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Optical-interference-free Raman labeling probe and preparation method and application thereof

A technology for labeling probes and Raman, which is used in Raman scattering, material analysis by optical means, scientific instruments, etc. It can solve the problems of weak signal intensity of spontaneous Raman scattering, inability to become a general-purpose instrument for biological imaging, and sample damage. , to achieve the effect of shortening biological imaging time, avoiding overlapping, and eliminating interference peaks

Active Publication Date: 2016-08-03
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the wide application of alkyne signal molecules is still severely limited by the weak intensity of spontaneous Raman scattering signals. In order to detect extremely weak alkyne-based Raman scattering signals, people have to increase the exposure time or use high-intensity lasers. , which may cause damage to the sample; in recent years, some people have tried to use Stimulated Raman scattering (Stimulated Ramanscattering, SRS) technology for related research, but the expensive instrument price makes people daunting. The limited availability of the instrument also prevents it from becoming a general-purpose instrument in the field of bioimaging

Method used

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  • Optical-interference-free Raman labeling probe and preparation method and application thereof
  • Optical-interference-free Raman labeling probe and preparation method and application thereof
  • Optical-interference-free Raman labeling probe and preparation method and application thereof

Examples

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Effect test

Embodiment 1

[0044] Example 1: Preparation of narrow-band single-peak Raman signal molecule

[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, t...

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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Abstract

The invention designs a series of signal molecules with Raman scattering peaks in the wave number area of 1800-2400 cm<-1> by using a sulfydryl phenylacetylene as a body structure, selects gold and silver nano-particles as reinforcing substrates of Raman signals, enables alkyne signal molecules with sulfydryls to be self-assembled on the surfaces of the reinforcing substrates, uses mercaptoacetic acid to optimize SERS signals, then selects a light-transmitting and hydrophilic package material rich in active reaction groups for probe package and finally grafts specific biological targeting functional molecules to an outer layer of the package material .A probe has strong optical response to Raman scattering, adopts narrow-band unimodal transmission, is free of optical background interferences and has the great significance in the biological imaging field of simultaneous marking of multiple components .The technology can extend to design and development of a no-raster ultra-fast Raman imager using a bandpass filter as a beam splitting device and using a photomultiplier tube (PMT) or an avalanche photodiode (APD) as a detecting device, overcome the technical bottlenecks of Raman imaging point-by-point scanning and slow imaging and fills the blank in the market of optical imaging instruments .

Description

technical field [0001] The invention belongs to the technical field of biological imaging, and relates to a preparation method of a novel alkyne-encoded and biologically targeted surface-enhanced Raman scattering (SERS) active nano-labeled probe. More importantly, it involves the design and development of a grating-free ultrafast Raman imager with a bandpass filter as the spectroscopic device and a photomultiplier tube (PMT) or avalanche photodiode (APD) as the detection device. Background technique [0002] Biological imaging (such as live cell imaging) technology allows scientists to observe the internal structure and physiological processes of living organisms in real time and dynamically, revolutionizing the way biologists study the mechanism of biological activity. Among the many biological imaging techniques, fluorescent labeling imaging is the most widely used and mature. It uses signal probes that can absorb light of specific wavelengths and emit light of longer wav...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/65
CPCG01N21/658
Inventor 沈爱国陈勇任家强胡继明
Owner WUHAN UNIV
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