CN101876748A - Reusable special cell culture dish of laser confocal microscope - Google Patents
Reusable special cell culture dish of laser confocal microscope Download PDFInfo
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- CN101876748A CN101876748A CN200910082988XA CN200910082988A CN101876748A CN 101876748 A CN101876748 A CN 101876748A CN 200910082988X A CN200910082988X A CN 200910082988XA CN 200910082988 A CN200910082988 A CN 200910082988A CN 101876748 A CN101876748 A CN 101876748A
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Abstract
The invention belongs to a cell culture dish, in particular relating to a reusable special cell culture dish of a laser confocal microscope. The cell culture dish is provided with a lower gasket at the bottom of a concave cell culture dish, the bottom of the concave cell culture dish is provided with a light through hole, a piece of cover glass is arranged on the lower gasket, an upper gasket is arranged on the cover glass, a cell culture dish core is arranged on the lower gasket, and the cell culture dish core is combined with the bottom of the concave cell culture dish in a sealing manner; the upper gasket, the lower gasket and the cell culture dish core are provided with light through holes same with the light though hole at the bottom of the cell culture dish in size but opposite in position; and a cell culture dish cover covers the cell culture dish core. In the invention, the main body material of the culture dish is stainless steel which does not react with the common cell culture solution and is easy to clean, thereby realizing the reusing of the culture dish main body; and the function of a porous culture plate can be realized by a porous structure.
Description
Technical field
The invention belongs to the cellular incubation vessel, particularly a kind of reusable special cell culture dish of laser confocal microscope.
Background technology
Cell is the basic structure and the functional unit of all vital movements.And microscope is the main tool of observation of cell, also is the main method of carrying out life science.Laser confocal microscope is widely used in the life science as a kind of modern detecting in recent years.The operating distance of the object lens of laser confocal microscope commonly used is all smaller now, particularly enlargement factor is at the object lens more than 60 times, operating distance is all below 0.18mm, and mostly common disposable Tissue Culture Dish is to adopt the transparent polymer material polystyrene material, and light transmission is poor, double dish wall thicker (about about 1mm) is not suitable for observing and imaging with laser confocal microscope.The burnt special-purpose double dish of the copolymerization of selling in the market all is that to adopt thickness be double dish at the bottom of the thick glass of 0.13～0.17mm, this double dish structure is to make a call to the hole that a diameter is 10mm in common disposable Tissue Culture Dish bottom, be stained with the square cover glass that a slice thickness is 0.13～0.17mm with xanthan gum then at the bottom of double dish, this double dish can only disposablely use.Also do not have big breakthrough owing to the technology of xanthan gum is domestic, this double dish of present domestic use mainly depends on import, and brings a high price, about 36 yuan/.This all is the no small expense of pen for the entity and individual that are engaged in life science and correlative study thereof.Up to now also not occurring commercialization both at home and abroad can reusable special cell culture dish of laser confocal microscope.
Thereby develop reusable special cell culture dish of laser confocal microscope and will provide a kind of economical and practical cellular incubation instrument for the research that all living creatures order science.
Summary of the invention
The objective of the invention is to overcome the defective of prior art, the employing stainless steel material is a main body, the main body of Tissue Culture Dish is divided into three parts, utilize precise machine machining to realize the combination of three parts and cover glass, thereby a kind of multi-functional reusable special cell culture dish of laser confocal microscope is provided.
Reusable special cell culture dish of laser confocal microscope of the present invention, comprise Tissue Culture Dish lid, matrix Tissue Culture Dish at the bottom of, the Tissue Culture Dish core, go up lower gasket, cover glass.
Be placed with described lower gasket at the bottom of the bottom has the described matrix Tissue Culture Dish of light hole, on lower gasket, be placed with described cover glass, on cover glass, be placed with described Upper gasket, on Upper gasket, be placed with described Tissue Culture Dish core, and the Tissue Culture Dish core combines with sealing at the bottom of the matrix Tissue Culture Dish; Have on described Upper gasket, lower gasket and the Tissue Culture Dish core with at the bottom of the matrix Tissue Culture Dish on the identical and corresponding light hole in position of light hole size; At Tissue Culture Dish core loam cake described Tissue Culture Dish lid is arranged.
The outward flange of described Tissue Culture Dish core has external thread, and the inboard of the matrix wall at the bottom of the matrix Tissue Culture Dish has internal thread, the internal thread engages and seals of the matrix wall at the bottom of the external thread of Tissue Culture Dish core and the matrix Tissue Culture Dish.
At the bottom of described Tissue Culture Dish lid, Tissue Culture Dish core and the matrix Tissue Culture Dish all is to form through finishing with stainless steel material.
Described Upper gasket and lower gasket all are to be processed by polytetrafluoroethylmaterial material.
Described cover glass is a laser confocal microscope special cap slide, and thickness can be 0.13～0.17mm.Detected cultured cell is carried on (one side that contacts with Upper gasket) on the cover glass.
The size of described cover glass is big or small the same or smaller with Upper gasket or lower gasket.
Above-mentioned light hole can be that circular port also can be a square opening, and the size in hole and quantity can design according to actual needs.As:
Can be designed to diameter at the bottom of the described matrix Tissue Culture Dish as required is 25mm, 30mm, 35mm, 40mm, 45mm, 50mm etc.; The diameter of light hole can be 10mm, 15mm, 20mm, 25mm, 30mm, 35mm, 40mm, 45mm etc.; The quantity of light hole can be 1～8 etc.
It is 24.8mm, 29.8mm, 34.8mm, 39.8mm, 44.8mm, 49.8mm etc. that described Tissue Culture Dish core can be designed to diameter as required; The diameter of light hole can be 10mm, 15mm, 20mm, 25mm, 30mm, 35mm, 40mm, 45mm etc.; The quantity of light hole can be 1～8 etc.
It is 24.8mm, 29.8mm, 34.8mm, 39.8mm, 44.8mm, 49.8mm etc. that described upward lower gasket can be designed to diameter as required, the diameter of light hole can be 10mm, 15mm, 20mm, 25mm, 30mm, 35mm, 40mm, 45mm etc., and the quantity of light hole can be 1～8 etc.
It is 26mm, 31mm, 36mm, 41mm, 46mm, 51mm etc. that described Tissue Culture Dish lid can be designed to diameter as required.
The present invention is suitable for adopting laser confocal microscope to detect cellular incubation, can realize the function of porous plate.
The present invention has:
(1) the present invention adopts the material of main part of stainless steel material as double dish, and any reaction does not take place for this material and conventional cell culture fluid, and cleans easily, has realized the repeated use of double dish main body;
(2) adopt polytetrafluoroethylmaterial material to do lower gasket, and guaranteed the body seal of double dish at the bottom of Tissue Culture Dish core and the Tissue Culture Dish by screw-threaded engagement;
(3) porous structure of the present invention can be realized the function of porous culture plate;
(4) apparatus structure of the present invention is simple, and easy operating is economical and practical, can be applied to a plurality of fields such as chemistry, biology, medical science, material, environment, safety.For the further investigation of the problem in science of nano science, material science, biological chemistry and crossing domain provides the high-level efficiency research means.
Description of drawings
Fig. 1. reusable special cell culture dish of laser confocal microscope structural representation of the present invention.
Fig. 2 (a). the attached cell with the Tissue Culture Dish cultivation of the embodiment of the invention 1 is observed the image of its light field under laser confocal microscope.
Fig. 2 (b). the attached cell of cultivating with Tissue Culture Dish of the embodiment of the invention 1 after adding fluorescence probe HgL, the fluorogram of under laser confocal microscope, observing.
Reference numeral
1. Tissue Culture Dish covers 4. cover glasses at the bottom of 2. light holes, 3. Tissue Culture Dishs
5. Tissue Culture Dish core 6. Upper gaskets 7. lower gaskets
Embodiment
The present invention will be further explained and explanation below in conjunction with specific embodiment.
Embodiment 1
See also Fig. 1.Reusable special cell culture dish of laser confocal microscope comprises at the bottom of Tissue Culture Dish lid 1, the matrix Tissue Culture Dish 3, Tissue Culture Dish core 5, Upper gasket 6, lower gasket 7, cover glass 4.
At the bottom of having the matrix Tissue Culture Dish that is prepared into by stainless steel material of light hole 2, the bottom is placed with the lower gasket 7 that processes by polytetrafluoroethylmaterial material in 3, on lower gasket, be placed with laser confocal microscope special cap slide 4, the thickness of cover glass is 0.13～0.17mm, on cover glass, be placed with the Upper gasket 6 that processes by polytetrafluoroethylmaterial material, (its effect is with Upper gasket 6 to be placed with the Tissue Culture Dish core 5 that is prepared into by stainless steel material on Upper gasket, lower gasket 7 and cover glass 4 compress), and the outward flange of Tissue Culture Dish core has external thread, the inboard of the matrix wall at the bottom of the matrix Tissue Culture Dish has internal thread, the internal thread engages and seals of the matrix wall inboard at the bottom of the external thread of Tissue Culture Dish core and the matrix Tissue Culture Dish; Have on described Upper gasket, lower gasket and the Tissue Culture Dish core with at the bottom of the Tissue Culture Dish on the identical and corresponding circular light hole in position of light hole size; On the Tissue Culture Dish core, be stamped the Tissue Culture Dish lid 1 that is prepared into by stainless steel material.
Adopt this Tissue Culture Dish, can in CO2gas incubator, cultivate attached cell and attached cell not.
Fig. 2 (a) and Fig. 2 (b) are the laser co-focusing picture of the HEK293 cell of the reusable special cell culture dish of laser confocal microscope cultivation of employing.The HEK293 cell that purchase obtains, is cultivated 37 ℃ of temperature, CO as nutrient culture media with MEM nutrient solution (mass concentration is 10%PBS, 2mM L-glu) in the torticollis Tissue Culture Flask
2Mass concentration be 5%, changed liquid once every 3 days.Go down to posterity, frozen standby.The test proxima luce (prox. luc), the frozen HEK293 cell of recovering, choose the good HEK293 cell of growth conditions, play dispersion with the trypsinization after-blow, be contained in (one side that contacts with Upper gasket) back encapsulated cell double dish on the cover glass in the Tissue Culture Dish behind disinfection by ultraviolet light with said structure then, wherein, diameter at the bottom of the matrix Tissue Culture Dish is 40mm, the thickness of cover glass is 0.13mm, the quantity of light hole is 1, the diameter of light hole is 30mm, and the diameter of Tissue Culture Dish lid is 41mm; The above-mentioned Tissue Culture Dish that the HEK293 cell is housed was cultivated in CO2gas incubator 24 hours, observed under ordinary optical microscope, most of cell is adherent state; With PBS damping fluid (phosphate buffer) nutrient culture media is rinsed out before the experiment, be placed on then and observe its light field image under the Laser Scanning Confocal Microscope shown in Fig. 2 (a), add the biological sulfhydryl compound fluorescence probe HgL of identification (referring to CN200810112996.X, " detect the fluorescence probe and the preparation method and use thereof of glutathione reductase and bioactive sulfhydryl compound ") hatched 5 minutes, obtain laser co-focusing fluorogram 2 (b), from Fig. 2 (a) and two figure of Fig. 2 (b), can find out that cell growth state is good, can under laser confocal microscope, be washed to picture.
Employing has the reusable special cell culture dish of laser confocal microscope of embodiment 1 structure, and wherein cover glass adopts the in situ hybridization cover glass, can realize the hybridization in situ experiment observation.
Employing has the reusable special cell culture dish of laser confocal microscope of embodiment 1 structure, and wherein cover glass adopts microslide, can cultivate various kinds of cell and be used for common biological microscopic examination and imaging.
Embodiment 4
Employing has the reusable special cell culture dish of laser confocal microscope of embodiment 1 structure, and wherein cover glass adopts the quartz cover slide, can cultivate various kinds of cell and be used for ultraviolet Laser Scanning Confocal Microscope and general ultraviolet fluorescence microscope and imaging.
Embodiment 5
Employing has the reusable special cell culture dish of laser confocal microscope of embodiment 1 structure, and wherein cover glass adopts silication cap fall prevention slide, and the fluorescence labeling co-focusing imaging that can be used in antigen retrieval process process is observed.
Embodiment 6
The cover glass that attached cell is arranged that will have in the reusable special cell culture dish of laser confocal microscope of embodiment 1 structure takes out, can be directly with the fixing mounting of attached cell; And a plurality of light holes in the reusable special cell culture dish of laser confocal microscope can be different test environments, can obtain the many mountings sample under the different test environments.
Embodiment 7
Employing has the reusable special cell culture dish of laser confocal microscope of embodiment 1 structure, and the outer wall at the bottom of Tissue Culture Dish sticks Peltier, reusable special cell culture dish of laser confocal microscope can be embodied as the constant temperature culture ware by circuit design.
Embodiment 8
The reusable special cell culture dish of laser confocal microscope that will have embodiment 1 structure is designed to rectangle, and have more light hole (6 holes, 12 holes, 24 holes, 48 holes, 72 holes, 96 holes) by requirement of experiment, can realize the function of many culture plates in 6 holes, 12 holes, 24 holes, 48 holes, 72 holes, 96 holes.
Embodiment 9
Employing has the reusable special cell culture dish of laser confocal microscope of embodiment 1 structure, and wherein cover glass adopts poly-D-lysine sheet base, is suitable for the imaging of widow's nucleotide probe fluorescence co-focusing.
Embodiment 10
Employing has the reusable special cell culture dish of laser confocal microscope of embodiment 1 structure, and wherein cover glass adopts the aldehyde radical glass chip, is suitable for amido modified dna probe co-focusing imaging.
Embodiment 11
Employing has the reusable special cell culture dish of laser confocal microscope of embodiment 1 structure, and wherein cover glass adopts isothiocyanic acid sheet or agar sugar-tablet base, polypeptide, micromolecule antigen co-focusing imaging in being suitable for.
Embodiment 12
Employing has the reusable special cell culture dish of laser confocal microscope of embodiment 1 structure, and wherein cover glass adopts the amination glass chip, is suitable for the imaging of dna probe fluorescence co-focusing.
Claims (7)
1. reusable special cell culture dish of laser confocal microscope is at the bottom of it comprises Tissue Culture Dish lid, matrix Tissue Culture Dish, the Tissue Culture Dish core, go up lower gasket, cover glass; It is characterized in that:
Be placed with described lower gasket at the bottom of the bottom has the described matrix Tissue Culture Dish of light hole, on lower gasket, be placed with described cover glass, on cover glass, be placed with described Upper gasket, on Upper gasket, be placed with described Tissue Culture Dish core, and the Tissue Culture Dish core combines with sealing at the bottom of the matrix Tissue Culture Dish; Have on described Upper gasket, lower gasket and the Tissue Culture Dish core with at the bottom of the matrix Tissue Culture Dish on the identical and corresponding light hole in position of light hole size; At Tissue Culture Dish core loam cake described Tissue Culture Dish lid is arranged.
2. reusable special cell culture dish of laser confocal microscope according to claim 1, it is characterized in that: the outward flange of described Tissue Culture Dish core has external thread, the inboard of the matrix wall at the bottom of the matrix Tissue Culture Dish has internal thread, the internal thread engages and seals of the matrix wall at the bottom of the external thread of Tissue Culture Dish core and the matrix Tissue Culture Dish.
3. according to claim 1 and 2 described reusable special cell culture dish of laser confocal microscope, it is characterized in that: all be to process at the bottom of described Tissue Culture Dish core and the matrix Tissue Culture Dish with stainless steel material.
4. reusable special cell culture dish of laser confocal microscope according to claim 1 is characterized in that: described Tissue Culture Dish lid is to process with stainless steel material.
5. reusable special cell culture dish of laser confocal microscope according to claim 1 is characterized in that: described Upper gasket and lower gasket all are to be processed by polytetrafluoroethylmaterial material.
6. reusable special cell culture dish of laser confocal microscope according to claim 1 is characterized in that: described cover glass is a laser confocal microscope special cap slide.
7. according to claim 1 or 6 described reusable special cell culture dish of laser confocal microscope, it is characterized in that: the thickness of described cover glass is 0.13～0.17mm.
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| Application Number | Priority Date | Filing Date | Title |
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| CN200910082988XA CN101876748A (en) | 2009-04-28 | 2009-04-28 | Reusable special cell culture dish of laser confocal microscope |
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| CN200910082988XA CN101876748A (en) | 2009-04-28 | 2009-04-28 | Reusable special cell culture dish of laser confocal microscope |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102226151A (en) * | 2011-05-13 | 2011-10-26 | 刘小龙 | Removable cell imaging device |
| CN102226150A (en) * | 2011-05-13 | 2011-10-26 | 刘小龙 | Cell cultivating apparatus used for imaging living cell |
| CN102925354A (en) * | 2012-11-06 | 2013-02-13 | 无锡耐思生物科技有限公司 | Slide culture dish used by slides in two specifications in combination mode |
| CN104830681A (en) * | 2014-02-09 | 2015-08-12 | 李保伟 | Cell culture glass slide system |
| CN110133837A (en) * | 2018-02-08 | 2019-08-16 | 中国科学院微生物研究所 | For microscopical load sample device and its application |
| CN111500421A (en) * | 2020-01-03 | 2020-08-07 | 上海钰森生物技术有限公司 | Nano phase change constant temperature culture dish and manufacturing method thereof |
| CN112501016A (en) * | 2020-11-02 | 2021-03-16 | 山东华谱检测技术有限公司 | Carrier liquid device of laser confocal microscope for culturing eukaryotic cells |
| CN116694441A (en) * | 2023-06-27 | 2023-09-05 | 广东省农业科学院植物保护研究所 | Insect cell culture dish convenient to take slide |
-
2009
- 2009-04-28 CN CN200910082988XA patent/CN101876748A/en active Pending
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102226151A (en) * | 2011-05-13 | 2011-10-26 | 刘小龙 | Removable cell imaging device |
| CN102226150A (en) * | 2011-05-13 | 2011-10-26 | 刘小龙 | Cell cultivating apparatus used for imaging living cell |
| CN102925354A (en) * | 2012-11-06 | 2013-02-13 | 无锡耐思生物科技有限公司 | Slide culture dish used by slides in two specifications in combination mode |
| CN104830681A (en) * | 2014-02-09 | 2015-08-12 | 李保伟 | Cell culture glass slide system |
| CN110133837A (en) * | 2018-02-08 | 2019-08-16 | 中国科学院微生物研究所 | For microscopical load sample device and its application |
| CN111500421A (en) * | 2020-01-03 | 2020-08-07 | 上海钰森生物技术有限公司 | Nano phase change constant temperature culture dish and manufacturing method thereof |
| CN112501016A (en) * | 2020-11-02 | 2021-03-16 | 山东华谱检测技术有限公司 | Carrier liquid device of laser confocal microscope for culturing eukaryotic cells |
| CN112501016B (en) * | 2020-11-02 | 2023-08-01 | 山东华谱检测技术有限公司 | Laser confocal microscope carrier liquid device for eukaryotic cell culture |
| CN116694441A (en) * | 2023-06-27 | 2023-09-05 | 广东省农业科学院植物保护研究所 | Insect cell culture dish convenient to take slide |
| CN116694441B (en) * | 2023-06-27 | 2024-03-08 | 广东省农业科学院植物保护研究所 | Insect cell culture dish convenient to take slide |
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Open date: 20101103 |