CN105424449A - Flow-guiding body structure used for section staining and section staining method - Google Patents
Flow-guiding body structure used for section staining and section staining method Download PDFInfo
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- CN105424449A CN105424449A CN201510981522.9A CN201510981522A CN105424449A CN 105424449 A CN105424449 A CN 105424449A CN 201510981522 A CN201510981522 A CN 201510981522A CN 105424449 A CN105424449 A CN 105424449A
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- 238000010186 staining Methods 0.000 title claims abstract description 14
- 238000007447 staining method Methods 0.000 title 1
- 239000000975 dye Substances 0.000 claims description 101
- 239000000523 sample Substances 0.000 claims description 43
- 239000012530 fluid Substances 0.000 claims description 29
- 238000004043 dyeing Methods 0.000 claims description 28
- 238000000034 method Methods 0.000 claims description 21
- 230000008676 import Effects 0.000 claims description 9
- 239000012472 biological sample Substances 0.000 claims description 7
- 230000008859 change Effects 0.000 claims description 6
- 239000011521 glass Substances 0.000 claims description 4
- 239000007769 metal material Substances 0.000 claims description 4
- 239000007788 liquid Substances 0.000 abstract 6
- 238000002791 soaking Methods 0.000 description 5
- 230000003068 static effect Effects 0.000 description 4
- 230000008569 process Effects 0.000 description 3
- 230000003139 buffering effect Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000008520 organization Effects 0.000 description 2
- 230000009471 action Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 238000004040 coloring Methods 0.000 description 1
- 238000010205 computational analysis Methods 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000009795 derivation Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 238000006073 displacement reaction Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
Classifications
-
- 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
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Sampling And Sample Adjustment (AREA)
Abstract
The invention discloses a flow-guiding body structure used for section staining, which is characterized by including a flow-guiding main body (1). A flat flowing trough (2) is arranged on the flow-guiding main body (1). A liquid inlet buffer trough (3) is disposed on one end of the flat flowing trough (2) and is communicated with the flat flowing trough (2). A liquid outlet buffer trough (5) is disposed on the other end of the flat flowing trough (2) and is communicated with the flat flowing trough (2). A liquid inlet hole (4) is formed in the side wall of the liquid inlet buffer trough (3). A liquid outlet hole (6) is formed in the side wall of the liquid outlet buffer trough (6). The depth of the flat flowing trough (2) is 150-250 [mu]m. The flow-guiding body structure is simple and is low in cost. A stable flow field can be formed in the flat flowing trough through a dye for performing staining, thereby reducing false positive rate. Meanwhile, the flow-guiding body structure can reduce use amount of the dye, can increase staining accuracy and is suitable for being promoted.
Description
Technical field
The present invention relates to a kind of biological sample disposal route, particularly relate to a kind of baffle structure for section statining and section statining method.
Background technology
Section statining is the basic means of biological study, the process of section statining biological sample is cut into the thin slice that thickness is 5-40 microns, then by slice sticker on microslide, and making the different constituents of biological sample show different colors by dyeing, biology carrys out the Content and distribution of the protein ingredient of computational analysis particular organization by the size and the depth analyzing the biological sample particular organization color range after dyeing.At present, section statining adopts the method for soaking and dyeing usually, is specially and is cut into slices by sample, then according to set order, soak successively in different dyestuffs, by controlling the soak time of dyestuff and then controlling the painted depth, finally reaches target effect.But, adopt the method for soaking and dyeing to carry out dyeing to section at present and there is following problem:
1) by soaking, dye molecule free diffusing, the effect of diffusion alters a great deal, and the time can only control roughly;
2) time efficiency of soaking and dyeing is low, during operating cost;
3) poor contrast of dyestuff free diffusing, the contrast between signal (specific stain) and background noise (unspecific staining) is low, and identification is bad, easily causes false-positive result;
4) consumption of dyestuff is large, and cost is very high.
Summary of the invention
An object of the present invention is for above-mentioned deficiency, provides a kind of section statining method, and the method to employing soaking and dyeing to be solved is long to dyeing time during section statining, efficiency is low, and the problem that dye dosage is large, cost is high.Meanwhile, the present invention also provides a kind of baffle structure for section statining.
The object of invention is achieved through the following technical solutions:
A kind of baffle structure for section statining, comprise baffle main body, described baffle main body is provided with advection groove, and described advection groove one end is provided with the feed liquor dashpot be connected with advection groove, and the other end of this advection groove is provided with the fluid dashpot be connected with this advection groove; The sidewall of described feed liquor dashpot is provided with inlet opening, and the sidewall of described fluid dashpot is provided with fluid hole; The degree of depth of described advection groove is 150-250 microns.
According to one embodiment of present invention, it is characterized in that described feed liquor dashpot and described fluid dashpot symmetrically, and described inlet opening is identical with fluid hole size.
According to one embodiment of present invention, be connected with the advection groove width of one end of described feed liquor dashpot is greater than the width of the feed liquor dashpot other end, and described inlet opening is arranged on the other end of feed liquor dashpot.
According to one embodiment of present invention, the degree of depth of described advection groove is 180-220 microns.
According to one embodiment of present invention, the degree of depth of described advection groove is 200 microns.
According to one embodiment of present invention, described baffle body comprises metal material or glass are made.
A kind of section statining method, comprises the following steps:
(1) biological sample is cut into slices, and by sample slice sticker on microslide;
(2) microslide is placed in baffle main body, sample is cut into slices downwards and sample is cut into slices the advection groove be arranged on baffle;
(3) in diversion trench, import dyestuff by the inlet opening on baffle, dyestuff submergence sample is cut into slices;
(4) continue import dyestuff, make dyestuff at the uniform velocity advection cut into slices through sample;
(5) lower a kind of dyestuff is changed after completing the dyeing time of dyestuff, and circulation step (3)-(5), until complete the staining procedure of all dyestuffs.
According to one embodiment of present invention, in step (4) dyestuff through the flow velocity that sample is cut into slices be 3-8 cels.
According to one embodiment of present invention, in step (4) dyestuff through the flow velocity that sample is cut into slices be 5 cels.
According to one embodiment of present invention, adopt the dyeing time of Automatic control of single chip microcomputer dyestuff in step (5) and automatically control to change dyestuff.
The present invention comparatively prior art compares, and has the following advantages and beneficial effect:
(1) the present invention is used for the baffle of section statining not only structure is simple, and it is with low cost, when using baffle of the present invention to dye to sample section, microslide is placed in baffle main body, sample section is arranged in advection groove, dyestuff in advection groove then can form the reaction cabin of 200 microns thickness to biological dye, continuing in advection groove, import dyestuff then can make dyestuff form the flow field of stable advection, dyestuff is at the uniform velocity cut into slices through sample section and to sample and is dyeed, baffle structure of the present invention is used to dye to sample section, the contrast of dyestuff free diffusing is high, contrast between signal and background noise is high, identification is good, reduce false positive rate, the rate of propagation of dyestuff in flow field is faster than rate of propagation during static immersing, and the sample section statining time is short, can improve staining efficiency, also can save dye dosage, meanwhile, the flow velocity that dyestuff is cut into slices through sample can accurately control, and therefore also can improve the degree of accuracy of dyeing.
(2) feed liquor dashpot of the present invention and described fluid dashpot symmetrical, and inlet opening is identical with fluid hole size, relative buffering can be carried out with during derivation advection groove to dyestuff being imported by dyestuff advection groove, to ensure that dyestuff is formed at the uniform velocity and the flow field of advection in advection groove, the degree of accuracy to sample section statining can be ensured.
(3) be connected with the advection groove width of one end of described feed liquor dashpot of the present invention is greater than the width of the feed liquor dashpot other end, and described inlet opening is arranged on the other end of feed liquor dashpot; Can cushion fully in feed liquor dashpot after dyestuff can be made to enter feed liquor dashpot by inlet opening, to ensure that dyestuff is already formed at the uniform velocity when entering advection groove and the flow field of advection.
(4) degree of depth of advection groove of the present invention is 180-220 microns, can ensure that dyestuff can be cut into slices with sample in advection groove to react fully, namely sample section is dyeed, dyestuff can also be made in advection groove to form stable flow field, so that control accurately dyeing simultaneously.
(5) baffle body comprises metal material of the present invention or glass are made, can conveniently process on the one hand, can fit fully with baffle main body when microslide can be made to be placed in baffle main body on the other hand, the sample on microslide can be made to cut into slices and to be arranged in advection groove completely, to ensure that dyestuff can flood sample section completely, dyestuff can be cut into slices with sample and react fully.
(6) colouring method step of the present invention is simple, convenient operation, accurately can control dyeing, can also reduce false positive rate, also can save dye dosage simultaneously, and can improve the degree of accuracy of dyeing.
(7) in step of the present invention (4) dyestuff through the flow velocity that sample is cut into slices be 3-8 cels, the rate of propagation of dyestuff in flow field is faster than rate of propagation during static immersing, therefore can ensure that dyestuff and sample are cut into slices fully to react and can also improve staining efficiency, be convenient to accurately control dyeing.
(8) adopt the dyeing time of Automatic control of single chip microcomputer dyestuff in step of the present invention (5) and automatically control to change dyestuff, adopting the intelligent equipment automatically controlled to carry out dying operation, can cost of labor be saved, the degree of accuracy dyeed can also be ensured.
Accompanying drawing explanation
Fig. 1 is one-piece construction schematic diagram of the present invention.
Fig. 2 is the vertical view of Fig. 1.
Fig. 3 is the vertical view of one embodiment of the present invention.
Fig. 4 is the vertical view of one embodiment of the present invention.
Fig. 5 is the vertical view of one embodiment of the present invention.
Wherein, the name corresponding to the Reference numeral in accompanying drawing is called:
1-baffle main body, 2-advection groove, 3-feed liquor dashpot, 4-inlet opening, 5-fluid dashpot, 6-fluid hole.
Embodiment
Below in conjunction with embodiment, the present invention is described in further detail:
Embodiment
As shown in Figure 1, 2, a kind of baffle structure for section statining of the present invention, comprise baffle main body 1, described baffle main body 1 is made by metal material or glass.Can conveniently process on the one hand, can fit fully with baffle main body when microslide can be made to be placed in baffle main body on the other hand, the biologic slice on microslide can be made to be arranged in advection groove completely, to ensure that dyestuff can flood biologic slice completely, dyestuff can be reacted fully with biologic slice.For the ease of fitting with microslide, the upper surface of described baffle main body 1 is surface level.
Described baffle main body 1 is provided with advection groove 2, and described advection groove 2 is overall rectangular.In order to ensure that dyestuff forms stable flow field in advection groove, the degree of depth of described advection groove 2 is set to 150-250 microns, the degree of depth of advection groove is set to 150-250 microns, can ensure that dyestuff can react fully with biologic slice in advection groove 2, namely biologic slice is dyeed, dyestuff can also be made in advection groove to form stable flow field, so that control accurately dyeing simultaneously.More excellent selection is that the degree of depth of described advection groove 2 is set to 180-220 microns, and the degree of depth of the advection groove 2 in the present embodiment is 200 microns.
For the ease of cushioning dyestuff when being imported in advection groove 2 by dyestuff, one end of described advection groove 2 is provided with the feed liquor dashpot 3 be connected with advection groove 2, and the other end of this advection groove 2 is provided with the fluid dashpot 5 be connected with this advection groove 2 simultaneously.The sidewall of described feed liquor dashpot 3 is provided with inlet opening 4, and the sidewall of described fluid dashpot 5 is provided with fluid hole 6.Described inlet opening 4 is identical with the size of fluid hole 6, and inlet opening 4 of the present invention is circular port with fluid hole 6, and namely described inlet opening 4 is identical with the diameter of fluid hole 6.
Continue dyestuff to import in feed liquor dashpot 3 by inlet opening 4 during use, dyestuff is derived after the buffering by fluid dashpot 5 through fluid hole 6 again.In order to ensure the flowing that dyestuff is at the uniform velocity stable in advection groove 2, described feed liquor dashpot 3 is symmetrical with described fluid dashpot 5.Concrete, be connected with advection groove 2 width of one end of described feed liquor dashpot 3 is greater than the width of feed liquor dashpot 3 other end, and described inlet opening 4 is arranged on the other end of feed liquor dashpot 3.Inlet opening 4 is arranged on the other end of feed liquor dashpot 3, the displacement of flowing after dyestuff can be made to enter feed liquor dashpot 3 by inlet opening 4 is maximum, dyestuff can be made to cushion fully in feed liquor dashpot 3, to ensure that dyestuff is already formed at the uniform velocity when entering advection groove and the flow field of advection.Be connected with advection groove 2 width of the width of one end and advection groove 2 of feed liquor dashpot 3 of the present invention is identical, and described feed liquor dashpot 3 entirety can in trapezoidal, as shown in Figure 2; Described feed liquor dashpot 3 entirety also can be triangular in shape, as shown in Figure 3; Described feed liquor dashpot 3 be connected with advection groove 2 one end and this feed liquor dashpot 3 the other end between can also arrange curved, as shown in fig. 4 or 5.Same, be connected with advection groove 2 width of one end of described fluid dashpot 5 is greater than the width of fluid dashpot 5 other end, and the set-up mode of described fluid hole 6 is identical with the set-up mode of inlet opening 4, and this fluid hole 6 is arranged on the other end of fluid dashpot 5.
Be placed in baffle main body 1 by microslide when using baffle of the present invention to dye to biologic slice, biologic slice is arranged in advection groove 2, and the dyestuff in advection groove 2 then can form to biological dye the reaction cabin that thickness is 200 microns.Continue to import dyestuff by inlet opening 4, can ensure that under feed liquor dashpot 3 buffer action with fluid dashpot 5 the at the uniform velocity stable advection of dyestuff is through biologic slice, can make dyestuff form the flow field of stable advection, dyestuff at the uniform velocity dyes to biologic slice through biologic slice.Use baffle structure of the present invention to dye to biologic slice, the contrast of dyestuff free diffusing is high, and the contrast between signal and background noise is high, and identification is good, reduces false positive rate; The rate of propagation of dyestuff in flow field is faster than rate of propagation during static immersing, and biologic slice dyeing time is short, can improve staining efficiency, also can save dye dosage; Meanwhile, dyestuff can accurately control through the flow velocity of biologic slice, therefore also can improve the degree of accuracy of dyeing.
Section statining method of the present invention, first cuts into slices biological sample, and by sample slice sticker on microslide, sample section is inverted and also can not be dropped.Then microslide is placed in baffle main body 1, sample is cut into slices downwards and sample cut into slices the advection groove 2 be arranged on baffle, preferably making sample section be positioned at the centre position of advection groove 2.In diversion trench 2, import dyestuff by the inlet opening 4 on baffle, dyestuff submergence sample is cut into slices.Then continue to import dyestuff, make dyestuff at the uniform velocity advection cut into slices through sample, the flow control that dyestuff is cut into slices through sample by the present invention is at 3-8 cels.The rate of propagation of dyestuff in flow field is faster than rate of propagation during static immersing, at 3-8 cels, dyestuff can be ensured that dyestuff and sample are cut into slices through the flow control that sample is cut into slices fully react and can also improve staining efficiency, is convenient to accurately control dyeing.In the present embodiment dyestuff through the flow velocity that sample is cut into slices be 5 cels.
A kind of dyestuff under changing after completing the dyeing time of dyestuff, and repeat above-mentioned staining procedure, the step in advection groove, sample dyeed is imported by dyestuff.Change lower a kind of dyestuff after completing dyeing time to dye, continue to repeat above-mentioned staining procedure, until complete the staining procedure of all dyestuffs.The present invention adopts Automatic control of single chip microcomputer dyeing time and automatically controls to change dyestuff, input pipe dyestuff being inputted baffle inside of the present invention is set during use on inlet opening, input pipe connects many conduits respectively, a kind of dyestuff carried by each root conduit, and every root conduit is all provided with solenoid valve.Use the dyeing time of Single-chip Controlling dyestuff, after dyestuff completes dyeing time, Single-chip Controlling carries the supravasal closed electromagnetic valve of this kind of dyestuff, and the supravasal solenoid valve controlling the lower a kind of dyestuff of conveying is opened simultaneously, namely automatically replaceable lower a kind of dyestuff dyes to sample section.Use Automatic control of single chip microcomputer dyeing time and automatically control to change dyestuff, can cost of labor be saved, the degree of accuracy dyeed can also be ensured.
The foregoing is only preferred embodiment of the present invention, not in order to limit the present invention, all any amendments done within the spirit and principles in the present invention, equivalent replacement and improvement etc., all should be included within protection scope of the present invention.
Claims (10)
1. the baffle structure for section statining, it is characterized in that comprising baffle main body (1), described baffle main body (1) is provided with advection groove (2), described advection groove (2) one end is provided with the feed liquor dashpot (3) be connected with advection groove (2), and the other end of this advection groove (2) is provided with the fluid dashpot (5) be connected with this advection groove (2); The sidewall of described feed liquor dashpot (3) is provided with inlet opening (4), and the sidewall of described fluid dashpot (5) is provided with fluid hole (6); The degree of depth of described advection groove (2) is 150-250 microns.
2. a kind of baffle structure for section statining according to claim 1, it is characterized in that described feed liquor dashpot (3) is symmetrical with described fluid dashpot (5), and described inlet opening (4) is identical with fluid hole (6) size.
3. a kind of baffle structure for section statining according to claim 2, it is characterized in that be connected with advection groove (2) width of one end of described feed liquor dashpot (3) is greater than the width of feed liquor dashpot (3) other end, described inlet opening (4) is arranged on the other end of feed liquor dashpot (3).
4. a kind of baffle structure for section statining according to any one of claim 1-3, is characterized in that the degree of depth of described advection groove (2) is 180-220 microns.
5. a kind of baffle structure for section statining according to claim 4, is characterized in that the degree of depth of described advection groove (2) is 200 microns.
6. a kind of baffle structure for section statining according to claim 5, is characterized in that described baffle main body (1) is made by metal material or glass.
7. a section statining method, is characterized in that comprising the following steps:
(1) biological sample is cut into slices, and by sample slice sticker on microslide;
(2) microslide is placed in baffle main body, sample is cut into slices downwards and sample is cut into slices the advection groove be arranged on baffle;
(3) in diversion trench, import dyestuff by the inlet opening on baffle, dyestuff submergence sample is cut into slices;
(4) continue import dyestuff, make dyestuff at the uniform velocity advection cut into slices through sample;
(5) lower a kind of dyestuff is changed after completing the dyeing time of dyestuff, and circulation step (3)-(5), until complete the staining procedure of all dyestuffs.
8. a kind of section statining method according to claim 7, to it is characterized in that in step (4) dyestuff through the flow velocity that sample is cut into slices be 3-8 cels.
9. a kind of section statining method according to claim 8, to it is characterized in that in step (4) dyestuff through the flow velocity that sample is cut into slices be 5 cels.
10. a kind of section statining method according to any one of claim 7-9, is characterized in that adopting the dyeing time of Automatic control of single chip microcomputer dyestuff in step (5) and automatically controls to change dyestuff.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510981522.9A CN105424449A (en) | 2015-11-11 | 2015-12-23 | Flow-guiding body structure used for section staining and section staining method |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2015107653319 | 2015-11-11 | ||
| CN201510765331 | 2015-11-11 | ||
| CN201510981522.9A CN105424449A (en) | 2015-11-11 | 2015-12-23 | Flow-guiding body structure used for section staining and section staining method |
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| CN105424449A true CN105424449A (en) | 2016-03-23 |
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN2148957Y (en) * | 1992-10-08 | 1993-12-08 | 张敬雄 | Tank type stainer |
| EP1260265A1 (en) * | 2001-05-25 | 2002-11-27 | Tecan Trading AG | Apparatus for the preparation of a hybridisation chamber, processing set and system for the hybridisation of samples of nucleic acid, proteines and tissues |
| CN203616184U (en) * | 2013-12-10 | 2014-05-28 | 胡勤星 | Continuous leaching detection preprocessing device for fertilizer |
| CN104007238A (en) * | 2014-06-10 | 2014-08-27 | 北京航空航天大学 | Film surface uniform shearing force loading device |
| CN104136123A (en) * | 2012-01-09 | 2014-11-05 | 精密公司 | Microfluidic reactor system |
| CN205333384U (en) * | 2015-11-11 | 2016-06-22 | 刘洋 | A baffle structure for section statining |
-
2015
- 2015-12-23 CN CN201510981522.9A patent/CN105424449A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN2148957Y (en) * | 1992-10-08 | 1993-12-08 | 张敬雄 | Tank type stainer |
| EP1260265A1 (en) * | 2001-05-25 | 2002-11-27 | Tecan Trading AG | Apparatus for the preparation of a hybridisation chamber, processing set and system for the hybridisation of samples of nucleic acid, proteines and tissues |
| CN104136123A (en) * | 2012-01-09 | 2014-11-05 | 精密公司 | Microfluidic reactor system |
| CN203616184U (en) * | 2013-12-10 | 2014-05-28 | 胡勤星 | Continuous leaching detection preprocessing device for fertilizer |
| CN104007238A (en) * | 2014-06-10 | 2014-08-27 | 北京航空航天大学 | Film surface uniform shearing force loading device |
| CN205333384U (en) * | 2015-11-11 | 2016-06-22 | 刘洋 | A baffle structure for section statining |
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Application publication date: 20160323 |