Simultaneous fluorescence imaging method for Hg2+ and Ag+ in cells

A fluorescence imaging and cell technology, which is applied to nanocapsule-nucleic acid biomolecule complexes and their preparation. At the same time in the field of imaging, it can solve the problems that have not been reported in the literature, and achieve the effects of wide application range, simple structure and improved detection sensitivity.

Pending Publication Date: 2021-08-13
QINGDAO UNIV OF SCI & TECH
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  • Description
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Problems solved by technology

So far, nanocapsule-nucleic acid biomolecular complexes have been constructed for intracellular Hg by using hollow porous nanomaterials and utilizing the biorecognition effect of base mismatches on two metal ions. 2+ and Ag + Simultaneous imaging technology has not been reported in the literature

Method used

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  • Simultaneous fluorescence imaging method for Hg2+ and Ag+ in cells

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Experimental program
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Effect test

Embodiment 1

[0030] One available for intracellular Hg 2+ and Ag + A method for preparing a nanocapsule-nucleic acid biomolecule complex for simultaneous fluorescence imaging, comprising the steps of:

[0031] (1) By selecting different types and numbers of bases, two kinds of nucleic acid biorecognition molecules were designed and synthesized to ensure that they are respectively and only for the target metal ion Hg 2+ or Ag + Have a specific response, but no response to any other coexisting ions and interference ions;

[0032] (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 buffer with MOPS with pH = 7.0 solution cleaning;

[0033] (3) Add 2 μL of rhodamine B solution (final concentration 1.0×10 -5 mol / L), diluted to 100 μL with MOPS buffer solution with pH=7.0, and 10 μL of identifiable Hg was added after 10 h at 37 °C 2+ Nucleic acid biomolecu...

Embodiment 2

[0040] an intracellular Hg 2+ and Ag + A method for simultaneous fluorescence imaging, comprising the steps of:

[0041] (1) Take an appropriate amount of cell suspension, centrifuge, remove the supernatant, add 100 μL containing Hg 2+ and Ag + The concentration is 1.0×10 -12 Cultivate the cells with M solution, centrifuge at 37°C for 20.0min, and remove the supernatant;

[0042] (2) Dilute the nanocapsule-nucleic acid biomolecule complex prepared in Example 1 with MOPS buffer solution, add it to the cells treated in the previous step, and incubate the cells in a constant temperature water bath at 37°C for 15.0min;

[0043] (3) Place the incubated cells on the stage of a confocal microscope, and use excitation wavelengths of 535nm and 648nm to detect intracellular Hg 2+ and Ag + laser confocal scanning imaging;

[0044] Wherein, the cells are Hela cells.

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Abstract

Hg2+ and Ag+ are two environmental pollutants which have serious toxicity and are widely distributed, the Hg2+ and the Ag+ can seriously threaten the environment and human beings even under low concentration, and serious and permanent damage is caused. The invention provides a simultaneous fluorescence imaging method for Hg2+ and Ag+ in cells. The simultaneous fluorescence imaging method can sensitively and efficiently realize simultaneous fluorescence imaging for Hg2+ and Ag+ in cells by adopting a nanocapsule-nucleic acid biomolecule compound. The nanocapsule-nucleic acid biomolecule compound provided and constructed by the invention has the advantages of being simple in structure, good in biocompatibility, ingenious in design, simple to prepare, stable in performance, high in cell membrane infiltration capacity, high in intracellular release controllability, high in specificity to Hg2+ and Ag+, short in response time, convenient to monitor in real time and the like, and can realize high-sensitivity and high-selectivity simultaneous fluorescence imaging of Hg2+ and Ag+ in cells conveniently and quickly.

Description

technical field [0001] The present invention relates to Hg 2+ and Ag + Simultaneous imaging, specifically involving a method for intracellular Hg 2+ and Ag + The nanocapsule-nucleic acid biomolecule complex for simultaneous fluorescence imaging and its preparation method belong to the field of environmental monitoring and nanobiotechnology. Background technique [0002] Mercury ion (Hg 2+ ) and silver ions (Ag + ) are two metal ions with severe toxicity and are also widely distributed environmental pollutants, which pose serious threats to the environment and human beings even at low concentrations, causing severe and permanent damage. More seriously, through polluted water bodies, Hg 2+ and Ag + It can continuously accumulate in agricultural and aquatic products and enter the human food chain. If prolonged exposure to Hg 2+ and Ag + In a polluted environment, it will lead to the slow occurrence of degenerative diseases of the human body and nervous system. For th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64G01N33/53
CPCG01N21/6486G01N33/5308
Inventor 王卫唐凯王世颖李欣万均罗细亮
Owner QINGDAO UNIV OF SCI & TECH
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