User:David Benjamin Nyer/Notebook/PcTF breast cancer/2015/10/27
PcTF breast cancer | Main project page Previous entry Next entry |
SummaryMeasured transfection efficiency of BT-474 using lipofectamine protocol, via fluorescence microscopy and flow cytometry. Measured transfection efficiency is approximately 11%. MethodsSee yesterday's page for transfection protocol. Cells were imaged using the Nikon TE inverted microscope at 100x magnification, using the 10x objective with phase contrast and GFP filter. Next, cells were washed with phosphate buffered saline, trypsinized and resuspended in 0.5 mL PBS, then measured for FSC, SSC, and GRN fluorescence on the C6 flow cytometer. ResultsFluorescence microscopy: Flow cytometry: Well 1 - Count vs. GRN fluorescence Well 2 - Count vs. GRN fluorescence Well 3 (neg) - Count vs. GRN fluorescence Conclusion & Next StepsTransfection efficiency for pMaxGFP in BT-474 cell line is ~11%. Next steps are to perform optimization of Neon transfection system using BT-474 cell line. |