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Method for detecting trace amounts of Ca<2+>, Sr<2+> and Ba<2+> in active cancer cells through single-channel fluorescence imaging

A fluorescence imaging, single-channel technology, used in measurement devices, fluorescence/phosphorescence, material analysis by optical means, etc., can solve problems such as poor selectivity, inconvenient detection, and inability to achieve efficient detection, and achieve convenient monitoring and high sensitivity. , the effect of less interference

Active Publication Date: 2018-09-18
GUIZHOU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] There have been many reports as probes for the detection of trace metal ions in cell imaging, but most of them are single-probe detection of a single target ion, which is inconvenient for detection, poor in selectivity, and cannot achieve efficient detection of multiple target ions in cells by a single probe. question

Method used

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  • Method for detecting trace amounts of Ca&lt;2+&gt;, Sr&lt;2+&gt; and Ba&lt;2+&gt; in active cancer cells through single-channel fluorescence imaging
  • Method for detecting trace amounts of Ca&lt;2+&gt;, Sr&lt;2+&gt; and Ba&lt;2+&gt; in active cancer cells through single-channel fluorescence imaging
  • Method for detecting trace amounts of Ca&lt;2+&gt;, Sr&lt;2+&gt; and Ba&lt;2+&gt; in active cancer cells through single-channel fluorescence imaging

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] 1. Probe preparation:

[0055] The chemical structural formula of the probe is:

[0056]

[0057] Its synthetic route is as follows:

[0058]

[0059] The specific preparation method is:

[0060] ① Preparation of Intermediate 1:

[0061] Take 100 grams (0.67mol) of p-tert-butylphenol, add it to a 1000ml three-necked round-bottomed flask, then add 1.55g (33.75mmol) sodium hydroxide and 80ml of 37% formaldehyde solution in sequence, and blow out more than 90% of it under nitrogen flow Most of the water, heated to 115°C (110-120°C), mechanically stirred for 2h, stopped the reaction, cooled to room temperature, poured 500ml of 85°C diphenyl ether into it, and continued to stir, the mixture in the flask Heating to reflux temperature, after reacting for 3 hours, stop the reaction, cool to room temperature, add 2000ml ethyl acetate into the flask, stir until white precipitate precipitates out, let stand, suction filter, and successively use 50ml ethyl acetate, 50ml ac...

Embodiment 2

[0070] Embodiment 2. The preparation of reagent:

[0071] (1) Preparation of probe solution: Weigh 10.3 mg of probe (prepared by the above method), dissolve it in N,N-dimethylformamide, and prepare 10 mL of a solution with a concentration of 1 mM.

[0072] (2)Ca 2+ Preparation of ion stock solution: Weigh 0.3110 g of calcium perchlorate, dissolve it in ultrapure water, prepare 50 mL of a solution with a concentration of 20 mM, and then dilute to the required concentration with ultrapure water.

[0073] (3)Sr 2+ Preparation of ion stock solution: Weigh 0.3946g of strontium perchlorate, dissolve it in ultrapure water, prepare 50mL of solutions with a concentration of 20mM, and then dilute to the required concentration with ultrapure water.

[0074] (4) Ba 2+ Preparation of ion stock solution: Weigh 0.3903g of barium perchlorate, dissolve it in ultrapure water, prepare 50mL of solutions with a concentration of 20mM, and then dilute to the required concentration with ultrapure ...

Embodiment 3

[0082] Embodiment 3: Fluorescence spectrometry detects Ca 2+ 、Sr 2+ 、Ba 2+

[0083] Add the probe (1mM, 100μL) into a 10mL volumetric flask, dilute it with N,N-dimethylformamide / water, so that the composition of the probe solution is N,N-dimethylformamide with a volume ratio of 9 / 1 Amide / water solution, shake well. Add about 3ml into a 1cm cuvette, and measure the fluorescence spectrum with 405nm as the fluorescence excitation wavelength.

[0084] In N,N-dimethylformamide / water solution with a volume ratio of 9 / 1, the probe solution with a concentration of 10 μM has fluorescence emission at a wavelength of 510 nm. Add 200 μM metal ion Li + , Na + , K + , Mg 2+ , Ca 2+ , Ba 2+ , Hg 2+ ,Sr 2+ , Zn 2+ , Cd 2+ , Ni 2+ ,Co 2+ , Pb 2+ , Fe 3+ , Cr 3+ , Ag 2+ , Cu 2+ , no obvious change in the fluorescence spectrum was observed, only the addition of 200 μM Ca 2+ 、Sr 2+ 、Ba 2+ Significantly reduce the fluorescence peak of the probe at 510nm (see figure 1 ).

...

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Abstract

The invention discloses a method for detecting trace amounts of Ca<2+>, Sr<2+> and Ba<2+> in active cancer cells through single-channel fluorescence imaging. By using classic calixarene [4] derivativeas a probe, and the trace amounts of Ca<2+>, Sr<2+> and Ba<2+> in the active cancer cells are respectively detected through the single-channel fluorescence imaging. According to the method disclosedby the invention, detection on multi-target ions can be realized; a coordination compound of a probe-Ca<2+>, a probe-Sr<2+> and a probe-Ba<2+> can be respectively formed by the probe disclosed by theinvention and Ca<2+>, Sr<2+> and Ba<2+> in the cells, and intense fluorescence with different wavelengths can be emitted, so that detection is conveniently carried out, the visibility is good, the sensitivity is good, the selectivity is good, and real-time on-line detection of the Ca<2+>, Sr<2+> and Ba<2+> in the cells can be realized.

Description

technical field [0001] The invention relates to a single-channel fluorescence imaging method for detecting trace ions in living cells, in particular to a single-channel fluorescence imaging method for detecting trace amounts of Ca in active cancer cells 2+ 、Sr 2+ and Ba 2+ Methods. Background technique [0002] In cell biology and medical research, fluorescent probes are the most powerful tools for allowing researchers to monitor events in living cells and living organisms in real time due to their high spatial resolution and minimal disturbance of the living body. One of them is being paid more and more attention to and used. Probes are particularly important for the study of metal ion transport carriers and the crossing behavior of cell membranes in drug delivery systems. Fluorescence imaging to detect intracellular ions can provide the most direct information on the temporal and spatial distribution of ions in living systems. Due to its non-invasive, high sensitivity,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6428G01N21/6486G01N2021/6432
Inventor 曾晞廖贤方浚安许万里田雁牟兰
Owner GUIZHOU UNIV
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