Fluorescence-Raman dual-mode nano sensor, preparation method thereof and application of fluorescence-Raman dual-mode nano sensor in mercaptan detection
A nanosensor and dual-mode technology, applied in the field of nanomaterials and life sciences, to achieve high sensitivity, simple and feasible preparation method, and stable properties
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Embodiment 1
[0059] Example 1 Synthesis and Characterization of Gold Nanoflowers (AuNFs)
[0060] 1.25mL 5mM HAuCl 4 The solution was added into 23.75 mL of ultrapure water, and heated at 100° C. for 5 min under vigorous stirring. Quickly add 500 μL of 1% sodium citrate, keep the reaction for 10 min and then cool to room temperature to obtain a gold seed solution. The gold species were characterized by UV-Vis absorption spectrum and TEM electron microscope, the results are as follows figure 1 , where A is the UV-Vis absorption spectrum, and B is the TEM electron microscope.
[0061] Add 500 µL of the gold seed solution to 1 mL of 0.1 M PBS containing 500 µL of 1% PVP. With vigorous stirring, 300 μL of 10 mM HAuCl 4 solution and 300 μL 10mM NH 2 OH-HCl solution was added to the above solution simultaneously. After 2 h, the mixed solution was centrifuged at 5000 rpm for 5 min, and the pellet was resuspended in ultrapure water containing 0.05% Tween 20 to obtain gold nanoflowers (AuNFs)...
Embodiment 2
[0065] Example 2 Functionalization of AuNFs
[0066] 10 μL of 100 μM C1 solution was added to 100 μL of 4nMAuNFs and incubated for 3 h, and then NaCl solution was added dropwise several times to a final concentration of 450 mM. After the mixed solution was incubated overnight, 400 μM mPEG-SH was added and incubated for 30 min. The solution was then centrifuged and the pellet was resuspended in Tris-HCl buffer (50 mM, 100 mM NaCl, pH 7.5) containing 0.01% Tween-20. To make the nanosensors more easily absorbed by the cells, 20 μM TAT was added to the solution and incubated for 30 min. After repeated centrifugation, AuNFs functionalized with C1 (C1-AuNFs) were obtained.
[0067] In order to obtain the maximum loading capacity, the concentrations of the C1 solution were changed to 1 μM, 2 μM, 3 μM, 4 μM, 5 μM, 6 μM and 7 μM to optimize the decrease of the absorbance of the supernatant at 260 nm after centrifugation (ΔA 260 ) demonstrated that the nanoparticles were modified by ...
Embodiment 3
[0072] Embodiment 3 Stability experiment of dual-mode nanosensor:
[0073] (1) Raman spectroscopy characterizes the SERS reproducibility of dual-mode nanosensors:
[0074] Perform Raman spectrum detection on the nanosensor, and collect Raman spectra of no less than 20 points to evaluate its signal reproducibility. The result is as Figure 6 shown.
[0075] (2) UV-Vis spectroscopy to characterize the salt stability of the dual-mode nanosensor:
[0076] The prepared dual-mode nanosensor was centrifuged and resuspended in PBS buffer containing different NaCl concentrations (100mM, 300mM, 500mM, 700mM), vortexed and mixed, and after standing for 10min, the double-mode sensor was detected by ultraviolet-visible absorption spectrometer. Modal nanosensor spectral changes. The result is as Figure 7 -A shown.
[0077] (3) Fluorescent experiments to evaluate the ribozyme stability of dual-mode nanosensors:
[0078] There are a variety of enzymes in the cell, which can cut nuclei...
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