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Chip for capturing, counting and lossless release of circulating tumor cells and application of chip

A tumor cell and chip technology, applied in the field of functional nanomaterials and biological detection, can solve the problems of insufficient strategies for sensing, detecting and counting circulating tumor cells, difficulty in non-destructive release of circulating tumor cells, and low abundance of circulating tumor cells, achieving excellent space Positioning ability, obvious affinity performance, effect of improving capture ability and effect

Active Publication Date: 2022-07-12
NANJING UNIV OF POSTS & TELECOMM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Purpose of the invention: In view of the extremely low abundance and heterogeneity of circulating tumor cells, the difficulty of releasing circulating tumor cells without damage, and the lack of current strategies for detecting and counting circulating tumor cells, the present invention proposes a method for capturing circulating tumor cells , Counting, Losslessly Released Chips

Method used

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  • Chip for capturing, counting and lossless release of circulating tumor cells and application of chip
  • Chip for capturing, counting and lossless release of circulating tumor cells and application of chip
  • Chip for capturing, counting and lossless release of circulating tumor cells and application of chip

Examples

Experimental program
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Effect test

Embodiment 1

[0074] Example 1 Preparation of chips for capturing, counting and non-destructive release of circulating tumor cells

[0075] The first step is to use the oblique angle deposition method to prepare the evaporation coating technology ( [3] Liu Y J, Chu H Y, Zhao YP. Silver Nanorod Array Substrates Fabricated by Oblique Angle Deposition: Morphological, Optical, and SERS Characterizations [J]. The Journal ofPhysical Chemistry C, 2010, 114(18): 8176-8183). A 1 cm × 1 cm glass slide was placed in a vacuum deposition chamber, and 20 nm Ti, 200 nm Ag film, and 2000 nm film were sequentially deposited at the rates of 0.2 nm / s, 0.3 nm / s, and 0.3 nm / s, respectively. Ag film to form silver nanorods with morphologies such as figure 1 shown. The whole evaporation process is under pressure -6 Torr under high vacuum conditions.

[0076] In the second step, PDMS small holes with a diameter of 4 mm and a height of 1 mm were prepared and pasted on the silver nanorod array substrate to obtai...

Embodiment 2

[0083] Example 2 Characterization of the working mechanism of the chip for capturing, counting and non-destructive release of circulating tumor cells

[0084] 1. To characterize the working mechanism of the chip prepared in Example 1 for capturing, counting, and nondestructively releasing circulating tumor cells. like figure 2 As shown in (b), the constructed double-tetrahedral DNA probe is covalently bound to the silver nanorod through three thiol groups on the bottom surface of the base tetrahedral DNA to form a biorecognition interface with nano-rough surface features. .

[0085] When capturing and releasing circulating tumor cells, first, a Zn ion-responsive DNAzyme-aptamer chain (which can specifically target and bind to cell surface membrane proteins) was co-coated with circulating tumor cells (human gastric cancer cell SGC-7901 as an analytical model). After incubation, the aptamer is modified to the surface of circulating tumor cells by specific binding of the aptam...

Embodiment 3

[0093] Example 3 Characterization of cell-on-chip capture performance for the capture, counting, and non-destructive release of circulating tumor cells

[0094] 1. In the case of ensuring that the Zn ion-responsive DNAzyme-aptamer chain is in excess relative to the number of cells, the Zn ion-responsive DNAzyme-aptamer chain was co-incubated with the target cell SGC-7901 to obtain Zn ion-responsive DNAzyme- Aptamer chain-labeled SGC-7901 cells.

[0095]2. Co-incubate the chip with SGC-7901 cells bound with Zn ion-responsive DNAzyme-aptamer chains to capture and separate cells. In order to determine the optimal cell capture time, the cell capture performance of the nanobiointerface with different incubation times from 0 to 50 min was experimentally investigated. Figure 4 The middle (a) is the fluorescence imaging characterization of the captured cells for 0~50 min, Figure 4 The middle (b) is the corresponding capture efficiency statistics. With the increase of incubation ti...

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Abstract

The invention discloses a chip for capturing, counting and lossless release of circulating tumor cells and application of the chip. Ditetrahedron DNA probes of the chip are assembled to a silver nanorod substrate to form a biological recognition interface with nano rough surface characteristics; the Zn ion response type DNAzyme-aptamer chain is mixed with a to-be-detected sample, and is bound to the surface of a cell through specific binding of an aptamer and membrane protein of a target cell in the to-be-detected sample. A sample to be detected and the chip are co-cultured, target cells are captured by the chip through hybridization of a Zn ion response type DNAzyme-aptamer chain and a Zn ion response type DNAzyme substrate-connecting chain, and separation from a complex sample is realized; the cells captured by the chip are counted through fluorescent molecules modified on a fluorescence imaging Zn enzyme-aptamer chain; by adding Zn ions to the chip, DNAzyme nucleic acid cleavage is implemented, and lossless release of cells from the chip is realized.

Description

technical field [0001] The invention belongs to the fields of functional nanomaterials and biological detection, and in particular relates to a chip for capturing, counting and nondestructively releasing circulating tumor cells and its application. Background technique [0002] Among cancer deaths, more than 90% of cancer patients die of tumor metastasis and recurrence. Due to the invasive and metastatic properties of tumors, tumor cells are able to slough off the tumor in situ or its metastases into the blood circulation to form circulating tumor cells. In recent years, circulating tumor cells have been recognized as important tumor markers, which are closely related to the mechanism of tumor metastasis. Detecting and counting circulating tumor cells in peripheral blood has important guiding significance for tumor occurrence, recurrence and prognosis evaluation. However, the content of circulating tumor cells in peripheral blood is extremely low, usually only a few or doze...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/569G01N33/543G01N33/58C12Q1/6816
CPCG01N33/574G01N33/56966G01N33/54346G01N33/582C12Q1/6816C12Q2563/107C12Q2521/345C12Q2563/157Y02A50/30
Inventor 宋春元汪联辉李进翔
Owner NANJING UNIV OF POSTS & TELECOMM
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