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Preparation and application of fluorescent carbon dot nanoprobe for detecting methylene blue and living cell imaging

A methylene blue and nano-probe technology, applied in the field of biochemical analysis, can solve the problems of strict reaction conditions, expensive and difficult to obtain raw materials, complicated process, etc., and achieve the effects of wide linear response range, stable imaging, and simple preparation

Inactive Publication Date: 2015-04-22
GUANGXI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In these methods, some raw materials are expensive and difficult to obtain, some processes are more complicated, some reaction conditions are relatively strict, and energy consumption is relatively large.

Method used

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  • Preparation and application of fluorescent carbon dot nanoprobe for detecting methylene blue and living cell imaging
  • Preparation and application of fluorescent carbon dot nanoprobe for detecting methylene blue and living cell imaging
  • Preparation and application of fluorescent carbon dot nanoprobe for detecting methylene blue and living cell imaging

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Preparation of fluorescent nanomaterial CDs

[0032] Fluorescent nanomaterial carbon dots (CDs) were prepared by pyrolyzing lychee kernels, as follows: weigh 5.0 g of lychee kernels in a crucible, heat them to 300 °C in a muffle furnace, take them out after 2 hours, and grind them with a mortar. Then add 20 mL of ultrapure water, sonicate for 30 min, filter, centrifuge at 10,000 rpm for 10 min, and store the supernatant at 4°C in the dark.

Embodiment 2

[0034] Utilize methylene blue to measure the quenching action of CDs fluorescent probe in the determination of methylene blue in water, specifically:

[0035] In the PB buffer solution with pH=8.0, in the presence of 50 μmol / L methylene blue solution, methylene blue has a very fast response to CDs. At the excitation wavelength of 320 nm, the fluorescence of CDs is quickly quenched. When , the fluorescence is quenched by about 80% ( image 3 ), and the fluorescence remained essentially unchanged in the ensuing 1 hour. The results indicated that methylene blue quenched the fluorescence of CDs very much and quickly, implying that the fluorescent probe was fast and stable. The fluorescent probe detects methylene blue with a detection limit as low as 0.05 μmol / L, and a linear response range: 0.8-10 μmol / L ( R 2 = 0.9930) ( Figure 4 ).

Embodiment 3

[0037] Application of CDs Fluorescent Probes in Live Cell Imaging

[0038] Add 80 μg / ml of the fluorescent CDs to a cell culture dish and incubate with living cells for 24 hours, wash the cells with PBS to remove CDs that have not entered the cells, and observe with a laser confocal microscope system. Under excitation at 405 nm, the cells continue to emit blue color fluorescence.

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Abstract

The invention relates to preparation and application of a fluorescent carbon dot nanoprobe for detecting methylene blue and living cell imaging. Under the excitation wavelength of 320nm, the maximum emission wavelength of carbon dots (CDs) appears at 440nm and strong blue fluorescence is emitted. Methylene blue can be used for quenching the fluorescence of the carbon dots, and in a cell culture dish, a living cell incubates for 24 hours with the methylene blue, and the cell is observed by using a laser confocal microscopy system and continuously emits blue fluorescence. According to the preparation and the application, the carbon dots carbonized by semen litchi can be used for detecting the methylene blue by quenching a fluorescent signal. As an effective means for detecting the methylene blue, the method shows a remarkable advantage. The detection limit is as low as 0.05micro mol / L and the linear response range is 0.8-10micro mol / L [R<2>=0.9930]. As being a fluorescent probe, a sensing system has the optical characteristics of simplicity, low cost, greenness, high selectivity, rapidness and sensitivity and is successfully applied to methylene blue detecting and living cell imaging analysis. The sensing method has a broad application prospect in the field of environmental analysis and the cell imaging analysis.

Description

technical field [0001] The invention belongs to the field of biochemical analysis, in particular to the preparation and application of a fluorescent carbon dot nanoprobe for detecting methylene blue and live cell imaging. Background technique [0002] Carbon dots (CDs), as a new member of the carbon nanomaterial family, are carbon nanoparticles with a diameter less than 10 nm, usually composed of SP 2 It is composed of hybridized carbon atoms, oxygen- and hydrogen-rich substances. As a new type of fluorescent carbon nanomaterial, carbon dots have special fluorescent properties, such as adjustable excitation and emission wavelengths, good photostability, and no light flickering phenomenon. Compared with traditional organic fluorescent dyes and metal quantum dots, CDs, It has many advantages, and its unique luminescent properties and biocompatibility have good application prospects in the fields of photocatalysis, luminescent devices, optoelectronics, biochemical analysis, ce...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64B82Y5/00B82Y15/00
Inventor 赵书林薛茗月湛志华张亮亮
Owner GUANGXI NORMAL UNIV
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