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Addressable sample plate and application thereof in single cell microscopic imaging

A sample plate, single-cell technology, applied in measurement devices, material analysis by optical means, instruments, etc., can solve problems such as difficulty in positioning single cells, and achieve the effect of reducing difficulty

Inactive Publication Date: 2016-11-23
INST OF CHEM CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to overcome the technical problem of single cell location difficulty existing in the combined analysis of topography imaging and component imaging, and to provide an addressable sample plate that can easily and quickly find designated cells in a large area and the Application of Sample Plates in Single-Cell Microscopic Imaging

Method used

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  • Addressable sample plate and application thereof in single cell microscopic imaging
  • Addressable sample plate and application thereof in single cell microscopic imaging
  • Addressable sample plate and application thereof in single cell microscopic imaging

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] This example is used to illustrate the preparation and characterization of the addressable sample plate of the present invention.

[0044] (1) Preparation of chromium template

[0045] Using EBL electron beam exposure technology to write as Figure 7 On-board graphics shown (A, B, C, D, 1, 2, 3, 4, 4 × 4 arrays, each cell size is 50 × 50 μm 2 ), and corrode the chromium film to make a chromium template.

[0046] (2) Preparation of sample plate

[0047] The template obtained in the above step (1) is used for ultraviolet exposure to make a patterned photoresist mask. 1×1cm under the protection of the mask 2 ICP silicon etching is performed on the double-sided polished silicon wafer, and the width and depth required for etching are both 3 μm. Wash three times ultrasonically with 5 mL of acetone, and blow dry with nitrogen to obtain a sample plate.

[0048] (3) Characterization of the sample plate

[0049] Observation of the obtained sample plate under a microscope (...

Embodiment 2

[0051] This embodiment is used to illustrate that the sample plate prepared in Example 1 is used for SIMS-Confocal combined imaging research on ruthenium compound 1 (structural formula such as figure 2 A) Method of spatial distribution in MCF-7 cells.

[0052] (1) Culture and lyophilization of MCF-7 cells

[0053] Place the sample plate in the center of the bottom of the cell culture dish for laser confocal, digested MCF-7 cells in 10 4 cells / cm 2 The concentration was cultured in a petri dish for 24 hours, and the medium containing 100 μM ruthenium compound 1 was added to incubate for 24 hours, the medium was removed, and the culture medium containing 1 μg / mL Hoechst 33342 was incubated in a 37°C incubator for 10 minutes. Remove the medium, and then incubate for 20 min in a 37°C incubator with a culture medium containing 5 μM cell membrane red fluorescent dye. The medium was removed, and the cells were treated with NH at a concentration of 150 mM 4 COOCH 3 (pH 7.4) buff...

Embodiment 3

[0064] This embodiment is used to illustrate that the sample plate prepared in Example 1 is used for SIMS-Confocal combined imaging research on ruthenium compound 2 (structural formula such as figure 2 B) Means of spatial distribution in MCF-7 cells.

[0065] (1) Culture and lyophilization of MCF-7 cells

[0066]Place the sample plate in the center of the bottom of the cell culture dish for laser confocal, digested MCF-7 cells in 10 4 cells / cm 2 The concentration was cultured in a petri dish for 24 hours, and the medium containing 100 μM ruthenium compound 2 was added to incubate for 24 hours, the medium was removed, and the culture medium containing 1 μg / mL Hoechst 33342 was incubated in a 37°C incubator for 10 minutes. Remove the medium, and then incubate for 20 min in a 37°C incubator with a culture medium containing 5 μM / L cell membrane red fluorescent dye. The medium was removed, and the cells were treated with NH at a concentration of 150 mM 4 COOCH 3 (pH 7.4) buff...

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Abstract

The invention belongs to the field of microscopic imaging, and discloses an addressable sample plate and application thereof in single cell microscopic imaging. N n * m arrays are carved on the addressable sample plate, and the cell size of the array is 100 [mu]m<2>-4 * 10<4> [mu]m<2>. The invention also discloses the application of the sample plate in single cell microscopic imaging and a single cell microscopic imaging method. The single cell microscopic imaging method comprises fixing of cells on the sample plate, labeling with a fluorescent dye, and respective performing of fluorescence imaging analysis and mass spectrum imaging analysis of the cells in the same cell. When the addressable sample plate is used for single cell microscopic imaging analysis, the addressable sample plate is advantageous in clear and accurate positioning of the same cell in a large area range, and the difficulty of positioning of the same cell when a variety of analytical instruments are combined in use can be reduced. In addition, the sample plate does not impede cell attachment and growth, so that the sample plate has no adverse effect on analytical results.

Description

technical field [0001] The invention belongs to the field of microscopic imaging, and in particular relates to an addressable sample plate and the application of the sample plate in single-cell microscopic imaging. Background technique [0002] So far, the research on the mechanism of metal antitumor drugs inhibiting tumor cells is not deep enough. Moreover, the interaction mechanism between drugs and biological target molecules studied at the molecular level cannot reflect the real situation in cells. Therefore, the study of the spatial distribution of drugs in cells is of great significance for in-depth understanding of the mechanism of action of anti-tumor drugs and further optimization of their molecular design, which is also one of the most challenging topics in the field of drug development. [0003] At present, the main methods for studying the distribution of drugs in cells are fluorescent labeling imaging and mass spectrometry imaging. The fluorescent labeling met...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/01G01N27/64
Inventor 汪福意刘素彦吴魁罗群赵耀
Owner INST OF CHEM CHINESE ACAD OF SCI
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