Microarray chip without solid wall based on LSPR (Localized Surface Plasmon Resonance) and application thereof

A technology of microarray chips and substrates, which is applied in the measurement of scattering properties, etc., can solve the problems of low-cost detection and expensive detection instruments, etc., and achieve the effects of low cost, wide application prospect, and easy operation.

Active Publication Date: 2013-10-02
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the method based on single particle detection often relies on the dark field scattering spectrum microscope that can measure

Method used

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  • Microarray chip without solid wall based on LSPR (Localized Surface Plasmon Resonance) and application thereof
  • Microarray chip without solid wall based on LSPR (Localized Surface Plasmon Resonance) and application thereof
  • Microarray chip without solid wall based on LSPR (Localized Surface Plasmon Resonance) and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1. Preparation of LSPR-based microarray chip base without solid walls

[0049] 1. Preparation of the mold (i.e. array mold) used to prepare the hydrophilic-hydrophobic array

[0050] The array mold is a flat plate with round through holes; the round through holes are arranged in an array of 8×3 (that is, 8 rows in the horizontal direction and 3 rows in the vertical direction). , and the distance between the centers of each circle is 9mm. The plate material is polydimethylsiloxane (PDMS).

[0051] The specific production process is as follows:

[0052] 1) First, the mold pool used to form the array mold is obtained by laser cutting method, as follows:

[0053] Use AutoCAD software to draw, draw a rectangle with a width of 100mm and a length of 130mm, which is recorded as pattern 1; then draw a rectangle with a width of 100mm and a length of 130mm, and draw a rectangle of 80mm×110mm (width×length) in this rectangle, record for pattern 2. According to the patt...

Embodiment 2

[0071] Embodiment 2, preparation of LSPR array chip without solid wall with affinity molecules

[0072] 1. Preparation of LSPR chip without solid wall for detection of adenosine triphosphate (ATP)

[0073] A non-solid wall LSPR chip with high sensitivity and specific detection of adenosine triphosphate (ATP) was obtained by competitive replacement method. The specific preparation method is as follows:

[0074] A) The specific nucleic acid aptamer 5'-SH-(CH 2 ) 6 -TTCTCCCCCAGGTAA-3' (its nucleotide sequence is shown in Sequence Listing Sequence 1) and oligoethylene glycol (11-mercaptoundecyl) tri(ethylene glycol) are mixed in PBS buffer (pH7. In 4), the concentrations reached 1 μM and 5 μM respectively to obtain aptamer / oligoethylene glycol mixed solution. Take 10 μL of the aptamer / oligoethylene glycol solution and drop it on the hydrophilic area of ​​the LSPR glass slide without solid wall, let it stand at room temperature for more than 3 hours in a humid chamber, wash it w...

Embodiment 3

[0082] Embodiment 3, the preparation of detection adapter

[0083] According to the preparation method of the mold pool of the array mold described in step 1 of embodiment 1, make a detection adapter that matches the detection equipment and can fix the chip described in embodiment 2, and its size can be determined according to the size of the position where the sample is placed by the detection equipment, The following is just an example of a detection adapter suitable for the microplate detection system SpectraMax M5 / M5e:

[0084] Use AutoCAD software to draw, draw a rectangle of 86mm×128mm (width×length), and draw four rectangles of 71.2mm×23mm in this rectangle, and the distance between them is 4mm, which is recorded as pattern 1; then draw out Draw a rectangle of 86mm×128mm (width×length), and draw four rectangles of 26mm×76mm in this rectangle. The distance between them is 1.55mm, which is marked as pattern 2. The size of the PDMS array template is an array template with...

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Abstract

The invention discloses a microarray chip without solid walls based on LSPR (Localized Surface Plasmon Resonance) and application thereof. The substrate of the microarray chip is in a hydrophilic and hydrophobic mode and comprises a base, a plurality of hydrophilic regions and hydrophobic regions, wherein the hydrophilic regions are arranged on the base; the hydrophobic regions separate the hydrophilic regions; the surfaces of the hydrophilic regions are provided with metal nanometer material layers with local area surface plasma resonance attributes; the outer regions of the hydrophilic regions are not provided with the solid walls; the surfaces of the hydrophobic regions are provided with hydrophobic material layers. The microarray chip disclosed by the invention can be obtained by combining a peculiar affinitive molecule for detecting a substance to be detected on the substrate provided by the invention. The microarray chip disclosed by the invention has the advantages of easiness and convenience for use, low detection cost, accurate result, reusability, and the like, can be used for detecting multiple substances, such as micromolecules, heavy metal ions, proteins, bacteria, viruses, and the like, and has wide application prospect in the fields of medical health, environmental monitoring, scientific experiments, and the like.

Description

technical field [0001] The invention relates to an LSPR-based microarray chip without solid walls and its application. Background technique [0002] Due to their unique optical properties, noble metal nanomaterials are widely used in sensing, thermotherapy, imaging and other fields. Localized surface plasmon resonance (LSPR) is an important optical property of noble metal nanostructures. When light of a certain frequency is incident on the surface of nanoparticles, the light couples with the free electrons on the surface of nanoparticles to generate resonance. This resonance is macroscopically manifested as spectral selectivity. Absorption, this resonant mode is very sensitive to changes in the refractive index of the medium in the region surrounding the metal nanostructure. LSPR-based nanosensors have proven to be an effective platform for the quantitative detection of biomarkers or chemicals. With the help of the new characteristics of nanomaterials, LSPR not only has th...

Claims

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Application Information

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IPC IPC(8): G01N21/55
Inventor 杜亚楠谢丽萍鄢晓君
Owner TSINGHUA UNIV
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