CN103743895A - Detachable glass slide incubator - Google Patents
Detachable glass slide incubator Download PDFInfo
- Publication number
- CN103743895A CN103743895A CN201410008291.9A CN201410008291A CN103743895A CN 103743895 A CN103743895 A CN 103743895A CN 201410008291 A CN201410008291 A CN 201410008291A CN 103743895 A CN103743895 A CN 103743895A
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- Prior art keywords
- microslide
- frame
- resilient clamp
- hatch
- stainless steel
- Prior art date
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- 239000011521 glass Substances 0.000 title abstract 9
- 229910001220 stainless steel Inorganic materials 0.000 claims abstract description 31
- 239000010935 stainless steel Substances 0.000 claims abstract description 31
- 238000000034 method Methods 0.000 claims abstract description 28
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 21
- 239000000741 silica gel Substances 0.000 claims abstract description 21
- 229910002027 silica gel Inorganic materials 0.000 claims abstract description 21
- 230000012447 hatching Effects 0.000 claims description 24
- 229960001866 silicon dioxide Drugs 0.000 claims description 19
- 230000002093 peripheral effect Effects 0.000 claims description 9
- 238000005406 washing Methods 0.000 claims description 8
- 238000011534 incubation Methods 0.000 abstract description 30
- 239000007788 liquid Substances 0.000 abstract description 16
- 238000011532 immunohistochemical staining Methods 0.000 abstract description 6
- 238000002474 experimental method Methods 0.000 description 10
- 230000018044 dehydration Effects 0.000 description 6
- 238000006297 dehydration reaction Methods 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 5
- 238000003364 immunohistochemistry Methods 0.000 description 5
- 239000000463 material Substances 0.000 description 5
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- 238000007789 sealing Methods 0.000 description 4
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 238000005034 decoration Methods 0.000 description 2
- 240000008067 Cucumis sativus Species 0.000 description 1
- 235000010799 Cucumis sativus var sativus Nutrition 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 230000001744 histochemical effect Effects 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 230000002055 immunohistochemical effect Effects 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 230000009870 specific binding Effects 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
Images
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/5302—Apparatus specially adapted for immunological test procedures
- G01N33/5304—Reaction vessels, e.g. agglutination plates
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
- G01N1/31—Apparatus therefor
Abstract
The invention discloses a detachable glass slide incubator. The detachable glass slide incubator comprises a stainless steel incubation frame, a silica gel cushion layer, and elastic clampers which clamps the stainless steel incubation frame and the silica gel cushion layer on a glass slide, wherein the elastic clampers clamp the incubation frame and the glass slide through clamping openings of the elastic clampers; the incubation frame and the glass slide are clamped together to form a small incubation chamber through clamping parts of the elastic clampers. The detachable glass slide incubator is reasonable in structure; when immunohistochemical staining of a tissue is performed, a non-leakage incubation small chamber is formed by tightly pressing the incubation frame on the glass slide to which the tissue sections adhere by the elastic clampers repeatedly; when anti-body incubation is performed, only a trace amount of anti-body incubation liquid can ensure that all the tissue sections adhered to the glass slide are completely soaked in the anti-body incubation liquid in the process of incubating, and thus the experimental cost for scientific research is saved; other experimental steps can also be prevented from being influenced by the incubation frame by detaching the incubation frame.
Description
Technical field
The present invention relates to a kind of dismountable microslide couveuse.
Background technology
In carrying out the research of life science, people often need to use high specificity, highly sensitive immunohistochemistry technology to position histiocytic predetermined substance and quantitatively.The ultimate principle of immunohistochemistry technology is first to utilize immunologic known antibodies (first antibody) antigen in tissues observed to be combined.Because the antigen-antibody reaction of carrying out on tissue and cell is generally sightless, need to certain label (as enzyme, fluorescein) be attached on first antibody by the method for mark, Zai Yong histochemical method shows this label or the fluorescence sending at the micro-Microscopic observation fluorescein of fluorescent microscope, lazer scan confocal microscope or ultrahigh resolution.Accomplish the Cucumber in research object tissue visual.When stating in realization the method with mark of mentioning certain label (as enzyme, fluorescein) being attached on first antibody, people often adopt the method in conjunction with the second antibody of upper certain label and the specific binding of first antibody.
Yong immunohistochemistry technology positions histiocytic predetermined substance and quantitatively time, and people often adopt paster decoration method to keep the front and back order of the good form of histotomy and clear and definite each histotomy in experiment.The ultimate principle of paster decoration method is on microtome, to cut after tissue, immediately this histotomy is sequentially sticked on microslide by section.The experimental implementation such as follow-up sealing, the first antibody of histotomy hatched, washing, second antibody are hatched, wash again, dehydration, transparent and mounting are all to carry out under this histotomy sticks on the condition of microslide.
In sealing, the washing of above-mentioned histotomy, wash again, in dehydration, transparent and mounting process, because the relevant reagent cost of using is lower, in addition in washing process, the large usage quantity of cleansing solution.In order to obtain good experiment effect, people often adopt the method that the whole microslide that is pasted with histotomy is placed in above-mentioned relevant liquid to process.But carrying out first antibody and second antibody while hatching, because antibody price is higher, (price of at present general monoclonal first antibody 100 μ l is 3000 yuan of left and right, the price of special monoclonal first antibody can be higher), and attenuable multiple can not be very large while using.Therefore, antibody especially first antibody be the major part of whole experimental cost.So Yong immunohistochemistry technology positions histiocytic predetermined substance and quantitatively time, can not adopt the method that the whole microslide that is pasted with histotomy is placed in antibody incubation liquid to complete.
When carrying out immunohistochemical staining with paster method, people conventionally adopt antibody incubation liquid are directly added in to the method on the histotomy that microslide adheres to.But while using this method, antibody incubation liquid has been added can preserve and diffuse out microslide loss, add lacked the antibody incubation liquid on histotomy can be in the process of hatching, because moisture evaporation in Incubating Solution can not keep the histotomy on microslide to be immersed in all the time in antibody incubation liquid.Sometimes, people also can use the histotomy attaching at microslide with oily material, to draw the method for a frame around, and add antibody incubation liquid in this oiliness picture frame, prevent overflowing of antibody incubation liquid.Although but the method improves to some extent than method antibody incubation liquid being directly added on the histotomy that microslide attaches, but use the method, because oiliness picture frame stops the excessive poor ability of antibody incubation liquid, can not guarantee that histotomy is immersed in antibody incubation liquid all the time in whole antibody incubation process.And in antibody incubation, washing, dehydration, transparent process, these oily materials can come off and lose the excessive effect of antibody incubation liquid stoping in antibody incubation process; And oily material can occur come off and stick on histotomy, affect the situation that histotomy is observed; When oily material also can dissolve in relevant experiment reagent in histotomy is processed, the recycling of experiment reagent and the treatment effect of histotomy that because polluting experiment reagent, affect.
Summary of the invention
The object of the present invention is to provide experimental procedure energy convenience and high-efficiency a kind of rational in infrastructure, all to carry out, can obtain dismountable microslide couveuse of the experimental result of good immunohistochemical staining.
Technical solution of the present invention is:
A dismountable microslide couveuse, is characterized in that: comprise that stainless steel hatches frame, stainless steel is hatched frame and is pressed on microslide, jointly by middle coffin, take microslide and forms and hatch cell as bottom surface with the silica gel bed course below it; Described silica gel bed course has with stainless steel hatches the intermediate rectangular space that frame is identical, and sticks to stainless steel and hatch on frame, contacts with microslide, prevents the leakage of Incubating Solution; Stainless steel hatches the peripheral dimension of frame and the peripheral dimension of silica gel bed course is all identical with microslide peripheral dimension; Separately be provided with and stainless steel hatched to frame and silica gel bed course is clamped in the resilient clamp on microslide; Described resilient clamp comprises the clamping part of resilient clamp root, resilient clamp and the clamp port of resilient clamp; Resilient clamp root connects resilient clamp clamping part, and the hand position of resilient clamp when forming microslide couveuse and the operation of dismounting microslide couveuse is provided; Clamp port by resilient clamp is clamped in resilient clamp to hatch on frame and microslide, and the clamping part of resilient clamp will be hatched frame and microslide and is clamped together to form and hatch cell.
Its using method is: stainless steel is hatched to frame and silicagel pad is laminated on the microslide that is pasted with histotomy, guarantee that all histotomies all hatch cell bottom what hatch that frame and microslide form; Select stainless steel to hatch any one side in frame and upper and lower overlapping four limits of microslide, hand-held resilient clamp root, the clamp port of resilient clamp is aimed at and hatched the structure that frame and microslide are stacked up and down, resilient clamp is pushed and hatches the structure that frame and microslide are stacked up and down, the clamping part that guarantees resilient clamp will be hatched together with frame is pressed on microslide, its excess-three limit is repeated above-mentioned action resilient clamp is sandwiched, guarantee to hatch frame and be closely connected with microslide, form the cell of hatching not leaking; To hatching in cell, add first antibody Incubating Solution, guarantee can be immersed in first antibody Incubating Solution at the histotomy of hatching cell bottom, carry out on request hatching of histotomy; After having hatched for the first time, take out containing special first antibody Incubating Solution, unload and hatch frame and the microslide resilient clamp on phase stack structure four limits up and down, under microslide is dismantled, hatch frame the histotomy sticking on microslide is carried out to carrying out washing treatment; Carry out second antibody while hatching, repeat above-mentioned action.
The present invention is rational in infrastructure, when the immunohistochemical staining of organizing, can be by repeatedly using resilient clamp to hatch frame being pressed on the microslide that is pasted with histotomy tightly, form one do not leak hatch cell.When carrying out antibody incubation, only use micro-antibody incubation liquid, just guaranteed to stick on all histotomies on microslide in the process of hatching, be fully immersed in antibody incubation liquid, saved the experimental cost of scientific research.Meanwhile, by repeatedly unloading, hatch frame, guaranteed again microslide pre-treatment, organize paster, the experimental procedure such as washing, dehydration, transparent, mounting can not hatched the impact of frame, and avoided at histotomy around by the shortcoming of oily material picture frame.All experimental procedure energy convenience and high-efficiencies are carried out, guarantee finally can obtain the experimental result of a good immunohistochemical staining.
Accompanying drawing explanation
Below in conjunction with drawings and Examples, the invention will be further described.
Fig. 1 be dismountable microslide couveuse hatch frame schematic perspective view.
Fig. 2 is the microslide schematic perspective view of dismountable microslide couveuse.
Fig. 3 is the resilient clamp schematic perspective view of dismountable microslide couveuse.
Fig. 4 is dismountable microslide couveuse longitudinal profile schematic diagram.
In figure: 1. stainless steel is hatched frame: be pressed on microslide, jointly take microslide by middle coffin and form and hatch cell as bottom surface with the silica gel bed course below it.
2. stainless steel is hatched the silica gel bed course of frame: have with stainless steel and hatch the intermediate rectangular space that frame is identical, and stick to stainless steel and hatch on frame, contact with microslide, prevent the leakage of Incubating Solution.
3. microslide: carrying histotomy, and as the bottom surface of hatching cell.
4. resilient clamp root: connect resilient clamp clamping part, and the hand position of resilient clamp when forming microslide couveuse and the operation of dismounting microslide couveuse is provided.
5. the clamping part of resilient clamp: will hatch frame and microslide and be clamped together to form and hatch cell.
6. the clamp port of resilient clamp: resilient clamp is clamped in and is hatched on frame and microslide by this clamp port.
7. hatch cell: form small space and deposit Incubating Solution, accomplish both to use micro-Incubating Solution, guaranteed that again the histotomy on microslide is immersed in Incubating Solution all the time.
8. resilient clamp.
Embodiment
A dismountable microslide couveuse, is characterized in that: comprise that stainless steel hatches frame, stainless steel is hatched frame and is pressed on microslide, jointly by middle coffin, take microslide and forms and hatch cell as bottom surface with the silica gel bed course below it; Described silica gel bed course has with stainless steel hatches the intermediate rectangular space that frame is identical, and sticks to stainless steel and hatch on frame, contacts with microslide, prevents the leakage of Incubating Solution; Stainless steel hatches the peripheral dimension of frame and the peripheral dimension of silica gel bed course is all identical with microslide peripheral dimension; Separately be provided with and stainless steel hatched to frame and silica gel bed course is clamped in the resilient clamp on microslide; Described resilient clamp comprises the clamping part of resilient clamp root, resilient clamp and the clamp port of resilient clamp; Resilient clamp root connects resilient clamp clamping part, and the hand position of resilient clamp when forming microslide couveuse and the operation of dismounting microslide couveuse is provided; Clamp port by resilient clamp is clamped in resilient clamp to hatch on frame and microslide, and the clamping part of resilient clamp will be hatched frame and microslide and is clamped together to form and hatch cell.
Its using method is: according to immunohistochemical requirement, process the microslide of selecting, to guarantee that histotomy has good sticking effect on microslide.Choose the required tissue of experiment and cut into slices, and be attached on microslide.The sealing that the histotomy being attached on microslide is carried out before antibody incubation is processed.Complete after above-mentioned experimental procedure, stainless steel is hatched to frame (1) and silica gel bed course (2) and be pressed in that to be pasted with the microslide (3) of histotomy upper, guarantee that all histotomies are all hatching bottom cell (7) of hatching that frame and microslide form.Any one side in frame and upper and lower overlapping four limits of microslide is hatched in selection, hand-held resilient clamp root (4), the clamp port of resilient clamp (6) is aimed at and to be hatched the structure that frame and microslide are stacked up and down, resilient clamp is pushed and hatches the structure that frame and microslide are stacked up and down.The clamping part (5) that guarantees resilient clamp will be hatched frame and microslide forcing together tightly.Its excess-three limit is repeated above-mentioned action resilient clamp (8) is sandwiched.Guarantee to hatch frame and microslide close contact, form the cell of hatching not leaking.To hatching, in cell, add containing special first antibody Incubating Solution, guarantee to stick on the histotomy of hatching cell bottom and can be immersed in containing in special first antibody Incubating Solution.Carry out on request hatching of histotomy.After having hatched for the first time, take out containing special first antibody Incubating Solution, unload and hatch frame and the microslide resilient clamp on phase stack structure four limits up and down, from microslide dismounting, descend stainless steel to hatch frame and silica gel bed course.Routinely the histotomy sticking on microslide is carried out to carrying out washing treatment.Need to carry out second antibody while hatching, repeat above-mentioned action, again form the cell of hatching not leaking, add containing special second antibody Incubating Solution, histotomy is hatched.After having hatched, remove containing special second antibody Incubating Solution, the lower stainless steel of dismounting is hatched frame and silica gel bed course, washs, the aftertreatment such as dehydration, transparent, mounting to sticking on histotomy on microslide.After all finishing dealing with, examine under a microscope the immunohistochemical staining result of histotomy.
Use dismountable microslide couveuse of the present invention, first do not install and hatch frame, the pre-service of conveniently carrying out for microslide paster effect, and guarantee when carrying out histotomy, whole slide surface is smooth, good paster and the histotomy sealing effect of histotomy after cutting.Carrying out first antibody while hatching, on microslide, installing and hatch frame, form one do not leak hatch cell, use micro-first antibody Incubating Solution, all histotomies that just can make to stick on microslide are fully immersed in first antibody Incubating Solution.When first antibody has been hatched, from microslide, pull down and hatch after frame, the histotomy sticking on microslide is fully washed.Washed and carried out second antibody while hatching, on microslide, again install and hatch frame, form again one do not leak hatch cell, use equally micro-second antibody Incubating Solution, all histotomies that just can make to stick on microslide are fully immersed in second antibody Incubating Solution.When second antibody has been hatched, again from microslide, pull down and hatch after frame, to sticking on histotomy on microslide, again wash fully, dehydration, transparent and mounting process.To obtain good immunohistochemistry result.
Claims (2)
1. a dismountable microslide couveuse, is characterized in that: comprise that stainless steel hatches frame, stainless steel is hatched frame and is pressed on microslide, jointly by middle coffin, take microslide and forms and hatch cell as bottom surface with the silica gel bed course below it; Described silica gel bed course has with stainless steel hatches the intermediate rectangular space that frame is identical, and sticks to stainless steel and hatch on frame, contacts with microslide, prevents the leakage of Incubating Solution; Stainless steel hatches the peripheral dimension of frame and the peripheral dimension of silica gel bed course is all identical with microslide peripheral dimension; Separately be provided with and stainless steel hatched to frame and silica gel bed course is clamped in the resilient clamp on microslide; Described resilient clamp comprises the clamping part of resilient clamp root, resilient clamp and the clamp port of resilient clamp; Resilient clamp root connects resilient clamp clamping part, and the hand position of resilient clamp when forming microslide couveuse and the operation of dismounting microslide couveuse is provided; Clamp port by resilient clamp is clamped in resilient clamp to hatch on frame and microslide, and the clamping part of resilient clamp will be hatched frame and microslide and is clamped together to form and hatch cell.
2. dismountable microslide couveuse according to claim 1, it is characterized in that: its using method is: stainless steel is hatched to frame and silicagel pad is laminated on the microslide that is pasted with histotomy, guarantee that all histotomies all hatch cell bottom what hatch that frame and microslide form; Select stainless steel to hatch any one side in frame and upper and lower overlapping four limits of microslide, hand-held resilient clamp root, the clamp port of resilient clamp is aimed at and hatched the structure that frame and microslide are stacked up and down, resilient clamp is pushed and hatches the structure that frame and microslide are stacked up and down, the clamping part that guarantees resilient clamp will be hatched together with frame is pressed on microslide, its excess-three limit is repeated above-mentioned action resilient clamp is sandwiched, guarantee to hatch frame and be closely connected with microslide, form the cell of hatching not leaking; To hatching in cell, add first antibody Incubating Solution, guarantee can be immersed in first antibody Incubating Solution at the histotomy of hatching cell bottom, carry out on request hatching of histotomy; After having hatched for the first time, take out containing special first antibody Incubating Solution, unload and hatch frame and the microslide resilient clamp on phase stack structure four limits up and down, under microslide is dismantled, hatch frame the histotomy sticking on microslide is carried out to carrying out washing treatment; Carry out second antibody while hatching, repeat above-mentioned action.
Priority Applications (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510091595.0A CN104614517B (en) | 2014-01-09 | 2014-01-09 | The using method of dismountable microslide couveuse |
| CN201510091592.7A CN104614514B (en) | 2014-01-09 | 2014-01-09 | Dismountable microslide couveuse that experimental procedure energy convenience and high-efficiency carries out |
| CN201410008291.9A CN103743895B (en) | 2014-01-09 | 2014-01-09 | Detachable glass slide incubator |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410008291.9A CN103743895B (en) | 2014-01-09 | 2014-01-09 | Detachable glass slide incubator |
Related Child Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510091595.0A Division CN104614517B (en) | 2014-01-09 | 2014-01-09 | The using method of dismountable microslide couveuse |
| CN201510091592.7A Division CN104614514B (en) | 2014-01-09 | 2014-01-09 | Dismountable microslide couveuse that experimental procedure energy convenience and high-efficiency carries out |
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| Publication Number | Publication Date |
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| CN103743895A true CN103743895A (en) | 2014-04-23 |
| CN103743895B CN103743895B (en) | 2015-05-27 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201410008291.9A Expired - Fee Related CN103743895B (en) | 2014-01-09 | 2014-01-09 | Detachable glass slide incubator |
| CN201510091595.0A Expired - Fee Related CN104614517B (en) | 2014-01-09 | 2014-01-09 | The using method of dismountable microslide couveuse |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201510091595.0A Expired - Fee Related CN104614517B (en) | 2014-01-09 | 2014-01-09 | The using method of dismountable microslide couveuse |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104614514A (en) * | 2014-01-09 | 2015-05-13 | 南通大学 | Detachable slide incubator ensuring experimental steps to be carried out conveniently and efficiently |
| WO2021068560A1 (en) * | 2020-06-28 | 2021-04-15 | 南通大学 | Convenient-assembly, detachable incubator for microscope slide |
| WO2021073317A1 (en) * | 2020-09-09 | 2021-04-22 | 南通大学 | Method for using anti-evaporation detachable-type slide incubator |
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| WO2021073317A1 (en) * | 2020-09-09 | 2021-04-22 | 南通大学 | Method for using anti-evaporation detachable-type slide incubator |
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| Publication number | Publication date |
|---|---|
| CN104614517A (en) | 2015-05-13 |
| CN104614517B (en) | 2016-03-02 |
| CN103743895B (en) | 2015-05-27 |
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