Confocal perforated plate for tube forming experiment

Abstract

The utility model discloses a confocal perforated plate for tube formation experiment, which comprises a base plate structure, accommodating hole structures and bulge structures, the accommodating hole structures are equidistantly arranged on the upper surface of the base plate structure, and the bulge structures are arranged at the bottoms of the inner walls of the accommodating hole structures. According to the confocal perforated plate for the tube forming experiment, self-leveling of matrigel is achieved through combination of micron-sized annular protrusion arrays designed at the bottoms of the holes and super-hydrophilic surface modification, the standard deviation of the thickness of a gel layer is controlled within + / -3 microns, and the height difference of the liquid level is reduced to lt through side wall gradient hydrophobic treatment (top hydrophobic / bottom hydrophilic); 0.1 mm, and halo artifacts in confocal imaging are eliminated.

Description

Technical Field

[0001] This utility model relates to the field of cell culture experimental equipment technology, specifically a confocal multiwell plate for tube formation experiments. Background Technology

[0002] In in vitro angiogenesis studies (i.e., "tube formation experiments"), endothelial cells need to be in a matrix gel (such as... A tubular structure is formed in a supported three-dimensional environment, and high-resolution dynamic imaging is achieved using a confocal microscope. Existing standard porous plates have the following technical drawbacks in this application:

[0003] 1. Imaging defects caused by difficulty in leveling the matrix gel.

[0004] Traditional multi-well plates have smooth, flat bottoms. Due to surface tension, liquid matrix gels tend to form a "coffee ring" distribution with thicker edges and a thinner center (thickness difference of 20-50 μm). Because confocal microscopes have extremely shallow depth of field, uneven gel layers can cause some areas of cell tubular structures to become out of focus, requiring multiple adjustments to the focus to achieve complete imaging, leading to errors in image stitching.

[0005] 2. Optical interference caused by sidewall capillary effect

[0006] The hydrophilicity (contact angle of approximately 75°) of the sidewall material of porous plates (such as polystyrene) causes liquid to climb along the wall surface due to capillary action, forming a centrally concave "meniscus" (with a liquid level difference of 0.5-1.2 mm). This liquid surface curvature alters the refractive index of the laser confocal system, causing light intensity attenuation in the central region of the image and producing a ring-shaped halo, which severely affects the quantitative analysis of fluorescence signals.

[0007] 3. Operational complexity and experimental stability issues

[0008] Abnormal edge shear forces caused by liquid surface distortion may induce a deflection of cell migration direction, affecting the reproducibility of tube formation experiments.

[0009] Therefore, we propose a confocal multi-well plate for tube forming experiments to solve the above problems. Utility Model Content

[0010] The purpose of this invention is to provide a confocal multi-well plate for tube forming experiments, so as to solve the problems mentioned in the background art.

[0011] To achieve the above objectives, this utility model provides the following technical solution:

[0012] A confocal multi-well plate for tube forming experiments includes a base plate structure, a receiving hole structure, and a protrusion structure. The receiving hole structure is equidistantly formed on the upper surface of the base plate structure, and the protrusion structure is disposed on the bottom of the inner wall of the receiving hole structure.

[0013] Preferably, the protruding structure is a ring-shaped protruding structure, and the surface of the protruding structure is modified with superhydrophilicity.

[0014] Preferably, the inner wall of the accommodating hole structure is subjected to gradient modification treatment, specifically, the top of the inner wall of the accommodating hole structure is hydrophobically modified, and the lower part of the inner wall of the accommodating hole structure is hydrophilically modified.

[0015] Preferably, the protrusion structure is a micron-sized protrusion structure.

[0016] Preferably, the upper surface of the base plate structure is provided with a crisscrossing groove structure, and the upper edge of the groove structure is provided with a chamfer.

[0017] Compared with the prior art, the beneficial effects of this utility model are:

[0018] 1. The confocal multi-well plate used for tube forming experiments achieves self-leveling of the matrix adhesive through a micron-level annular protrusion array designed at the bottom of the wells combined with superhydrophilic surface modification, so that the standard deviation of the adhesive layer thickness is controlled within ±3μm.

[0019] 2. The confocal multi-well plate used for tube forming experiments adopts a gradient hydrophobic treatment on the sidewalls (hydrophobic at the top / hydrophilic at the bottom) to reduce the liquid level difference to <0.1mm and eliminate halo artifacts in confocal imaging. Attached Figure Description

[0020] Figure 1 This is a schematic diagram of the structure of this utility model;

[0021] Figure 2 This is a cross-sectional view of the structure of this utility model.

[0022] In the diagram: 1. Base plate structure; 2. Receiving hole structure; 3. Protrusion structure; 4. Groove structure. Detailed Implementation

[0023] 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.

[0024] This utility model provides a confocal multi-well plate technical solution for tube forming experiments:

[0025] Example:

[0026] The focusing porous plate mainly includes a base plate structure 1, and a receiving hole structure 2 for holding the substrate is opened at equal intervals on the upper surface of the base plate structure 1. A protruding structure 3 is arranged in an array on the bottom of the inner wall of the receiving hole structure 2.

[0027] Specifically, the protruding structure 3 is a ring-shaped protruding structure, and the surface of the protruding structure 3 is treated with superhydrophilic modification.

[0028] Among them, the inner wall of the accommodating hole structure 2 is treated with gradient modification, specifically, the top of the inner wall of the accommodating hole structure 2 is treated with hydrophobic modification, and the lower part of the inner wall of the accommodating hole structure 2 is treated with hydrophilic modification.

[0029] Specifically, the protrusion structure 3 is a micron-sized protrusion structure.

[0030] The base plate structure 1 has a crisscrossing groove structure 4 on its upper surface, and the upper edge of the groove structure 4 is chamfered. The groove structure 4 is designed to prevent the solution stored inside the porous structure 2 from sloshing and splashing when the entire porous plate is moved, thus effectively preventing cross-contamination.

[0031] In this embodiment, during use, the matrix gel is first filled directly into the cavity of the receiving hole structure 2. After the matrix gel stabilizes, cell suspension is then filled into it. Through the micron-level annular protrusion array designed at the bottom of the hole combined with superhydrophilic surface modification, the matrix gel achieves self-leveling, and the standard deviation of the gel layer thickness is controlled within ±3μm. The sidewall gradient hydrophobic treatment (hydrophobic at the top / hydrophilic at the bottom) reduces the liquid level height difference to <0.1mm, eliminating halo artifacts in confocal imaging.

[0032] Although embodiments of the present invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the present invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A confocal multiwell plate for tube formation assays comprising a base plate structure (1), a well containing structure (2) and a protrusion structure (3), characterized in that: The receiving hole structure (2) is equally spaced on the upper surface of the base plate structure (1), and the protruding structure (3) is set on the bottom of the inner wall of the receiving hole structure (2).

2. The confocal multi-well plate for tube forming experiments according to claim 1, characterized in that: The protruding structure (3) is specifically a ring-shaped protruding structure, and the surface of the protruding structure (3) is modified to be superhydrophilic.

3. A confocal multi-well plate for tube forming experiments according to claim 1, characterized in that: The inner wall of the accommodating hole structure (2) is subjected to gradient modification treatment, specifically, the top of the inner wall of the accommodating hole structure (2) is hydrophobically modified, and the lower part of the inner wall of the accommodating hole structure (2) is hydrophilically modified.

4. A confocal multi-well plate for tube forming experiments according to claim 1, characterized in that: The protrusion structure (3) is specifically a micron-sized protrusion structure.

5. A confocal multi-well plate for tube forming experiments according to claim 1, characterized in that: The upper surface of the base plate structure (1) is provided with a crisscrossing groove structure (4), and the upper edge of the groove structure (4) is provided with a chamfer.

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