Turn-on type fluorescent molecular probe responding to biological cell viscosity as well as preparation method and application of Turn-on type fluorescent molecular probe
A technology of fluorescent molecular probes and biological cells, which is applied in the field of fluorescent molecular probe detection, can solve the problems that viscometers cannot use living cells, etc., and achieve the effects of obvious changes in ultraviolet color, little harm, and sensitive response
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Embodiment 1
[0050] Such as Figure 1-4 As shown, a Turn-on fluorescent molecular probe responsive to the viscosity of biological cells, 2-2-2-chloro-3-(1,3-dihydro-3,3-dimethyl-1-propyl -2-hydro-indol-2-ylidene)ethylidene-1-cyclohexen-1-ylethenyl-3,3-dimethyl-1-propylindolium chloride and tetrazolium Be reaction raw material, obtain 2-[2-(2-chlorocyclohexanone)]-3,3-dimethyl-1-proguanil hydrochloride-3-hydrogen-indole is fluorescent group, and this molecular probe is 2-[2-(2,3,4,5,6,7,8,9,13,14-decahydro)]-12hydro-1,10-furan-(1,4,7,10-tetra Azole)-15-vinyl-3,3-dimethyl-n-propyl-3 hydrogen-indole, its chemical structural formula is:
[0051]
[0052] The present invention also includes a method for preparing a Turn-on fluorescent molecular probe responsive to biological cell viscosity, the fluorescent molecular probe has the above-mentioned structure, comprising the following steps:
[0053] 2-2-2-Chloro-3-(1,3-dihydro-3,3-dimethyl-1-propyl-2-hydro-indole-2-ylidene)ethylene-1- Cyclo...
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 them evenly in the test tube, and then add 10 μL of Molecular Probe F with a concentration of 1 mM 474 DMSO solution, mix evenly, and complete the sample measurement in the fluorescence cuvette;
[0062] (3) Test fluorescence intensity by fluorescence spectrophotometer, obtain fluorescence intensity ratio, detect the excitation wavelength of the fluorescence intensity of 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 fluorescence inten...
Embodiment 3
[0065] Detection Molecular Probe F 474 The UV absorption steps at minimum and maximum viscosity include,
[0066] Take 2 parts of the methanol and glycerol ratio: 10:0 and 0:10 solutions, mix them evenly in the test tube, and then add 10 μL of 1mM F 474 DMSO solution, mixed uniformly, finished sample measurement in cuvette, obtained the ultraviolet absorption spectra of two samples. Figure 7 is Molecular Probe F 474 UV absorption plots at maximum and minimum viscosities, where the minimum viscosity is methanol and the maximum viscosity is glycerol.
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