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A micro-nano composite structure surface matching the size of circulating tumor cells, its preparation method and its application

A micro-nano composite structure, tumor cell technology, applied in micro-structure technology, micro-structure devices, manufacturing micro-structure devices and other directions, can solve the problems of limiting the capture efficiency of the substrate, neglecting the size of target cells, complex preparation, etc. Simple preparation and high capture efficiency

Active Publication Date: 2020-07-17
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method also has the disadvantages of high cost and complicated preparation, and in the process of designing the micro / nanostructured substrate, the size of the target cells is often ignored, so that the cells cannot fully contact the micro / nanostructured substrate, which limits the size of the substrate. capture efficiency

Method used

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  • A micro-nano composite structure surface matching the size of circulating tumor cells, its preparation method and its application
  • A micro-nano composite structure surface matching the size of circulating tumor cells, its preparation method and its application
  • A micro-nano composite structure surface matching the size of circulating tumor cells, its preparation method and its application

Examples

Experimental program
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Effect test

Embodiment 1

[0034] In this example, MCF-7 cells and Hela cells are used as cells to be captured and reference cells (MCF-7 cells have the membrane antigen EpCAM on the surface, but Hela cells do not). Since the sizes of the two are mainly distributed in 10-20 μm, so, In this embodiment, a circular hole array is selected as the mask plate. The array is arranged in a square, with a period of 20 μm and a pore size of 10 μm, to prepare a columnar array arranged in a square, and capture the above two types of cells on the surface of the micro-nano composite structure. The invented capture system is further elaborated and verified. The specific steps for efficiently capturing circulating tumor cells on the surface of the micro-nano composite structure are as follows:

[0035] (1) Construction of micro-nano composite structure surface

[0036] Sonicate silicon wafers with acetone, chloroform, ethanol, and water at a power of 70W for 5 minutes in sequence. After blowing dry with nitrogen, spin t...

Embodiment 2

[0049] The micro-nano composite structure capturing MCF-7 cells described in Example 1 was soaked in a phosphate buffer solution containing 2.5% glutaraldehyde at 4° C. overnight to fix the captured cells. Subsequently, the cells were dehydrated with a series of ethanol aqueous solutions with different volume fractions (0, 30%, 50%, 70%, 90%, 100%) with a certain gradient, and dried at room temperature. The 10nm gold film was sputtered, and the fixed MCF-7 cells were characterized by a Hitachi (SU8020 FE-SEM) scanning electron microscope, and the results were as follows Figure 4shown. The cell body is trapped between the four pillars and interacts with the micron-scale array, and a large number of pseudopodia protrude from the position where the cell contacts each pillar, indicating that the micro-nanostructure on the cell surface can interact with the micro-nanostructure on the pillar surface . Therefore, the micro-nano composite structure matching the size of circulating ...

Embodiment 3

[0051] The micro-nano composite structure matching the size of circulating tumor cells described in Example 1 was named PDMS-1. And using the same method as described in Example 1, replace the photolithography template, and obtain two kinds of and cycle with a period of 10 μm, a column width of 5 μm, a column height of 10 μm and a period of 60 μm, a column width of 30 μm, and a column height of 10 μm. The tumor cell size-mismatched structures were named PDMS-2 and PDMS-3, respectively. Use the same method in Example 1 to carry out metal-catalyzed etching on blank silicon wafers to obtain silicon nanowire arrays, ultrasonically pulverize silicon nanowires, and pour PDMS prepolymers, solidify, cool and peel off, and obtain rough PDMS surface structures, named as RPDMS. Scanning electron microscope (Hitachi, SU8020FE-SEM) photos of the four PDMS structures are shown in Figure 5 shown. Figure 5 (a)-(d) correspond to the structures of RPDMS, PDMS-1, PDMS-2, and PDMS-3, respect...

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Abstract

The invention discloses a micro-nano composite structure surface matched with circulating tumor cell size, a preparation method and application thereof, and belongs to the technical field of the biologics. The preparation method comprises the following steps: preparing a micro-nano composite structure array matched with a to-be-detected tumor cell size on the surface of the substrate, and then fixing specific antibody on the tumor cell surface on the surface of the structure area, utilizing the structure array to synergistically effect with the specific antibody to realize the efficient and specific capturing on the circulating tumor cell. The theory of designing the micron / nanometer structure is designed according to the target cell size can be further used for detecting other cells, thereby providing a new path for the preparation of the biological detection substrate. The substrate prepared through the invention can efficiently and specifically capture the circulating tumor cell, and has the features of being simple in preparation and low in cost; the captured cell has good survival efficiency, and a new path is provided for the preparation of the biological detection substrate.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a surface of a micro-nano composite structure matching the size of circulating tumor cells, a preparation method and its application in efficiently capturing circulating tumor cells. Background technique [0002] Malignant tumor is one of the three major diseases that cause human death, and more than 90% of the deaths are caused by metastatic tumors. Tumor cells or cell clusters break away from the original tumor location, enter the blood or lymph fluid, and then spread to other organs, providing the possibility for tumor metastasis. This makes it particularly important to detect these tumor cells that are free in blood or lymph. In 1869, Professor Ashworth (Aust.Med.J., 1869,14,146-147) discovered circulating tumor cells (Circulating Tumor Cells) in the peripheral blood of cancer patients for the first time through a thorough comparison with different types of tumor cel...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B81C1/00B82Y40/00G01N33/574
CPCB81C1/00373B81C1/00404B82Y40/00G01N33/57484
Inventor 吕男王中舜徐大任金悦霍博王亚磊付学奇
Owner JILIN UNIV
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