Preparation method of function surface strength laman scattering probe

A surface-enhanced Raman, functional technology, used in Raman scattering, biochemical equipment and methods, and microbial determination/inspection. It can solve problems such as narrow excitation spectrum, low fluorescence intensity, and background fluorescence flooding, avoiding the Measurement error, the effect of improving reliability

Inactive Publication Date: 2007-12-19
SOUTHEAST UNIV
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Problems solved by technology

Fluorescent probes have been widely used due to their advantages of fast detection speed and simplicity, but they have disadvantages such as low fluorescence intensity, narrow excitation spectrum, and easy to be photobleached.
Raman probes are detected by Raman spectroscopy, which has clear spectral features and can provide rich information on molecular structures, but the intensity of ordinary Raman scattering signals is very low and is easily overwhelmed by background fluorescence, so detection is difficult

Method used

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  • Preparation method of function surface strength laman scattering probe
  • Preparation method of function surface strength laman scattering probe
  • Preparation method of function surface strength laman scattering probe

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1, preparation of functional SERS probe

[0044] In the first step, first prepare silver colloid solution according to the method described by Lee and Meisel: under ice bath condition, make 50mL concentration 5×10 -3 Add 150mL of silver nitrate solution of M to a concentration of 2×10 -3 M in sodium borate solution and mix well, and stir at room temperature for 1 hour. Obtain a yellow-green transparent solution, that is, a silver colloid solution, for use.

[0045] In the second step, the concentration is 10 -5 The methanol solution of 2-aminophenol sulfur of M and the silver colloid solution prepared in the first step were uniformly mixed in a volume ratio of 3:1000, and reacted for 20 minutes. 2-Aminophenol-sulfur molecules are easily intercalated on the surface of silver nanoparticles in silver colloid through -HS bonds. Silver nanoparticles labeled with 2-aminophenol sulfur molecules were separated from solution by centrifugation. The rotational spee...

Embodiment 2

[0046] Example 2, Functional SERS Probe Used for pH Sensing in Pancreatic Cancer Cells

[0047] In the first step, the response curve of the SERS probe to pH is measured. The functional SERS probe solution was added to the buffer solution of pH3.0~pH8.0, the pH value interval was 1.0. After 10 minutes, the mixed solution was dropped onto the silicon chip and fixed on the carrier of the Raman confocal microscope. on the object stage to measure its SERS spectrum. Repeat the measurement 10 times corresponding to each pH value, and calculate the 1505cm in each SERS spectrum -1 at 1390cm -1 The ratio of signal strength at the place, that is, I 1505 / I 1390 , taking the average of 10 experimental results. Taking pH as the abscissa, I 1505 / I 1390 Draw the response curve of the SERS probe to pH as the ordinate, as shown in Figure 1-3. The probe has a high acid detection limit, and the linear response range is 3.0-6.0.

[0048] In the second step, pancreatic cancer cells were...

Embodiment 3

[0050] Example 3, Functional SERS Probe Used for Multicomponent Detection in Pancreatic Cancer Cells

[0051] In the first step, pancreatic cancer cells were cultured in culture medium (37°C, 5% CO 2 ). After 24 hours, the SERS probe solution was added to the culture medium in a volume ratio (3:1), shaken gently, and placed in the incubator again. At this time, the SERS probe enters the interior of the cell by being phagocytized by the cell. One hour later, the medium was discarded, and the cells were gently washed 3 times with phosphate buffered saline (PBS, 7.4) to remove the SERS probes remaining in the medium that were not phagocytized by the cells. stand-by.

[0052] In the second step, the cells treated in the first step were placed in phosphate buffered saline (PBS, 7.4), and fixed on the stage of a confocal Raman microscope. Select a single-cell region to detect the SERS spectrum in the cell, as shown in Figure 3. In addition to the original signal of the probe, a...

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Abstract

A method for preparing functional Roman scattering probe with intensified surface includes mixing 10-5 M of sensitive material using (-NH2) as PH sensitive group with silver glue solution in volume ratio of 3:1000-300:1000 uniformly then adding o.1M of sodium chloride solution in as per volume ratio of 3:1-10:1 for obtaining functional Roman scattering probe in size of 20-50nm with intensified surface.

Description

technical field [0001] The invention relates to the field of laser Raman spectrum detection, in particular to a preparation method of a functional surface-enhanced Raman scattering probe and its application to intracellular pH sensing and intracellular multi-component detection. Background technique [0002] Due to the important role of optical probes in cell imaging and real-time acquisition of intracellular information, people have paid more attention to the preparation of optical probes and their application at the single-cell level in recent years, and developed various types and structures of optical probes. Probes are used to study cell morphology or detect intracellular components, etc. There are two types of optical probes commonly used: fluorescent probes and Raman probes. Fluorescent probes have been widely used due to their advantages of fast detection and simplicity, but they have disadvantages such as low fluorescence intensity, narrow excitation spectrum, and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/65C12Q1/04
Inventor 王著元崔一平恽斌峰胡国华
Owner SOUTHEAST UNIV
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