Cell experiment device

Abstract

The utility model discloses a cell experiment device which comprises a degassing water environment bearing structure, a culture dish supporting structure, a cell culture dish and an ultrasonic transducer, the culture dish supporting structure is used for bearing a cell culture dish and is arranged in the degassing water environment bearing structure, the water injection rate in the degassing water environment bearing structure at least submerges the cell culture dish, and the surface of the cell culture dish is covered with a plastic film so that the internal environment of the cell culture dish is isolated from the external environment; the ultrasonic transducer is arranged above the cell culture dish and is positioned on the same vertical plane with the cell culture dish. The cell experiment device can effectively meet the ultrasonic conduction rule and ensure that ultrasonic waves accurately act on cells; the pollution problem in the experiment process is effectively prevented, and the influence on the cell state is reduced; the device is simple and convenient to operate and good in universality, and can be matched with ultrasonic transducers with various center frequencies, focusing depths and apertures and interference of sound fields in various forms.

Description

Technical Field

[0001] This utility model relates to the fields of ultrasound therapy, sonodynamic therapy and ultrasound stimulation technology, and in particular to a cell experiment device in related research. Background Technology

[0002] Ultrasound is widely used in clinical practice due to its non-invasive nature and deep penetration. In recent years, the emergence of ultrasound intervention methods such as sonodynamic therapy (SDT) and ultrasound nerve stimulation has led to extensive research on ultrasound therapy and stimulation. Cell experiments, due to their high controllability and reproducibility, are often used in studies such as efficacy assessment, mechanism exploration, and parameter optimization. For the numerous cell experiments related to ultrasound therapy and stimulation, there is an urgent need for a stable, versatile, ultrasound conduction-compliant, highly reproducible, and easy-to-operate cell experiment device. Utility Model Content

[0003] This invention provides a stable, versatile, and highly reproducible cell experiment device that conforms to the laws of ultrasound conduction, is easy to operate, and is suitable for various ultrasound therapy and stimulation-related cell experiments.

[0004] To achieve the above objectives, this utility model proposes a cell experiment device, comprising a deaerated water environment support structure, a culture dish support structure, a cell culture dish, and an ultrasonic transducer; wherein,

[0005] The culture dish support structure carries the cell culture dish in the deaerated water environment support structure. The water volume in the deaerated water environment support structure is at least enough to cover the cell culture dish. The surface of the cell culture dish is covered with a plastic film to isolate the internal environment of the cell culture dish from the external environment.

[0006] The ultrasonic transducer is positioned above the cell culture dish and on the same vertical plane as the cell culture dish.

[0007] Optionally, the vertical distance between the ultrasonic transducer and the cell culture dish is adjustable.

[0008] Optionally, the plastic film is a PE film or a polyester film.

[0009] Optionally, the thickness of the plastic film is less than or equal to 10 μm.

[0010] Add culture medium to the cell culture dish, cover the surface of the cell culture dish with a PE film or polyester film with a thickness of 10 μm or less, and use a fixing device to fix the film to achieve the purpose of sealing the cell culture dish. The above device can effectively prevent the generation of air bubbles in the culture medium and prevent cell contamination, while also minimizing the impact of the cell experiment device on ultrasound conduction.

[0011] Optionally, the deaerated water environment support structure is an acrylic water tank.

[0012] Optionally, the water in the deaerated water environment support structure is bubble-free water.

[0013] Optionally, the culture dish support structure is a ring-shaped hollow support.

[0014] Optionally, the water environment depth in the deaerated water environment bearing structure is greater than 30cm.

[0015] This depth helps to minimize ultrasonic attenuation.

[0016] Optionally, the height of the ultrasonic transducer can be adjusted via a three-dimensional displacement platform.

[0017] This cell experiment device has a simple structure and can be used for various forms of ultrasound intervention experiments. It can ensure that cells are not contaminated by the external environment, greatly reducing the difficulty of ultrasound cell intervention experiments and improving their reproducibility.

[0018] This cell experiment device effectively satisfies the laws of ultrasound conduction when performing ultrasound intervention on cells, greatly reducing the impact on cell state. It features high repeatability, simple operation, and good versatility, and can be applied to ultrasound transducers of various frequencies, focusing depths, and apertures, as well as ultrasound intervention in various sound fields. Attached Figure Description

[0019] To more clearly illustrate the technical solutions in the embodiments of this utility model or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this utility model. For those skilled in the art, other drawings can be obtained based on the structures shown in these drawings without creative effort.

[0020] Figure 1(a) is a schematic diagram of the cell experiment apparatus;

[0021] Figure 1(b) is a schematic diagram of the process of covering the surface of a cell culture dish with a plastic film;

[0022] Figure 2 The diagram shows the energy change of the focused ultrasound field through a single-focus phased array with and without passing through a PE film.

[0023] Figure 3This is a statistical chart showing the apoptosis, necrosis, and apoptosis / necrosis ratio of RAW264.7-derived foam cells after low-intensity, phased-array focused ultrasound-mediated SDT followed by Hoechst 33342 / PI double staining using this cell experimental apparatus; * indicates P < 0.05 compared to the control group, **** indicates P < 0.0001 compared to the control group.

[0024] Explanation of icon numbers:

[0025] 1. Deaerated water environment support structure; 2. Culture dish support structure; 3. Cell culture dish; 4. Ultrasonic transducer;

[0026] The realization of the purpose, functional features and advantages of this utility model will be further explained in conjunction with the embodiments and with reference to the accompanying drawings. Detailed Implementation

[0027] The technical solutions of the present utility model will be clearly and completely described below with reference to the accompanying drawings of the embodiments. Obviously, the described embodiments are only some embodiments of the present utility model, and not all embodiments. Based on the embodiments of the present utility model, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the protection scope of the present utility model.

[0028] It should be noted that if the embodiments of this utility model involve directional indicators (such as up, down, left, right, front, back, etc.), the directional indicators are only used to explain the relative positional relationship and movement of the components in a certain specific posture (as shown in the figure). If the specific posture changes, the directional indicators will also change accordingly.

[0029] Furthermore, if the embodiments of this utility model involve descriptions such as "first" or "second," these descriptions are for descriptive purposes only and should not be construed as indicating or implying their relative importance or implicitly specifying the number of indicated technical features. Therefore, features defined with "first" or "second" may explicitly or implicitly include at least one of those features. Additionally, the technical solutions of the various embodiments can be combined with each other, but this must be based on the ability of those skilled in the art to implement them. If the combination of technical solutions is contradictory or impossible to implement, it should be considered that such a combination of technical solutions does not exist and is not within the scope of protection claimed by this utility model.

[0030] This invention proposes a cell experiment apparatus. Referring to Figures 1(a) and 1(b), it includes an acrylic water tank, a ring-shaped hollow support, a cell culture dish (3), and an ultrasonic transducer (4). The cell culture dish (3) is filled with culture medium, and the surface of the cell culture dish (3) is covered with a PE film with a thickness of less than or equal to 10 μm. Rubber bands are provided on the PE film to fix the PE film, thereby sealing the cell culture dish (3) and isolating its internal environment from the external environment. The ring-shaped hollow support carries the cell culture dish (3) in the acrylic water tank, and the water depth in the acrylic water tank is greater than or equal to 30 cm. The ultrasonic transducer (4) is located above the cell culture dish (3) and is on the same vertical plane as the cell culture dish (3). The height of the ultrasonic transducer (4) is adjusted by a three-dimensional displacement platform, thereby adjusting the vertical distance between the ultrasonic transducer (4) and the cell culture dish (3) to meet the intervention depth requirements of different ultrasonic transducers. In the ultrasound intervention experiment, since the cell culture dish (3) is covered with a PE film, the cell culture dish (3) can be replaced repeatedly, which effectively prevents the culture medium in the cell culture dish (3) from being contaminated during the experiment, and greatly increases the versatility and reproducibility of the cell experiment device.

[0031] Taking the low-intensity, phased-array focused ultrasound-mediated sonodynamic therapy experiment on RAW264.7-derived foam cells as an example, the application steps of this cell experimental device are as follows:

[0032] 1. Fill the acrylic water tank with bubble-free water and place a ring-shaped hollow support inside it;

[0033] 2. Place a metal sheet on the annular hollow support for ultrasonic transducer (4) leveling and calibration;

[0034] 3. Add DVDMS sonication agent to the end of cell culture dish (3) and incubate for 5 hours in serum-free DMEM high-glucose medium (0.8 μmol / L). Then discard the culture medium and add 5-6 mL of serum-free DMEM high-glucose medium to the 35 mm diameter cell culture dish (3). Carefully seal the culture dish with a 10 μm thick PE film. After removing air bubbles, tighten the PE film with a rubber sleeve.

[0035] 4. After the ultrasound intervention is completed, the cell culture dish (3) is taken out of the acrylic water tank, the PE film on the surface of the cell culture dish (3) is removed in the ultra-clean workbench, some culture medium is aspirated, and the dish is placed in the cell culture incubator for continued culture or for efficacy assessment.

[0036] Characterizing the shape of the single-focus and multi-focus phased array focused ultrasound sound field passing through and without the PE film, and characterizing the energy change of the single-focus phased array focused ultrasound sound field passing through and without the PE film, it was found that the PE film has no significant effect on the shape of the complex phased array focused sound field. Furthermore, although the influence of the PE film on the sound field energy gradually increases with increasing excitation voltage, the increase is small and linear. These experimental results indicate that covering the cell culture dish (3) with a PE film does not significantly affect the shape or energy of the ultrasound sound field acting on the cell surface.

[0037] Reference Figure 3 The control group received no treatment; the sonosensitive agent group received only sodium porphyrin 0.8 μmol / L for 2 hours; the ultrasound group received 0.8 W / cm² of sonosensitive agent. 2 The ultrasound radiation group received 5 minutes of ultrasound; the sonodynamic therapy group received 0.8 μmol / L sodium porphyrin for 4 hours of incubation, and 0.8 W / cm² of ultrasound. 2 The cells were subjected to ultrasound irradiation for 5 minutes. Six hours after ultrasound, Hoechst 33342 / PI staining was performed, and continuous in situ fluorescence imaging was conducted using a fluorescence microscope and a three-dimensional displacement platform to calculate the apoptosis and necrosis rates. The results showed that SDT using this cell experimental device significantly induced apoptosis in RAW264.7-derived foam cells, while no significant apoptosis was observed in the control group or the ultrasound-only group, demonstrating the effectiveness of this cell experimental device and its lack of impact on the cells themselves.

[0038] An experiment was conducted to intervene in RAW264.7-derived foam cells using SDT mediated by multi-point focused S-shaped sound field. Treatment conditions were 0.8 W / cm², intervention time 5 min, and Hoechst 33342 / PI staining 6 h later. Continuous in-situ fluorescence imaging was performed using a fluorescence microscope and a three-dimensional displacement platform. Characterization of the XY cross-section of the multi-focus phased array focused ultrasound sound field, and Hoechst 33342 / PI double staining after low-intensity, multi-focus phased array focused ultrasound-mediated SDT, revealed significant apoptotic regions in the cells. The results showed that this cell experimental device can ensure precise ultrasound targeting of cells, and the significant apoptotic regions corresponded to the contour of the phased array focused ultrasound sound field, further demonstrating the feasibility of this cell experimental device.

[0039] The above description is only a preferred embodiment of the present utility model and does not limit the patent scope of the present utility model. All equivalent structural transformations made under the inventive concept of the present utility model using the contents of the present utility model specification and drawings, or direct / indirect applications in other related technical fields, are included within the patent protection scope of the present utility model.

Claims

1. A cell experiment apparatus, characterized in that, It includes a deaerated water environment support structure (1), a culture dish support structure (2), a cell culture dish (3), and an ultrasonic transducer (4); among which, The culture dish support structure (2) supports the cell culture dish (3) and places it in the deaerated water environment support structure (1). The amount of water injected into the deaerated water environment support structure (1) is at least enough to cover the cell culture dish (3). The surface of the cell culture dish (3) is covered with a plastic film to isolate the internal environment of the cell culture dish (3) from the external environment. The ultrasonic transducer (4) is positioned above the cell culture dish (3) and is located on the same vertical plane as the cell culture dish (3).

2. The cell experiment apparatus according to claim 1, characterized in that, The vertical distance between the ultrasonic transducer (4) and the cell culture dish (3) is adjustable.

3. The cell experiment apparatus according to claim 1, characterized in that, The plastic film is a PE film or a polyester film.

4. The cell experiment apparatus according to claim 3, characterized in that, The thickness of the plastic film is less than or equal to 10 μm.

5. The cell experiment apparatus according to claim 1, characterized in that, The deaerated water environment support structure (1) is an acrylic water tank.

6. The cell experiment apparatus according to claim 1, characterized in that, The water in the deaerated water environment bearing structure (1) is bubble-free water.

7. The cell experiment apparatus according to claim 1, characterized in that, The water environment depth in the deaerated water environment bearing structure (1) is greater than 30cm.

8. The cell experiment apparatus according to claim 1, characterized in that, The culture dish support structure (2) is a ring-shaped hollow support.

9. The cell experiment apparatus according to claim 1, characterized in that, The height of the ultrasonic transducer (4) can be adjusted by a three-dimensional displacement platform.

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