CN108902013A - A kind of method for building up of macular edema animal model - Google Patents
A kind of method for building up of macular edema animal model Download PDFInfo
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- CN108902013A CN108902013A CN201810547482.0A CN201810547482A CN108902013A CN 108902013 A CN108902013 A CN 108902013A CN 201810547482 A CN201810547482 A CN 201810547482A CN 108902013 A CN108902013 A CN 108902013A
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Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/02—Breeding vertebrates
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61D—VETERINARY INSTRUMENTS, IMPLEMENTS, TOOLS, OR METHODS
- A61D7/00—Devices or methods for introducing solid, liquid, or gaseous remedies or other materials into or onto the bodies of animals
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Abstract
The invention discloses a kind of method for building up of macular edema animal model.Using the reasonable application of 577nm laser joint Bengal rose red, block Rat retina vein, forms macular edema, use noninvasive OCTA/OCT technology continuous observation model eyes retina and its edematous state.This invention address that inventing, a kind of pair of animal pattern wound is small, the method for building up of rapid-action, length of holding time and simple and easy macular edema animal model, to meet corresponding requirement of experiment well.
Description
Technical field
The present invention relates to a kind of method for building up of macular edema animal model.
Background technique
Macular edema is more common in retinal vein obstruction, diabetic retinopathy as a kind of common Eye disease
After change, chronic uveitis, eye traumas and intraocular surgery etc..The conscious visual impairment of patient, metamorphopsia, are gradually aggravated, sternly
Ghost image rings patients ' life quality.The pathogenesis of macular edema is mainly since above-mentioned primary disease causes macular area capillary
Damage occurs leakage and causes oedema.There are a large amount of researchers to be dedicated to treating the medicament research and development of macular edema at present.It is preclinical
Phase experiment needs a kind of macular edema animal model, for observing the therapeutic effect of researched and developed drug.The foundation of the model is wanted
Ask simple, low wound, rapid-action, length of holding time.
According to the literature, the animal model that can first establish macular edema primary disease generates retina with progression of disease
Oedema.Diabetic retinopathy model foundation is time-consuming long, it is big to spend, and it is indefinite to generate macular edema probability；Chronic uvea
Scorching model involves anterior chamber of eye more, easily causes the complication such as cataract, macular edema formation probability is indefinite and observation is difficult；Eyeball
Macular edema caused by wound or intraocular surgery has certain contingency, non-artificial controllable.Above several schemes can not
The macular edema animal model that quick acting is provided, holds time long.It is contemplated that retinal vein could be blocked, cause
Macular edema model.
Retinal vein obstruction animal model has been reported that more.Preparation method includes that vascular ligation method, diathermy, fibrin ferment are quiet
Endothelin -1 method, laser photocoagulation and photochemical induction etc. are injected in arteries and veins instillation, vitreous chamber.Vascular ligation method：It is early
There is scholar to select macaque in 1978, is cut by lateral orbital wall, burns ligation central vein of retina, blood flow is caused to interrupt,
Distal blood flow backflow obstruction.This method is complicated for operation, and destructive larger, experimental animal is seriously wounded, postoperative easy accompanying infection, moves
The object death rate is high, superseded.Diathermy, i.e., coagulation retinal vein method in vitreum：Selection domestic pig is animal pattern, by two
Pole pipe probe is inserted into vitreous chamber at 3-4mm after corneoscleral junction, retinal vein above coagulation, until blood flow interrupts, is solidifying
It is stagnant.The complication such as the method also belongs to invasive surgical, and experimental animal wound is larger, and postoperative easy generation infection, intraocular pressure increase.It is solidifying
Hemase intravenous drip：Rabbit is selected, by operation on vitreous, fibrin ferment is directly instiled in retinal vein tube wall, 6 hours
Visible thrombosis occluding vascular afterwards.Time-consuming for this method occluding vascular, and easily causes other position blood vessels with blood flow operation
Obstruction.Vitreous chamber injects endothelin -1 method：Rabbit is selected to cause retinal vessel to receive endothelin -1 implantation glass body
Contracting, lumen reduce until entirely shutting.Endothelin -1 implantation dosage is indefinite in this method, causes blood vessel transience spasm or thorough
Bottom occlusion it is indefinite, and to the effect of retinal vessel be it is non-selective, obstructive position is unable to control.Laser photocoagulation：Select palm fibre
Color Norway (BN) rat or SD rat, laser irradiation retinal vessel, the thermal energy that laser generates make angiemphraxis.This method obstruction
Effect is of short duration, and experimental animal mostly revascularization occurs in 3 days or so after surgery.Photochemical induction：Select BN rat or SD rat,
Non-pigment/have the animals such as pigment rabbit, miniature pig combines photosensitizer using laser, such as chlorination thiosulfonic acid peptide cyanines, bengal rose
It is red, cause retinal vein obstruction.This method can cause the complication such as retinal hemorrhage, detachment of retina, and postoperative 3-5 days
See revascularization.There is scholar to prepare macular edema model by blocking retinal vein：SD rat is selected, is injected intravenously photosensitive
Agent Erythrosin B combines 532nm laser prolonged exposure retina 8 minutes, postoperative to occur injury of blood vessel and retina immediately
Thickness increases, but 5 days after operation retinal thickness restores to preoperative, and the obvious atrophy of generation retina in 14 days can not provide long-acting view
The animal model of nethike embrane oedema, and prolonged laser irradiation increases the damage to other positions of retina.
Therefore, it is necessary to establish a kind of animal model of macular edema, preparation method requires simple, low wound, rapid-action, dimension
It is long to hold the time.
Summary of the invention
In view of the problems of the existing technology, we pass through the study found that 577nm laser irradiation combines photosensitizer Bangladesh
The reasonable application of rose-red can block Rat retina vein and cause macular edema.This method is simple, low wound, and art finishes optics phase
Dry tomoscan blood flow imaging (OCTA) and optical coherence tomography (OCT) check to be visible macular edema, and can maintain
Up to 2 weeks.
Therefore, the purpose of the present invention is to provide a kind of novel methods for establishing macular edema animal model.The present invention adopts
With the reasonable application of 577nm laser irradiation joint Bengal rose red, use optical coherence tomography blood flow imaging (OCTA)
With optical coherence tomography (OCT) technology observation retinal morphology, blood flow and thickness.This invention address that establishing a kind of simple
It is easy, animal injury is small, oedema is rapid-action, long modeling method of holding time.The macular edema animal mould that the present invention establishes
Type can be very good to meet relevant requirement of experiment.
To achieve the above object, the present invention uses following technical scheme：
The first aspect of the invention provides a kind of 577nm laser irradiation joint photosensitizer Bengal rose red and is establishing
Application method in macular edema animal model.
The second aspect of the invention, provides a kind of method for building up of macular edema animal model, and this method includes：
(1) Bengal rose red solution is injected to the tail vein of experimental animal；And
(2) retinal vein of the 577nm laser irradiation experimental animal is used, to obtain the animal model.
The third aspect of the invention, provides a kind of method of observing and nursing animal retina oedema, and this method includes：It adopts
With optical coherence tomography blood flow imaging (OCTA) and optical coherence tomography (OCT) technology, thus observing and nursing animal
Retinal morphology, blood flow and thickness.
Compared with prior art, the present invention having the following advantages that：
Experimental animal wound is small, belongs to Noninvasive operation, and without complication such as postoperative infections；Accurate obstruction target blood
Pipe has high selectivity to retinal vessel, without other site tissue damages；Rapid occlusion target blood, in implementation process
Observe that distal vein backflow obstruction, proximal venal ischemic attenuate；Quick acting, art, which finishes, can be observed macular edema, water
It is swollen to maintain to provide grace time up to 2 weeks for the observation of related experiment.Macular edema is observed using OCTA/OCT technology,
Simply, noninvasive, favorable repeatability.
Detailed description of the invention
The Figure of description for constituting a part of the invention is used to provide further understanding of the present invention, and of the invention shows
Examples and descriptions thereof are used to explain the present invention for meaning property, does not constitute improper limitations of the present invention.
Fig. 1 .OCTA display model animal retina vein laser irradiation schematic diagram, target blood are grey mark (white
Shown in arrow).
In Fig. 2 embodiment of the present invention 1, normal BN rat is shot using OCTA technology Angio Retina 8.00mm mode
Surface layer retinal vascular morphologies (A), retinal morphology (B), retinal blood flow (C) and retinal thickness tomoscan result
(D)。
In Fig. 3 embodiment of the present invention 1, OCTA technology Angio Retina is used after A group rat laser irradiation immediately
8.00mm mode shoots retina.This figure is enumerated as 4 branches of laser irradiation retinal vein.
In Fig. 4 embodiment of the present invention 1, preoperative, postoperative 1 day, 2 weeks, the 3 weeks OCT inspection results of A group rat.
In Fig. 5 embodiment of the present invention 1, preoperative, postoperative 1 day, 2 weeks, the 3 weeks OCT inspection results of B group rat.
In Fig. 6 embodiment of the present invention 1, preoperative, postoperative 1 day, 2 weeks, the 3 weeks OCT inspection results of C group rat.
In Fig. 7 embodiment of the present invention 1, preoperative, postoperative 1 day, 1 week, the 2 weeks OCT inspection results of D group rat.
In Fig. 8 embodiment of the present invention 2, preoperative, postoperative 1 day, 2 weeks, the 3 weeks retinal tissue pathology results of A group mouse
(HE dyeing, 200 ×).
Specific embodiment
It is noted that described further below be all exemplary, it is intended to provide further instruction to the present invention.Unless another
It indicates, all technical and scientific terms used herein has usual with general technical staff of the technical field of the invention
The identical meanings of understanding.
It should be noted that term used herein above is merely to describe specific embodiment, and be not intended to restricted root
According to exemplary embodiments of the present invention.As used herein, unless the context clearly indicates otherwise, otherwise singular
Also it is intended to include plural form, additionally, it should be understood that, when in the present specification using term "comprising" and/or " packet
Include " when, indicate existing characteristics, step, operation and/or their combination.
In the present invention, PD refers to disc diameter (papillary diameter), the surveyors' staff as retinal surface.
As background technique is introduced, macular edema animal model is established in the prior art there are certain deficiency,
In order to solve technical problem as above, the present invention proposes a kind of method for building up of new macular edema animal model.
In one embodiment of the invention, it is related to a kind of 577nm laser irradiation joint photosensitizer Bengal rose red
In the application established in macular edema animal model.
In one embodiment of the invention, it is related to a kind of method for building up of macular edema animal model, this method
Including：
(1) Bengal rose red solution is injected to the tail vein of experimental animal；And
(2) retinal vein of the 577nm laser irradiation experimental animal is used, to obtain the animal model.
In one embodiment of the invention, the preparation method of the Bengal rose red solution includes：It is protected from light item
Under part, Bengal rose red powder is weighed, physiological saline is dissolved in, mixes well, Bengal rose red solution is made；Through
Stand-by Bengal rose red solution is obtained by filtration in sterilised membrane filter.
Preferably, the aperture of the sterilised membrane filter is 0.22 μm；After the completion of preparation, 4 DEG C be kept in dark place it is spare.
In one embodiment of the invention, experimental animal is BN rat or C57BL/6 mouse.
Preferably, the BN rat, female, 7-8 week old, weight 140-160g and the C57BL/6 mouse, it is female
Property, 6-8 week old, weight 18-22g.It is checked through slit-lamp microscope, OCTA/OCT before experiment, excludes experimental animal eyes and bend
The exception of light interstitial and retina.
In one embodiment of the invention, 10% (w/w) chloraldurate is injected intraperitoneally in experimental animal, after anesthesia is appropriate,
Tail vein injection Bengal rose red solution.
In one embodiment of the invention, after tail vein injection Bengal rose red solution 30-60 seconds, Ji Kejin
Row 577nm laser irradiation retinal vein.
The present invention follows low energy, small light spot, long time for exposure, the principle for repeating laser irradiation.At of the invention one
In embodiment, the 577nm laser irradiation parameters are：Spot diameter is 50-60 μm, and the time for exposure is 0.3 second, laser energy
For 60-80mW.By experimental verification, the optimal macular edema animal mould of effect can be obtained using the laser irradiation condition
Type.
In one embodiment of the invention, the specific method of the retinal vein of laser irradiation experimental animal includes：
After abundant mydriasis, targeted vein (Fig. 1) is determined within the scope of distance view nipple 1-3PD, from the near to the distant, to the vein for being about 1-2PD
Laser irradiation is carried out, until arriving distal vein backflow obstruction under the microscope, proximal venal ischemic attenuates.The present invention uses small light
Spot, the long time for exposure, repeats laser irradiation at low energy, stimulates irradiated Intravascular Thrombus to be formed, occluding vascular.Art finishes
Observe that retina thickens, postoperative 1 day retina diffusivity occurs and thickens.
In one embodiment of the invention, selection 2-3 branch retinal vein carries out laser irradiation, macular edema
Effect is best.
In one embodiment of the invention, the key of macular edema Animal Model is that obstruction retina is quiet
Arteries and veins observes venous thronbosis under slit-lamp microscope.Therewith using OCTA/OCT technology continuous observation retinal morphology,
Blood flow and retinal thickness.Animal is put to death in different time points, takes out eyeball, embedded section, and retina is observed in HE dyeing
Histological change.After retinal vein obstruction, OCTA/OCT inspection can may be immediately observed that retina thickens, postoperative 1 day view
Film is presented diffusivity and thickens, and maintains 2 weeks.Histological examination confirms OCTA/OCT inspection result.
In order to enable those skilled in the art can clearly understand technical solution of the present invention, below with reference to tool
The embodiment of the body technical solution that the present invention will be described in detail.
In embodiment, key instrument：French light too 577nm semiconductor laser；
Main agents：Bengal rose red powder is purchased from Sigma Co., USA；Compound tropicamide eye drops, hydrochloric acid are difficult to understand
Bucaine eye drops, Levofloxacin Eye drop are purchased from Japanese Santen Pharmaceutical Co. Ltd.；Physiological saline and 10% (w/w) water
Close chloral.
Embodiment 1：Macular edema model is established using BN rat
Step 1：The preparation of Bengal rose red solution
Under the conditions of being protected from light, precision weighing Bengal rose red powder 50mg is placed in 1.5ml brown EP pipe, is added
1.0ml physiological saline, mixes well.After 0.22 μm of sterilizing filter filter 23 time, Bengal rose red solution (50mg/ obtained
Ml), save backup for 4 DEG C.
Step 2：BN rat prepares
Female BN rat 4,7-8 week old, weight average 150g, 10% abdominal cavity (w/w) chloraldurate (0.3ml/100g)
Injection.Anaesthetize it is appropriate after, the abundant mydriasis of compound tropicamide eye drops, slit-lamp, OCTA/OCT check, record (Fig. 2).It is fixed
Rat, 75% alcohol of tail portion adequate disinfection.50mg/ml Bengal rose red solution (0.1ml/100g) is stored at room temperature 10 minutes
Row tail vein injection afterwards, injection finishes to be sterilized again, prevents from infecting.Oxybuprocaine hydrochloride eye drops row ocular facial anesthesia.Note
After penetrating Bengal rose red solution 30-60 seconds, laser irradiation can be started.
Step 3:577nm laser irradiation retinal vein
4 BN rats take right eye as model foundation eye, and left eye is control eye.It is respectively designated as A, B, C, D rat.
Retinal vessels in rats can be obviously observed under slit-lamp microscope, in the selection target vein within the scope of optic disk 1-3PD.
Follow low energy, small light spot, long time for exposure, the principle for repeating laser irradiation.Laser parameter is set：50-60 μm of spot diameter,
Time for exposure 0.3 second, laser energy 60-80mW.A mouse selects 4 branches of retinal vein：Upper and lower, left and right；B mouse selection 3
A branch；C mouse selects 2 branches；D mouse selects 1 branch.It is repeated as many times laser irradiation from the near to the distant apart from optic disk, closes quiet
Arteries and veins 1-2PD length is formed until observing that distal vein backflow obstruction, proximal venal ischemic attenuate under slit-lamp microscope
Retinal vein obstruction.Art, which finishes, gives Levofloxacin Eye drop eye droppings.
Step 4：Observing and nursing animal retina edematous condition
After observing retinal vein obstruction under slit-lamp microscope, row OCTA checks (Fig. 3) immediately.After laser operation daily
Slit lamp observation art eye prosthomere observes retina using OCTA/OCT technology, until macular edema subsides.It records and compares every
The eyes situation of group rat and different group rat model eye situations.
Step 5：Interpretation of result
Every group of BN rat occluding vascular quantity is different, and view film reaction is different.It is observable after A Rat retina vein obstruction
To retinal blood flow signal interruption or missing, macular edema is thickened, and postoperative 1 day retina diffusivity thickens, neural epithelium is de-
From retina obviously thickens within more postoperative 1 day postoperative 2 weeks, and slight neural epithelium is detached from, postoperative 3 weeks macular edemas mitigation, structure
Disorder (Fig. 4).B, the postoperative 1 day retina diffusivity oedema of C mouse, postoperative 2 weeks still visible retina diffusivity oedema, postoperative 3 weeks
Macular edema recession, obvious thin change (Fig. 5, Fig. 6).The postoperative 1 day retina diffusivity of D mouse thickens, postoperative 1 week macular edema
Substantially reduced, postoperative 2 weeks macular edemas subside completely, slight thin change (Fig. 7).
It summarizes：Photosensitizer Bengal rose red is combined using 577nm laser, laser is using low energy, small light spot, long exposure
Time selects 2-3 branch vein, and reirradiation, oedema formation effect is best, holds time up to 2 weeks, and to retinal damage
It is small, meet the requirement to macular edema animal model.Because the experimental animal that retina correlative study is often used includes BN rat
With C57BL/6 mouse, therefore select C57BL/6 mouse verifying.
Embodiment 2：Macular edema model is established using C57BL/6 mouse
Step 1：The preparation of Bengal rose red solution
Under the conditions of being protected from light, precision weighing Bengal rose red powder 25mg is placed in 1.5ml brown EP pipe, is added
1.0ml physiological saline, mixes well.After 0.22 μm of sterilizing filter filter 23 time, Bengal rose red solution (25mg/ is made
Ml), save backup for 4 DEG C.
Step 2：C57BL/6 mouse prepares
Female C57BL/6 mouse 15,6-8 week old, weight average 20g, 10% (w/w) chloraldurate (1ml/100g) abdomen
Chamber injection, after anesthesia is appropriate, the abundant mydriasis of compound tropicamide eye drops, slit-lamp, OCTA and OCT are checked, record.It is fixed
Mouse, 75% alcohol of tail portion adequate disinfection.25mg/ml bengal rose solution (0.5ml/100g) is stored at room temperature after ten minutes
Row tail vein injection, injection finish alcohol disinfecting again, prevent from infecting.Oxybuprocaine hydrochloride eye drops row surface anesthesia.Note
After penetrating Bengal rose red solution 30-60 seconds, laser irradiation can be started.
Step 3:577nm laser irradiation retinal vein
15 mouse are randomly divided into tri- groups of A, B, C, every group 5.Three groups of mouse take right eye as model foundation eye, left eye
To compare eye.It is respectively designated as A, B, C group mouse.Mouse Retina blood vessel can be obviously observed under slit-lamp microscope, managed
Diameter is thin compared with rat aorta.Low energy, small light spot, long time for exposure are followed, the principle of laser irradiation is repeated, laser parameter is set：
50 μm of spot diameter, the time for exposure 0.3 second, laser energy 60mW.In the selection target vein within the scope of optic disk 1-3PD.
A group mouse selects 3 veins, and B group mouse selects 2 veins, and C group mouse selects 1 vein.It is repeated from the near to the distant apart from optic disk
Laser irradiation vein 1-2PD length, until observing distal vein backflow obstruction, proximal venal ischemic under slit-lamp microscope
Attenuate, forms retinal vein obstruction.Art, which finishes, gives Levofloxacin Eye drop eye droppings.
Step 4：Observing and nursing animal retina edematous condition
After observing retinal vein obstruction under slit-lamp microscope, row OCTA is checked immediately.Daily slit-lamp after laser operation
Art eye prosthomere is observed, observes retinal thickness using OCT technology, until macular edema subsides.Before laser operation, it is 1 day postoperative,
Eyeball of mouse, embedded section, HE dyeing, observation retinal structure variation are taken within 1 week, 2 weeks and 3 weeks respectively.
Step 5：Interpretation of result
Every group of mouse occluding vascular quantity is different, and view film reaction is different.OCT inspection result shows macular edema degree
And it holds time：A group > B group > C group.
Every group of mouse respectively before laser operation, take within postoperative 1 day, 1 week, 2 weeks and 3 weeks eyeball of mouse, embedded section, HE dye
Color, each layer structure of retina changes as the result is shown, especially obvious with inner molecular layer, inner nuclear layer variation.Each group Mouse Retina
Changes in microstructure result meets OCT observation acquired results.By taking A group mouse as an example：Postoperative 1 day inner plexiform layer of retina thickens,
Inner nuclear layer cellular swelling, postoperative 2 weeks still visible inner plexiform layer of retina thicken, inner nuclear layer eucaryotic cell structure disorder, postoperative 3 weeks views
Film inner molecular layer edema extinction, inner nuclear layer is slight thin change (Fig. 8)
It summarizes：BN rat result in C57BL/6 mouse results verifying embodiment 1 in embodiment 2.Follow low energy, small light
Spot, long time for exposure, the principle for repeating laser irradiation combine photosensitizer Bengal rose red using 577nm laser, block 2-3
Branch retinal vein, oedema formation effect is best, holds time up to 2 weeks, small to retinal damage, meets to macular edema
The requirement of animal model.
The above embodiment is a preferred embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, other any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present invention,
It should be equivalent substitute mode, be included within the scope of the present invention.
Claims (10)
1. a kind of 577nm laser irradiation joint photosensitizer Bengal rose red is establishing answering in macular edema animal model
With.
2. a kind of method for building up of macular edema animal model, characterized in that this method includes：
(1) Bengal rose red solution is injected to experimental animal tail vein；And
(2) retinal vein of the 577nm laser irradiation experimental animal is used, to obtain the animal model.
3. method for building up as claimed in claim 2, it is characterized in that：The 577nm laser irradiation parameters are：Spot diameter is
50-60 μm, the time for exposure is 0.3 second, laser energy 60-80mW.
4. method for building up as claimed in claim 2, characterized in that the specific side of the retinal vein of laser irradiation experimental animal
Method includes：Targeted retinal vein is determined within the scope of distance view nipple 1-3PD, by closely to remote, being repeated as many times laser along vein
Irradiation, the long 1-2PD of sealing blood vessels, until observing that distal vein backflow obstruction, proximal venal ischemic attenuate.
5. method for building up as claimed in claim 2, it is characterized in that：2-3 branch retinal vein is selected to carry out laser irradiation.
6. method for building up as claimed in claim 2, it is characterized in that：Experimental animal is BN rat or C57BL/6 mouse；It is preferred that
, the BN rat is female, 7-8 week old, weight 140-160g and the C57BL/6 mouse, female, 6-8 week old, weight
18-22g；
Preferably, it is checked through slit-lamp microscope and OCTA/OCT before experiment, excludes eyes refractive media or retina is different
Often, experimental animal tail portion alcohol disinfecting, it is spare.
7. method for building up as claimed in claim 2, it is characterized in that：10% (w/w) chloraldurate fiber crops are injected intraperitoneally in experimental animal
It is liquor-saturated it is appropriate after, tail vein injection Bengal rose red solution.
8. method for building up as claimed in claim 2, it is characterized in that：After injection Bengal rose red solution 30-60 seconds, Ji Kejin
Row 577nm laser irradiation retinal vein.
9. method for building up as claimed in claim 2, characterized in that the preparation method packet of the Bengal rose red solution
It includes：Under the conditions of being protected from light, Bengal rose red powder is weighed, physiological saline is dissolved in, fullys shake, shake up, through sterile filter
Bengal rose red solution is obtained by filtration in film；
Preferably, the aperture of the sterilised membrane filter is 0.22 μm；After the completion of preparation, 4 DEG C are protected from light, and are saved backup.
10. a kind of method for observing animal pattern macular edema described in claim 1, this method include：Using optics phase
Dry tomoscan blood flow imaging (OCTA) and optical coherence tomography (OCT) technology, thus observing and nursing animal retina shape
State, blood flow and thickness.
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