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.
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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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