A turn-on fluorescent molecular probe responsive to the viscosity of biological cells and its preparation method and application
A fluorescent molecular probe and biological cell technology, applied in the field of fluorescent molecular probe detection, can solve the problem that a viscometer cannot use living cells, etc., and achieve the effects of low detection limit, obvious changes, and little harm to biological cells.
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Embodiment 1
[0050] Such as Figure 1-4 As shown, a Turn-on fluorescent molecular probe that responds to the viscosity of biological cells, As the reaction raw material, molecular probe F was obtained 474 , its chemical structure is:
[0051]
[0052] The present invention also includes a method for preparing a turn-on fluorescent molecular probe responsive to the viscosity of biological cells, the fluorescent molecular probe having the above-mentioned structure, comprising the following steps:
[0053] Will added to the flask, and Finally, chloroform was added as a solvent, and the reaction was carried out at 80°C. After the reaction was completed, the solution was cooled to room temperature and separated by column chromatography to obtain the product, which was named as F 474 , is a dark red solid.
[0054] Among them, the and The molar ratio is 1:5;
[0055] The reaction time is 12h.
[0056] figure 1 is Molecular Probe F 474 synthetic route map; figure 2 is Molecul...
Embodiment 2
[0059] Build Molecular Probe F 474 The titration linear curve steps against viscosity include,
[0060] (1) Preparation of pH = 7.40, concentration of 20mM PBS buffered saline solution; concentration of 1mM Molecular Probe F 474 DMSO solution;
[0061] (2) The ratios of methanol and glycerol are: 10:0, 8:2, 7:3, 6:4, 5:5, 4:6, 3:7, 2:8, 1:9 and 0 :10 solution in 10 parts in total, mix evenly in the test tube, and then add 10 μL of Molecular Probe F with a concentration of 1mM 474 DMSO solution, mix evenly, and complete the sample measurement in the fluorescence cuvette;
[0062] (3) Test the fluorescence intensity by a fluorescence spectrophotometer to obtain a fluorescence intensity ratio, and the excitation wavelength of the fluorescence intensity for detecting cell viscosity is 400nm;
[0063] (4) take the content of glycerol as the abscissa respectively, and take the fluorescence intensity ratio as the ordinate to obtain a linear equation about viscosity and the fluore...
Embodiment 3
[0065] Detection Molecular Probe F 474 The UV absorption steps at minimum and maximum viscosity include,
[0066] Take 2 parts of methanol and glycerol at the ratio of 10:0 and 0:10, mix them evenly in the test tube, and then add 10 μL of 1 mM F 474 DMSO solution, mixed evenly, completed the sample measurement in a cuvette, and obtained the ultraviolet absorption spectra of two samples. Figure 7 is Molecular Probe F 474 The UV absorption graph in the maximum and minimum viscosities, where the minimum viscosity is methanol and the maximum viscosity is glycerol.
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