A Raman-labeled probe without optical interference and its preparation method and application
A labeling probe and Raman technology, applied in Raman scattering, material analysis through optical means, scientific instruments, etc., can solve the problem of weak signal intensity of spontaneous Raman scattering, sample damage, failure to become a general-purpose instrument in the field of biological imaging, etc. problems, to achieve the effects of shortening biological imaging time, eliminating interference peaks, and avoiding overlapping
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Embodiment 1
[0044] Example 1: Preparation of narrow-band single-peak Raman signal molecules
[0045] Alkyne narrow-band single-peak Raman signal molecular design: the present invention uses p-mercaptophenylacetylene as the main structure, and changes the displacement of the alkynyl Raman scattering signal by replacing the alkynyl terminal substituent group, and uses density functional theory to calculate the A series of predictions and theoretical studies have been carried out on the relationship between the structure of the signal molecule and the Raman shift, and the chemical structure and its Raman shift in the table below have been obtained, which proves that when the group is changed at the end of the alkyne group of p-mercaptophenylacetylene, it can effectively Modulation of Raman Signal Shifts of Narrowband Singlet Signaling Molecules.
[0046]
[0047] Spectrum optimization of signal molecules: Using silver-coated gold nanoparticles of a certain size as the enhanced substrate, ...
Embodiment 2
[0050] (1) AuNPs-labeled molecules: Take three 10 mL silver-coated gold nanosols, and add 10 μL of 1 mM narrow-band unimodal signal molecules B1, B2, and B3 in tetrahydrofuran respectively.
[0051] (2) Optimization of thioglycolic acid: After the above solution was allowed to stand for 3 hours, 100 μL of an aqueous solution of thioglycolic acid with a concentration of 10 mmol / L was added. (This step is only required when the alkynyl terminal of the signal molecule is not substituted, for example, B1 is used as the signal molecule).
[0052] (3) Protective shell layer: After standing still for 3 hours, continue to add 1 mL of polycyclic aromatic hydrocarbon (PAH) aqueous solution with a concentration of 0.1 wt‰.
[0053] (4) Biological functional molecule connection: 10 μL of 10 mM 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC·HCl) aqueous solution and 20 μL of 1mM N-hydroxysuccinimide (NHS) aqueous solution, let stand to activate the carboxyl group, add 20...
Embodiment 3
[0057] Take 10mL particle concentration is about 10 12 silver-coated gold nano-sol per mL, add 20 μL of B3 signal molecules with a concentration of 1mM to it, and then continue to add 1mL of polycyclic aromatic hydrocarbon (PAH) aqueous solution with a concentration of 0.1wt‰ after standing for 12 hours. SERS probe.
[0058] Take 10 mL of an aqueous solution in which lily pollen is evenly dispersed, and add 1 mL of SERS probe to it, mix well on a shaker, and freeze-dry the pollen solution to remove water.
[0059] Prepare a high-concentration sucrose solution and drop a small amount onto the glass slide. After drying, a uniform and transparent sucrose layer is formed on the surface. Take a small amount of pollen grains and sprinkle them on the sucrose layer and gently blow air to apply pressure, so that the pollen can be firmly adhered to the surface of the sucrose, and the Raman imaging experiment can be performed.
[0060] Based on the flavonoids in pollen at 1580cm -1 Th...
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