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Method for simultaneous fluorescence imaging of As < 3 + >, Pb < 2 + > and Hg < 2 + > in cells

A fluorescence imaging, cell technology, applied in fluorescence/phosphorescence, material analysis by optical means, instruments, etc., can solve the problems that have not been reported in the literature, and achieve the effect of enhanced sensitivity and high selectivity detection

Pending Publication Date: 2021-08-24
QINGDAO UNIV OF SCI & TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0010] So far, hollow porous nanomaterials have been used to construct nanocapsule-nucleic acid biomolecules complexes for intracellular As 3+ , Hg 2+ and Pb 2+ Simultaneous fluorescence imaging technology has not been reported in the literature

Method used

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  • Method for simultaneous fluorescence imaging of As &lt; 3 + &gt;, Pb &lt; 2 + &gt; and Hg &lt; 2 + &gt; in cells

Examples

Experimental program
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Effect test

Embodiment 1

[0036] An available intracellular As 3+ , Pb 2+ and Hg 2+ A method for preparing a nanocapsule-nucleic acid biomolecule complex for simultaneous fluorescence imaging, comprising the steps of:

[0037] (1) Separately screen, design and synthesize three kinds of nucleic acid biorecognition molecules to ensure that they are respectively and only for the target metal ion As 3+ , Pb 2+ and Hg 2+ Have a specific response, but no response to any other coexisting ions and interference ions;

[0038] (2) Add 200 μL of 11.664 mg / mL polydiallyldimethylammonium chloride solution to 400 μL of nano-gold carrier solution with a hollow and porous structure, centrifuge at 37 ° C for 10 h, and use Tris- Washing with HCl buffer solution;

[0039] (3) Add 2 μL of fluorescent molecule Cy5 solution (final concentration 1.0×10 -5 mol / L), diluted to 100 μL with Tris-HCl buffer solution with pH = 7.4, and 10 μL of identifiable As was added after 10 h at 37 °C 3+ Nucleic acid biomolecule soluti...

Embodiment 2

[0048] an intracellular As 3+ , Pb 2+ and Hg 2+ A method for simultaneous fluorescence imaging comprising the following steps:

[0049] (1) Take an appropriate amount of cell suspension, centrifuge, remove the supernatant, add 300 μL containing As 3+ , Pb 2+ and Hg 2+ The final concentration was 1.0×10 -11 Cultivate the cells with M solution, centrifuge at 37°C for 20.0min, and remove the supernatant;

[0050] (2) Dilute the nanocapsule-nucleic acid biomolecule complex prepared in Example 1 with a Tris-HCl buffer solution of pH=7.4, add it to the cells treated in the previous step, and incubate in a constant temperature water bath at 37°C Cell 15.0min;

[0051] (3) Place the incubated cells on the stage of a confocal microscope, and use excitation wavelengths of 649nm, 559nm, and 488nm to detect intracellular As 3+ , Pb 2+ and Hg 2+ laser confocal scanning imaging;

[0052] Wherein, the cells are Hela cells.

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Abstract

Environmental and water pollution caused by high-toxicity heavy metals is always a serious problem faced by human beings. As < 3 + >, Pb < 2 + > and Hg < 2 + > are environmental pollutants which have serious toxicity and are widely distributed, and can seriously threaten the environment and human health even at low concentration. The invention provides a method for simultaneous fluorescence imaging of As < 3 + >, Pb < 2 + > and Hg < 2 + > in cells by using a nanocapsule-nucleic acid biomolecule compound. The provided and constructed nanocapsule-nucleic acid biomolecule compound has the advantages of being simple in structure, good in biocompatibility, ingenious in design, easy to prepare, stable in performance, high in cell membrane infiltration capacity, high in intracellular release controllability, high in specificity, short in response time and the like and high-sensitivity and high-selectivity simultaneous fluorescence imaging of As < 3 + >, Pb < 2 + > and Hg < 2 + > in cells can be conveniently and quickly realized.

Description

technical field [0001] The present invention relates to As 3+ , Pb 2+ , Hg 2+ Simultaneous detection and imaging of , specifically related to a method for intracellular As 3+ , Pb 2+ and Hg 2+ The simultaneous fluorescence imaging of the invention belongs to the field of environmental monitoring and nanobiotechnology. Background technique [0002] Environmental and water pollution caused by highly toxic heavy metals has always been a major concern of people. Highly toxic heavy metals can enter the food chain through air, water and soil and seriously endanger the ecosystem. [0003] Arsenic is an element that exists widely in nature and has metal-like properties. Arsenic and its compounds have been recognized as carcinogens by many international authorities such as the International Agency for Research on Cancer under the World Health Organization (WHO) and the US Environmental Protection Agency (EPA). Arsenic pollution mainly comes from the discharge of three wastes f...

Claims

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Application Information

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IPC IPC(8): G01N21/64G01N33/53
CPCG01N21/6486G01N33/5308
Inventor 王卫侯晓珊王世颖万均宋志灵罗细亮
Owner QINGDAO UNIV OF SCI & TECH
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