CN102719385B - Novel application of cell culture device Opticell - Google Patents
Novel application of cell culture device Opticell Download PDFInfo
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- CN102719385B CN102719385B CN201210241266.6A CN201210241266A CN102719385B CN 102719385 B CN102719385 B CN 102719385B CN 201210241266 A CN201210241266 A CN 201210241266A CN 102719385 B CN102719385 B CN 102719385B
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- opticell
- support
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- bacterial biofilm
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
- C12M25/02—Membranes; Filters
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/06—Nozzles; Sprayers; Spargers; Diffusers
- C12M29/08—Air lift
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
- C12M35/04—Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli
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Abstract
The invention discloses an application of an Opticell device (2) in an ultrasonic microbubble interfere bacterial biofilm experiment device. The device comprises a vessel (1), a bacterial biofilm culture device, i.e., the Opticell device (2), a support (5), a support (6) and an ultrasonic therapy head (7), wherein the Opticell device (2) contains a microbubble layer (4) and a bacterial biofilm layer (3); the vessel (1) is internally provided with de-aerated water; the Opticell device (2) is inverted, and supported and fixed by using the support (5) and the support (6), and the Opticell device (2), the support (5) and the support (6) are all soaked in the de-aerated water; and the vessel (1) is internally provided with the ultrasonic therapy head (7). By using the Opticell device (2), the attenuation of ultrasonics is greatly reduced, ultrasonic energy can be effectively transferred to the microbubbles, the requirement that the microbubbles are in sufficient contact with bacterial biofilms is met, and the effectiveness of an experiment is improved.
Description
Technical field
The present invention relates to the new purposes of a kind of cell culture apparatus Opticell, relate in particular to it and intervene the application in bacterial biofilm experimental installation at ultrasonic microbubble.
Background technology
Bacterial biofilm (Biofilm, BF) is the corresponding growth pattern of a kind of and bacterioplankton that bacterium forms for adapting to living environment in process of growth, bacterium and the extracellular matrix of self secreting, consists of, and has 3-D solid structure.BF is considered to human body built-in medical equipment as the major reason of the infections relatings such as blood vessel built-in catheter, joint prosthesis.Large quantity research discovery, bacterium exists and can effectively escape human immune system and antibiotic removing with BF form, increases the resistance of bacterial population, and its resistance can reach 1000 times of bacterioplankton.This makes to infect chronic, persistence and the repetitious feature of presenting.
Classic flat-plate method culturing bacterium BF puts into sheet glass or plastic sheet (the about 1-2cm of area in the hole of culture dish
2) as the carrier of BF, then inoculate appropriate bacterial suspension, under suitable envrionment conditions, cultivate certain hour after bacterium can be attached on carrier and form the BF that area is suitable with carrier, the output that improve BF need increase hole or the number of times of inoculation.
Ultrasonic microbubble is the small spheroid that includes gas, and microvesicle vibration can occur under ul-trasonic irradiation or breaks, and consequent " acoustic horn effect " and " mechanical effect " are widely used in the research in the fields such as thrombolysis, tumour.The support that the safety and efficacy of ultrasonic microbubble technology in medical technology has been studied much.Adopt ultrasonic microbubble technology to suppress infected by microbes and belong to brand-new research field, relevant report seldom at present.If ultrasonic microbubble technology is fully confirmed acting in experiment in vitro of bacterium BF, will infect the new approach that provides for anti-bacteria BF.
The measure that the ultrasonic correlation technique of employing of existing bibliographical information is intervened BF is first with classic flat-plate culture method, in the bottom of culture dish hole or cell culture insert, to prepare bacterium BF, then suck nutrient solution, change to the suspension of medicine or medicine and microvesicle, ultrasound probe from culture dish bottom, upwards launches ultrasonic or to immerse liquid transmitting downwards ultrasonic ,+medicine ultrasonic to study and the impact of ultrasonic+microvesicle+medicine on BF.But existing method exists the shortcoming that ultrasonic attenuation degree is large and ultrasonic microbubble is excessive with BF operating distance, be there is to remarkably influenced in ultrasonic effect of intervening bacterium BF.
Opticell device is novel totally enclosed cell culture apparatus at present, for cell cultures.
Summary of the invention
The object of the present invention is to provide the application of Opticell device in bacterial biofilm is cultivated.
Another object of the present invention is to provide Opticell device to intervene the application in bacterial biofilm experimental installation at ultrasonic microbubble.
To achieve these goals, the present invention is by the following technical solutions:
The application of Opticell device in bacterial biofilm is cultivated.
Opticell device flies the cell culture apparatus that your science and technology (thermofisher) company of generation produces for match is silent, its volume is 10ml, and an area is respectively arranged at top and bottom is 50cm
2polystyrene film, at present all for the cultivation to cell; Contriver chances in research and development, obtain going out 50-100 BF doubly compared with classic flat-plate fado, and pollution rate is almost nil after 1 inoculated bacteria suspension under same incubation time.The bacterium BF quantity that adopts Opticell culture method to prepare is large, can extract enough DNA or RNA and carry out molecular biology research and need not carry out the amplification of bacterial number; Meanwhile, BF can also shear and be divided into distinct portions to meet the demand of several different experimental implementation simultaneously from Opticell.
Opticell device is intervened the application in bacterial biofilm experimental installation at ultrasonic microbubble, in described Opticell device, is injected with microscopic bubble layer, and bacterial biofilm layer is arranged at its bottom.
Opticell device is intervened the application in bacterial biofilm experimental installation at ultrasonic microbubble, and described Opticell device arranges for being inverted.
Above-mentioned Opticell device is intervened the application in bacterial biofilm experimental installation at ultrasonic microbubble, specifically, described ultrasonic microbubble is intervened bacterial biofilm culture apparatus Opticell, two stands and the ultrasonic therapeutic head that bacterial biofilm experimental installation comprises container, contains microscopic bubble layer and bacterial biofilm layer; Described internal tank has de aerated water; Described Opticell device, for being inverted, is fixed by described two stands support, and Opticell device, two stands are all immersed in de aerated water; In described container, be provided with described ultrasonic therapeutic head.
In order to simplify manufacture, described container can be designed to cubic.
In order to reach better ultrasonic transmission effect, described ultrasonic therapeutic head is arranged on the position at the corresponding Opticell device of container bottom center.
The present invention's beneficial effect specific as follows:
The present invention, by the application in bacterial biofilm is cultivated of Opticell device, has effectively improved the vitro culture efficiency of biofilm load.Opticell device of the present invention is intervened the application in bacterial biofilm experimental installation at ultrasonic microbubble, the structure of finding Opticell device uniqueness is conducive to carrying out of ultrasonic microbubble intervention bacterium BF experiment in vitro, Opticell device is totally-enclosed system, after injecting ultrasonic microbubble, thereby device is inverted and can be made microvesicle fully contact with BF, improved the validity of experiment; Polystyrene film on Opticell is to ultrasonic almost not decay, Opticell device is put into the container that de aerated water is housed, ultrasonic by upwards transmitting of container bottom, through de aerated water and Opticell membrane action in BF and microvesicle, ultrasonic energy decrement degree can significantly reduce, and the effect of ultrasonic intervention BF is improved greatly.Through comparison test: existing bibliographical information ultrasonic+microbiotic makes the bacterium alive in BF be reduced to 1/10, ultrasonic+microvesicle+microbiotic makes the viable bacteria in BF be reduced to 1/100; Opticell device is intervened in bacterial biofilm experimental installation and applied at ultrasonic microbubble, reduced the operating distance of ultrasonic decay and microvesicle and BF, under same antibiotic concentration, the effect of intervention is doubled, above numerical value becomes respectively 1/20 and 1/200.
Accompanying drawing explanation
Fig. 1 is that Opticell device is intervened in the application in bacterial biofilm experimental installation at ultrasonic microbubble, and ultrasonic microbubble is intervened the structural representation of bacterial biofilm experimental installation.
Embodiment
Below by embodiment, the present invention is specifically described; be necessary to be pointed out that at this following examples are only for being further detailed the present invention; can not be interpreted as limiting the scope of the invention, those skilled in the art can make some nonessential improvement and adjustment to the present invention according to foregoing.
Embodiment: Opticell device is intervened the application in bacterial biofilm experimental installation at ultrasonic microbubble, specifically, as shown in Figure 1, described ultrasonic microbubble is intervened bacterial biofilm experimental installation and is comprised container 1, bacterial biofilm culture apparatus Opticell device 2, two stands and ultrasonic therapeutic head 7; Described container 1 inside has de aerated water; Described Opticell device 2, for being inverted, is fixed by described two stands support, and Opticell device 2, two stands are all immersed in de aerated water; In described container 1, be provided with described ultrasonic therapeutic head 7.
In order to simplify manufacture, the container 1 in the present embodiment is cubic.
In order to reach better ultrasonic transmission effect, in the present embodiment, ultrasonic therapeutic head 7 is arranged on the position at corresponding Opticell device 2 centers, container 1 bottom.
As shown in Figure 1, two stands described in the present embodiment is support 5 and 6.
When experiment starts, first with Opticell device 2, set up bacterium BF external model.10ml is inoculated to bacterium liquid from Opticell device 2 side opening injecting chamber, Opticell device 2 is statically placed in to 37 ℃ of incubators, after cultivation appropriate time, can obtain bacterium BF.If with the Polystyrene Film of Opticell device 2 belows under scissors cut, available scanning electron microscope is identified the formation of bacterial biofilm.
After bacterium BF external model has been set up, microvesicle suspension is injected to inner chamber through Opticell device 2 side openings, be inverted Opticell device 2 microscopic bubble layer 4 floatings wherein are fully contacted with bacterial biofilm layer 3.Inverted Opticell device 2 is placed into the container 1 that inside has de aerated water, with support 5 and 6, supports to fix, be immersed in de aerated water.The position at corresponding Opticell device 2 centers, container 1 bottom is provided with ultrasonic therapeutic head 7.Bacterial biofilm 3 is fixed in hyperacoustic region by support 5 and support 6 like this, is beneficial to the enforcement of experiment.Polystyrene Film almost can see through ultrasonic wave completely, has greatly reduced hyperacoustic decay.
Claims (2)
1.Opticell
?device (2) is intervened the application in bacterial biofilm experimental installation at ultrasonic microbubble, and described experimental installation comprises container (1), contains bacterial biofilm culture apparatus Opticell device (2), support 5, support 6 and the ultrasonic therapeutic head (7) of microscopic bubble layer (4) and bacterial biofilm layer (3); Described container (1) inside has de aerated water; Described Opticell
?in device (2), be injected with microscopic bubble layer (4), bacterial biofilm layer (3) is arranged at its bottom; Described Opticell device (2), for being inverted, is supported to fix by described support 5 and support 6, and Opticell device (2), support 5 and support 6 are all immersed in de aerated water; In described container (1), be provided with described ultrasonic therapeutic head (7); Described ultrasonic therapeutic head (7) is arranged on the position at corresponding Opticell device (2) center, container (1) bottom.
2. application as claimed in claim 1, is characterized in that: described container (1) is designed to cubic.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210241266.6A CN102719385B (en) | 2012-07-12 | 2012-07-12 | Novel application of cell culture device Opticell |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210241266.6A CN102719385B (en) | 2012-07-12 | 2012-07-12 | Novel application of cell culture device Opticell |
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| Publication Number | Publication Date |
|---|---|
| CN102719385A CN102719385A (en) | 2012-10-10 |
| CN102719385B true CN102719385B (en) | 2014-01-15 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201210241266.6A Expired - Fee Related CN102719385B (en) | 2012-07-12 | 2012-07-12 | Novel application of cell culture device Opticell |
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Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106031795A (en) * | 2016-07-22 | 2016-10-19 | 浙江大学 | Ultrasound-microbubble transferring device and method thereof |
| WO2019113930A1 (en) * | 2017-12-15 | 2019-06-20 | 深圳先进技术研究院 | Ultrasonic microbubble cell stimulation device and operation method thereof |
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- 2012-07-12 CN CN201210241266.6A patent/CN102719385B/en not_active Expired - Fee Related
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