Don’t judge; I’m green, but normal!
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One of the research projects I am currently working on requires the use of normal primary mammary epithelial cells to study the molecular mechanism of the transformation of human breast cancer. In this project we are investigating whether overexpression of splicing factors in normal tissues will actually cause malignant transformation. To determine whether ectopic overexpression of splicing factor results in immortalization and/or transformation, I transduced normal breast epithelial cells with lentiviral particles carrying GFP tagged cDNA expression cassettes. I used an empty expression vector that only carries the enhanced green fluorescent protein(e-GFP) as a negative control. The availability of GFP and its derivatives has thoroughly redefined fluorescence microscopy and it is one of the wildly used methods in cell biology and other biological disciplines. Morevever the GFP is much less harmful when expressed in living cells compared to other fluorescent molecules such as FITC . In this project, the GFP allows me to monitor our lentivirus transduced normal primary mammary epithelial cells in real time. My micrograph features normal primary mammary epithelial cells cultivated on tissue culture plates that have been transduced with lentiviral particles carrying GFP expression cassette. Two images, phased-contrast and fluorescence, were acquired from an inverted fluorescence microscope. Later, phased-contrast micrograph overlaid with the fluorescence micrograph to obtain this final image.