High-flux ultra-sensitive detection dot array enhancement chip
An array chip, high-throughput technology, applied in measuring devices, nanotechnology for sensing, material analysis through optical means, etc., can solve problems such as high detection limit, single detection target, and low detection sensitivity
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Embodiment 1
[0034] The preparation of embodiment 1 chip
[0035] Such as figure 1 with figure 2 As shown, the chip preparation process of the present invention is as follows:
[0036] (1) Cleaning of glass slides and microspheres: ultrasonically clean the slides with absolute ethanol and ultrapure water respectively for 30 minutes; dry the cleaned slides and clean them in a plasma cleaner for 30 minutes. for 15 minutes; store the cleaned slides in ultrapure water.
[0037] (2) The microspheres are arranged at fixed points on the glass interface, that is, use a pipette gun to draw 50 microliters of the microsphere solution with a concentration of 1mol / L and drop it on the slide. When the microsphere solution diffuses to the bottom of the slide, the bottom of the slide Rotate 180° to the top to reflux part of the microsphere solution to form a single-layer microsphere lattice with a larger area;
[0038] (3) Coating the glass slide with aluminum in a magnetron sputtering apparatus to f...
Embodiment 2
[0040] Example 2 Raman signal detection of gold rods (AuNR) and gold balls (AuNP) on the chip
[0041] The synthesized gold rods (AuNR) and gold balls (AuNP) were deposited into the nanoholes, and the gold rods self-assembled in the holes to form a signal-enhancing "hot spot" lattice array, and then 200 microliters of Raman probe molecule hemoglobin was added and shaken by ultrasound And detect the Raman signal, and calculate the lattice enhancement effect. Test results such as Figure 5 As shown, the results show that, compared with pure hemoglobin molecules, the AuNR and AuNP lattices at wavenumber 600cm -1 and 1121cm -1 All have enhanced signals, and the AuNP has a wavenumber of 1600cm -1 There is also an enhanced signal. After calculation, the enhanced effect of this chip can reach 10 2 times. It can be seen that the chip has good signal enhancement performance.
Embodiment 3
[0042] Example 3 Detection of pathogenic microorganism microRNA
[0043] Gold rod or golden cross coupling DNA steps:
[0044] 1. Add equal volumes of 100uM ssDNA and 1mM TCEP to the PCR tube, reaction conditions: keep it in the dark for 3 hours
[0045] 2. Absorb 0.1M gold nanorods or gold cross 1mL;
[0046] 3. Add the volume of SDS-HCL (pH=3) equal to that of step 1 to step 2;
[0047] 4. Shake at room temperature for 12 hours at a speed of 1000rpm;
[0048]5. Centrifuge at 1000rpm for 10min, absorb the supernatant, and add 0.005M CTAB to the precipitate for later use.
[0049] Gold nanocrosses linked to DNA2, whose ends are modified with a cy5 fluorescent dye, were deposited into the holes. In addition, DNA1 is connected to the gold rod, and this DNA strand is complementary to part of DNA2 base pairing. At this time, the cy5 fluorescent dye does not emit light and is in a quenched state; when pathogenic microorganism miRNA-21 is added, due to the complementary base pai...
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Abstract
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