Method for marking eukaryotic cells by taking coptisine as fluorescent probe and application of coptisine
A fluorescent probe, coptisine technology, applied in the field of biomedicine, can solve the problems of not being able to label with living cells well, poor stability, short reagent storage time, etc., and achieves easy preparation and test repeatability, short time-consuming, operation simple effect
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0051] Fluorescence spectrum of coptisine
[0052] The structure of the yellow-sized alkaline is as follows:
[0053] The tubilate was dissolved with DMSO (Sigma), and the mother liquor concentration was 20 mm / L. 2 μL of mother liquor was taken to 2 ml DMSO to make the yellow alkaline final concentration of 20 μm / L. A new 96-well plate was taken, and 200 μl of diluted tinoids were added to each well; DMSO was used as a blank solvent control. Three complex holes are set up in each group, and the multi-function enzyme labeled instrument is removed. First fixing the emission light wavelength is 550 nm, and the excitation spectrum of the yellow alkali is scanned; then the fixed excitation wavelength is 460 nm, and the scan is obtained to obtain the emission spectrum of the tubilator, such as Figure 1-2 . by Figure 1-2 It can be seen that there are two peaks in the hunal alkali excitation spectrum, and the maximum excitation wavelength of green fluorescence is 460 nm, and the maxim...
Embodiment 2
[0055] The hela cells were placed into the vessel (35mm), and three complex holes were set each set. The cells were removed from the incubator, discard the medium, and wash it once with PBS. Then, a medium obtained by the tuberi (DMEM (GIBCO) (containing 10% FBS)) was diluted to 20 μm / L, and 1.5 ml of medium diluted tantaline (reversed dosing, such as). First incubate 8 h group, incubated 6 h in 2 h, push it in this type), place a cell incubator (37 ° C, 5% CO) 2 The corresponding time was incubated, and the control was incubated with a volume of DMSO with a volume of the xylullette of the experimental group. The cells were taken out from the incubator, discard the medium, and washed 2 times with PBS. 500 μL of 0.25% truncazone (containing EDTA) was added to 2-5 min. Then, 500 μl of DMEM (containing 10% FBS) was added to the dish, and the cells were collected into the centrifugal tube to centrifuge at 500 rpm. Pour the supernatant, the cells were resuspended with PBS and centrif...
Embodiment 3
[0057] Different concentrations of tabtisine labeled cell fluorescence intensity
[0058] Wrap HELA cells to 6-well plates, each set of three complexes. The cells were removed from the incubator, discard the medium, and wash it once with PBS. After the tuberi is diluted with medium (DMEM (containing 10% FBS)) to 20 μm / L, 10 μm / L, 5 μm / L, 1 μm / L, and 1.5 ml of medium diluted noodles per well. Base, place cell incubator (37 ° C, 5% CO 2 Incubate 6 h, the control was incubated with a volume of DMSO with a volume of DMSO with a volume of the experimental group. The cells were taken out from the incubator, discard the medium, and washed 2 times with PBS. To give a 500 μl of 0.25% trunca (containing EDTA) digestion 2-5 min. A 500 μl of DMEM (containing 10% FBS) was then added to each well, and the cells were collected into the centrifugal tube to centrifugally 500 rpm. Pour the supernatant, the cells were resuspended with PBS and centrifuged at 500 rpm for 5 min. After centrifug...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More