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Ratio fluorescence nano probe and application thereof

A ratiometric fluorescence, nanoprobe technology, applied in analytical chemistry and nanometer fields, can solve the problem of single color change, and achieve the effect of easy storage, small error and multi-color realization.

Inactive Publication Date: 2018-02-09
FUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, single-emission fluorescent probes usually have adverse effects from instruments, environmental conditions, and the probe molecule itself; at the same time, traditional fluorescent sensors (fluorescence quenching and fluorescence enhancement) have the problem of changing a single color

Method used

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  • Ratio fluorescence nano probe and application thereof
  • Ratio fluorescence nano probe and application thereof
  • Ratio fluorescence nano probe and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1: m -CDs@[Ru(bpy) 3 ] 2+ Construction of ratiometric fluorescent nanoprobes and their specific responses to ClO -

[0031]Firstly, 0.6 g of m-phenylenediamine was dissolved in 60 mL of absolute ethanol, and then the solution was transferred to a 100 mL polytetrafluoroethylene high-temperature reactor, and the reaction was heated at 180 °C for 12 h, and the reactor was cooled to room temperature. Afterwards, the resulting product was separated and purified by column chromatography (the eluent was dichloromethane:methanol=15:1, v / v), and the obtained solution was spin-dried with a rotary evaporator and then vacuum-dried, that is get carbon dots m - Solid crystallization of CDs. Then adjust to 20 μg / mL at a volume ratio of 6:1 m -Add 40 μg / mL Ru(bpy) to CDs 3 Cl 2 ,be made of m -CDs@[Ru(bpy) 3 ] 2+ Ratiometric fluorescent nanoprobes.

[0032] in income m -CDs@[Ru(bpy) 3 ] 2+ A series of concentrations of ClO were added to the ratiometric fluorescen...

Embodiment 2

[0033] Example 2: m -CDs@[Ru(bpy) 3 ] 2+ Preparation of agarose hydrogel and its specific response to ClO -

[0034] Towards m -CDs@[Ru(bpy) 3 ] 2+ Add agarose powder in, heat and stir m -CDs@[Ru(bpy) 3 ] 2+ The viscous gel of agarose is placed in the groove of the centrifuge tube cover, and it will form a round cake after it cools down. m -CDs@[Ru(bpy) 3 ] 2+ Agarose hydrogel.

[0035] Towards m -CDs@[Ru(bpy) 3 ] 2+ A series of concentrations of ClO were added to the agarose hydrogel - (ClO - The concentrations were 0, 10, 20, 30, 40, 50, 100, 200 μM), and after 5 min of reaction, the fluorescence intensity and color changes were observed under a 365 nm ultraviolet lamp, and a digital camera was used to photograph the fluorescence of the hydrogel imaging (eg figure 2 ).

Embodiment 3

[0036] Example 3: m -CDs@[Ru(bpy) 3 ] 2+ Ratiometric fluorescent nanoprobes for intracellular ClO - Ratio Imaging

[0037] Will m -CDs@[Ru(bpy) 3 ] 2+ Ratiometric fluorescent nanoprobes were added to culture dishes containing HeLa cells, and the 2 / 5%O 2 After culturing in an atmosphere for 1 h, the cells were washed 3-5 times with KRP buffer to remove the cells that did not enter the HeLa cells. m -CDs@[Ru(bpy) 3 ] 2+ , the cells were imaged with a confocal microscope under two fluorescent channels with excitation wavelengths of 488 nm and 543 nm; m -CDs@[Ru(bpy) 3 ] 2+ HeLa cells added 10 μM ClO - , after further incubation for 15 min, the cells were washed 3-5 times with KRP buffer to remove the ClO that did not enter the HeLa cells - , the cells were imaged with a confocal microscope under two channels of excitation wavelength of 488 nm and 543 nm, and observed m -CDs@[Ru(bpy) 3 ] 2+ Ratiometric fluorescent nanoprobes to ClO - responses (such as image 3...

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Abstract

The invention discloses a ratio fluorescence nano probe and application thereof. The characteristic that m-CDs can exclusively respond to ClO- is utilized, the m-CDs is used as a fluorescence probe, and [Ru(bpy)3]2+ is used as a reference reagent to establish the CDs@[Ru(bpy)3]2+ ratio fluorescence nano probe. After different concentrations of ClO- are added for reaction, the fluorescence color ofthe ratio fluorescence probe gradually changes from a blue-green color to a red color, so that the probe can be used for quantitative or semi-quantitative detection of the ClO-. In addition, the ratio fluorescence nano probe can be ingested into cells to achieve cell imaging, so that the probe is used for visual detection of ClO- reactive oxygen free radicals in the cells. The ratio fluorescencenano probe has the advantages of being simple in operation, wide in applicability, quick to react, capable of producing a variety of color changes and the like and can be used for multi-color, visualand online quantitative analysis of target objects.

Description

technical field [0001] The invention relates to a ratio fluorescent nanometer probe and its application, belonging to the fields of analytical chemistry and nanotechnology. Background technique [0002] Because visual detection can provide signals recognized by naked eyes, it does not require the assistance of large instruments, and is suitable for real-time and on-site detection, so it has received widespread attention. As one of the classic visualization techniques in analytical chemistry, fluorescence has the advantages of high sensitivity, high spatial and temporal resolution, and easy operation in living cells and organisms, making it widely used. However, single-emission fluorescent probes usually have adverse effects from instruments, environmental conditions, and the probe molecule itself; at the same time, traditional fluorescent sensors (fluorescence quenching and fluorescence enhancement) have the problem of changing a single color. In order to eliminate the abov...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6428G01N21/643
Inventor 郭隆华占园进邱彬林振宇陈国南
Owner FUZHOU UNIV
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