Team:Heidelberg/Project/Measurement Standard
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==Methods== | ==Methods== | ||
- | To measure GFP and BFP fluorescence, we used microscopy and | + | To measure GFP and BFP fluorescence intensity, we used microscopy and flow cytometer. Fluorescence was fist evaluated using the Leica DM IRB epifluorescence microscope. Only cells which were positive for transfection were measured. |
First, the cells were washed with 1x PBS and detached from the plate using Trypsin. 30µl Trypsin was added to each well, incubated for ten minutes at room temperature. Cells were resuspended in 170µl PBS/BSA and replicates for each condition were pooled into 24 well plates. 200µl from each well were used for FACS measurements, 100-150µl were used for confocal microscopy. | First, the cells were washed with 1x PBS and detached from the plate using Trypsin. 30µl Trypsin was added to each well, incubated for ten minutes at room temperature. Cells were resuspended in 170µl PBS/BSA and replicates for each condition were pooled into 24 well plates. 200µl from each well were used for FACS measurements, 100-150µl were used for confocal microscopy. | ||
Revision as of 23:51, 23 October 2010
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