New use of cyanine dyes
A technology of use and buffer solution, applied in the field of supramolecular probes, can solve the problems of increased detection cost, detection difficulty, low sensitivity and accuracy, etc., and achieve the effect of short detection time and simple operation.
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Embodiment 1
[0038] Fifty nude mice were reared, 20 were used as a blank control, and 30 were inoculated with lung cancer cells (A549) in nude mice. After the tumor grew, the nude mice inoculated with A549 were taken from tumor tissues, and the blank control group was directly taken from lung cells to make The single cell suspension was washed and suspended with pH 6 sodium phosphate-sodium hydrogen phosphate buffer to obtain 20 normal cell suspension samples and 30 tumor cell suspension samples. The concentration of each cell suspension sample was 10 4 ~10 9 The range of cells / mL.
[0039] Add the compound of the following formula to each sample, so that the final concentration of the compound in each sample is 10μM
[0040]
[0041] Each sample was incubated in a 37°C incubator for 1 hour, and then incubated in a 4°C refrigerator for 1 hour.
[0042] After the incubation, the sample was centrifuged to remove the dye solution, and the cells were washed with pH 6 sodium phosphate-sodium hydrogen ...
Embodiment 2
[0047] The same procedure as in Example 1 was used to detect tumor cells and normal lung cells of the above 50 nude mice. The difference is that the compounds used are:
[0048]
[0049] The final concentration of the compound in each sample is 50μM, the incubation temperature and time are 15 minutes at 37°C, and 15 minutes at 4°C. The fluorescence signal of each sample above 670nm is detected by flow cytometry to obtain the staining rate of each sample. , Get the dyeing rate of each sample.
[0050] Test result: The staining rate of all lung cancer samples is greater than 75%, and the staining rate of all normal lung cell samples is less than 15%. Therefore, the diagnosis method of this embodiment can easily distinguish normal lung cells from lung tumor cells.
Embodiment 3
[0052] The same procedure as in Example 1 was used to detect tumor cells and normal lung cells of the above 50 nude mice. The difference is that the compounds used are:
[0053]
[0054] The final concentration of the compound in each sample is: 150μM, the incubation temperature and time are 8 hours at 37°C, and 1 hour at 4°C. The fluorescence signal of each sample above 670nm is detected by flow cytometry, and the staining rate of each sample is obtained. .
[0055] Test result: the staining rate of all lung cancer samples is greater than 85%, and the staining rate of all normal lung cell samples is less than 20%. Therefore, the diagnosis method of this embodiment can easily distinguish normal lung cells from lung tumor cells.
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