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Method for detecting animal active unicellular sample by surface reinforced Raman spectrum

A surface-enhanced Raman and spectroscopic detection technology, applied in the field of biomedicine, can solve the problems of micropores in the cell membrane, decrease, and the cell membrane hinders the permeability of particles, and achieves the effect of high transfer efficiency and a simple method.

Inactive Publication Date: 2009-07-15
FUJIAN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when the cell membrane is stimulated by an electric pulse with an intensity of KV / cm and a duration of μs to ms, micropores will appear in the cell membrane, resulting in a decrease in the ability of the cell membrane to hinder particle penetration.

Method used

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  • Method for detecting animal active unicellular sample by surface reinforced Raman spectrum
  • Method for detecting animal active unicellular sample by surface reinforced Raman spectrum
  • Method for detecting animal active unicellular sample by surface reinforced Raman spectrum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Dissolve 9mg of silver nitrate in 50ml of deionized water, heat to 100°C to make it boil, then add 1ml of 3% trisodium citrate dropwise, stir rapidly, and keep boiling for 2 hours after the dropwise addition. After the silver sol was naturally cooled, the layers were centrifuged in a centrifuge, the supernatant was discarded, and the concentrated silver sol in the lower layer was sealed and stored at room temperature away from light for later use.

[0022] The test sample epithelial tumor cells (A431) to be tested are first prepared with commercially available RPMI 1640 cell culture medium as a single-cell suspension of epithelial tumor cells, and then the cell content is calculated by a hemocytometer, and the cell density is controlled to be 10 5 pieces / ml.

[0023] Take 80 μl and 320 μl of single-cell suspensions of concentrated silver sol and epithelial tumor cells (A431), mix them at room temperature, and transfer them to an electric shock cup sample pool. The elect...

Embodiment 2

[0025] Dissolve 12mg of silver nitrate in 50ml of deionized water, heat to 100°C to make it boil, then add 1ml of 1% trisodium citrate dropwise, stir rapidly, and keep boiling for 4 hours after the dropwise addition. After the silver sol was naturally cooled, the layers were centrifuged in a centrifuge, the supernatant was discarded, and the concentrated silver sol in the lower layer was sealed and stored at room temperature away from light for later use.

[0026] The test sample leukemia tumor cells (HL60) to be tested are first prepared with commercially available RPMI 1640 cell culture medium as a single-cell suspension of epithelial tumor cells, and then the cell content is calculated by a hemocytometer, and the cell density is controlled to be 10 6 pieces / ml.

[0027] Take concentrated silver sol and 120 μl and 480 μl of single-cell suspension of leukemia tumor cells (HL60) respectively, mix them at room temperature, and transfer them into an electric shock cup sample poo...

Embodiment 3

[0029] Dissolve 10mg of silver nitrate in 50ml of deionized water, heat to 100°C to make it boil, then add 1ml of 2% trisodium citrate dropwise, stir rapidly, and keep boiling for 6 hours after the dropwise addition. After the silver sol was naturally cooled, the layers were centrifuged in a centrifuge, the supernatant was discarded, and the concentrated silver sol in the lower layer was sealed and stored at room temperature away from light for later use.

[0030] The test sample nasopharyngeal carcinoma tumor cells (C66) to be tested are first made into a single-cell suspension of epithelial tumor cells with commercially available RPMI 1640 cell culture medium, and then the content of the cells is calculated by a hemocytometer, and the control cell density is 10 5 pieces / ml.

[0031] Take concentrated silver sol and 160 μl and 640 μl of single cell suspension of nasopharyngeal carcinoma tumor cells (C66) and mix them at room temperature, then transfer them to an electric sho...

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Abstract

The invention relates to an animal active single-cell sample treatment method wherein the animal active single-cell sample is for surface-enhanced raman spectroscopy detection. The treatment method comprises: dissolving solid silver nitrate in deionisation water and heating water to 100 DEG C., dripping sodium citrate solution into water and stirring the water until boiling, centrifuging the boiled water and taking the concentrated silver sol at the lower layer; preparing the single-cell suspension by culturing the animal active single-cell sample to be detected in the cell culture medium; performing the second centrifugation of the prepared silver sol and removing away the supernatant and adding the cell culture medium to prepare silver sol suspension and mixing the silver sol suspension with the single-cell suspension according the volume ratio of 1:4, transfering the mixture into an electric-shock cup sample cell and performing electroporation after ice bathing, and then rinsing the mixture out from the sample cell and culturing the mixture in an incubator after ice bath, finally the sample can be detected using a confocal Raman spectrometer. The treatment method has features of simpleness, quick speed, high reliability, good generality and clear and stable surface-enhanced raman spectroscopy of the inside of active cell.

Description

technical field [0001] The present invention relates to a method for pretreatment of single-cell surface-enhanced Raman spectrum detection samples, specifically a method for pretreatment of animal active cells, which are used for detection of single-cell surface-enhanced Raman spectrum detection, and using electroporation The invention discloses a method for introducing metal nanoparticles into a detection sample, belonging to the field of biomedicine. Background technique [0002] Raman spectroscopy belongs to molecular vibration spectroscopy. The number of Raman spectral lines, the size of the displacement value and the intensity of the band are all related to the vibration and rotation of the material molecule. These information reflect the conformation of the molecule and its environment. Raman spectroscopy can provide information about various normal vibration frequencies and related vibration energy levels inside the molecule, so that the functional groups present in t...

Claims

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Application Information

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IPC IPC(8): G01N21/65G01N1/28
Inventor 林居强陈荣冯尚源李永增李步洪黄祖芳蔡长美陈伟炜
Owner FUJIAN NORMAL UNIV
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