the center die plates of juvenile fish; The juvenile fish head zone 704 that a ROI rectangle frame defines is set, through obtaining the coordinate and the angle of right and left eyes after template matches, image binaryzation, the binaryzation analysis in image-region 702.There is curve regions 706 on the right side, interface, and it shows the angle change curve of juvenile fish at right and left eyes.
After video data was analyzed, in order to eliminate because of video image quality, parameter was provided with the profiling error of the few frames image that reason such as improper causes, and can these data manually be corrected, and obtains reliable data.
Be the data outputs at last, export the coordinate X of zebra fish, Y, totalframes, total time, total distance, average speed, total motion number of turns (OMR), the clockwise number of turns (OMR), the number of turns (OMR) counterclockwise at each frame; Eye movement angle (OKR), eye movement number of times (OKR), average eye movement amplitude (OKR).
Wherein, total time:
Total distance:
(i=N
Frames of viedo, i>=1), i is the video frame number, x, y are respectively the coordinate of template matches or binary map analysis acquisition.
Average speed:
The clockwise number of turns:
((A
i-A
I-1)>=0 or (A
i-A
I-1)≤180), (i=N
Frames ofviedo) when OMR analyzed, each coordinate that the binary map analysis obtains had an angle A with respect to the ROI centre coordinate, through contrasting in adjacent two two field pictures, the change of angle can be judged the direction that it moves about.The clockwise number of turns:
((A
i-A
I-1)＜0 or (A
i-A
I-1)＞180), (i=N
Framesof viedo).
Number of total coils: N=N
W+ N
AtW.
The moving angle of left eye is with respect to the angle at center: A
L-o=A
L-A
o
The moving angle of left eye is with respect to the angle at center: A
R-o=A
R-A
o
Wherein, A
L, A
R, A
oBe respectively the absolute angle at left eye, right eye, center, A
L-o, A
R-oBe respectively the angle of images of left and right eyes with respect to the center
Eye movement number of times: N
EyeThe crest number of=eye movement curve
Average eye movement amplitude:
Wherein j is total crest and trough number, Max
jAnd Min
jBe respectively crest and trough.
Though the utility model discloses as above with preferred embodiment; Right its is not in order to limit the utility model; Any those skilled in the art; In spirit that does not break away from the utility model and scope, when can doing a little modification and perfect, so the protection domain of the utility model is when being as the criterion with what claims defined.
Claims (3)
1. a zebra fish behaviouristics is brought out system, it is characterized in that comprising:
The visual stimulus device can bring out eye movement reaction, the optomotor response behavior of zebra fish at least;
Camera is in order to take the behavior of zebra fish, to obtain video information;
Data acquisition unit obtains video information from this camera, and imports an analytical system.
2. zebra fish behaviouristics as claimed in claim 1 is brought out system; It is characterized in that; Said visual stimulus device is a mechanical type visual stimulus device, and it comprises motor, transmission device, grating and fluorescent tube, and said motor is used to produce rotary power; Said transmission device is used for being passed to grating to the rotation of motor and rotates to drive grating, and said fluorescent tube is used to illuminate grating and produces visual pattern.
3. zebra fish behaviouristics as claimed in claim 1 is brought out system; It is characterized in that said visual stimulus device is an arc screen stimulator, it comprises container that holds zebra fish and the arc panel type display that surrounds this container; This arc panel type display produces image, as the visual stimulus of zebra fish.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104137800A (en) * | 2014-08-08 | 2014-11-12 | 青岛中仁药业有限公司 | Zebra fish electroconvulsive testing device |
| CN104542389A (en) * | 2014-12-23 | 2015-04-29 | 中国科学院苏州生物医学工程技术研究所 | Preparation method for non-alcoholic fatty liver disease zebra fish |
| CN114304005A (en) * | 2021-12-28 | 2022-04-12 | 中国科学院生物物理研究所 | Accurate visual stimulation's toy action recorder |
-
2011
- 2011-02-28 CN CN2011200497374U patent/CN202184048U/en not_active Expired - Lifetime
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104137800A (en) * | 2014-08-08 | 2014-11-12 | 青岛中仁药业有限公司 | Zebra fish electroconvulsive testing device |
| CN104137800B (en) * | 2014-08-08 | 2016-02-24 | 青岛中仁药业有限公司 | A kind of zebra fish electroshock seizure test device |
| CN104542389A (en) * | 2014-12-23 | 2015-04-29 | 中国科学院苏州生物医学工程技术研究所 | Preparation method for non-alcoholic fatty liver disease zebra fish |
| CN104542389B (en) * | 2014-12-23 | 2017-01-25 | 中国科学院苏州生物医学工程技术研究所 | Preparation method for non-alcoholic fatty liver disease zebra fish |
| CN114304005A (en) * | 2021-12-28 | 2022-04-12 | 中国科学院生物物理研究所 | Accurate visual stimulation's toy action recorder |
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