Method for improving cell calcium flow fluorescence detection
A fluorescence detection and cell technology, which is applied in the field of fluorescence detection to improve the calcium flow of cells, can solve the problems of high background signal, low fluorescence signal, and affecting the judgment of results, and achieve the effect of reducing background signal, simple operation process, and enhancing calcium flow signal
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Embodiment 1
[0029] Embodiment 1 (traditional method)
[0030] Step 1: Prepare the cell plate, sow HEK-293-D2 and HEK-293-Mu in a 96-well plate at a density of 80,000 cells per well (the type of the well plate is black around the bottom, pre-coated with Poly-D- lysine), cultivated for 16~24 hours;
[0031] Step 2: For high-throughput real-time fluorescence detection and analysis, add 100 μL of fluorescent dye solution (Calcium-6 solution) to each well and place it in an incubator at 37°C for 1 hour to perform high-throughput real-time fluorescence detection and analysis.
[0032] The analysis is shown in Figure 2(A) and Figure 2(B), in which 1A-3H is the repeated measurement of HEK-293-D2 cells, 1A-3A is the blank control, and 1B to 3H are dopamine with decreasing concentrations, namely 10 μM, 2 μM, 400 nM, 80 nM, 16 nM, 3.2nM, 0.64 nM, 0.0128 nM; 4A-6H are HEK-293-Mu replicate wells, 4A to 6G are endorphins with decreasing concentrations, That is, 10 μM, 2 μM, 400 nM, 80 nM, 16 nM, 3.2 ...
Embodiment 2
[0034] Embodiment 2 (the inventive method)
[0035] Step 1: Prepare the cell plate, sow HEK-293-Mu cells and HEK-293-D2 cells in a 96-well plate at a density of 80,000 cells per well (the type of the well plate is black around the bottom, pre-coated with Poly -D-lysine), cultivated for 20 hours;
[0036] Step 2: prepare amaranth-containing buffer solution, take amaranth powder and dissolve it in high-throughput real-time fluorescence detection buffer solution to obtain amaranth-containing buffer solution with a concentration of 0.5 mg / mL;
[0037] Step 3: High-throughput real-time fluorescence detection and analysis, add 100 μL of fluorescent dye solution to each well of the 96-well plate, put it in an incubator at 37°C for 1 hour, and then detect the fluorescence in the 96-well plate The dye solution was blotted dry, and 100 μL of 0.5 mg / mL amaranth-containing buffer was added to each well for high-throughput real-time fluorescence detection analysis.
[0038] The analysis ...
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