Field of research: Cell Biology (Biophotonics)
Lab: Diana Bratu
School: Manhattan Hunter Science High School
Year: 10th/11th Grade
Over the summer, I worked in Professor Diana Bratu’s lab. I focused on the visualization of RNAs and proteins within the Drosophila melanogaster (fruit fly) oocyte. During my internship, I performed fluorescence in situ hybridization (FISH) to detect my target mRNA (oskar) or immuno-fluorescence (IF) to stain for various proteins (Staufen, Actin) which I observed using a spinning disc confocal microscope. The experiments were performed using wild type flies. I used Phalloidin-FITC to stain for Actin and visualize the developmental stages of the oocytes. I used an anti-Staufen antibody to stain for Staufen in combination with an AlexaFluor555 secondary antibody. I used molecular beacon probes labeled with Cy5 to specifically detect oskar mRNA. I compared these results with staining for Staufen and oskar mRNA in a fly expressing a protein tagged with green fluorescent protein (GFP) that binds to oskar mRNA. My goal was to determine whether the FISH and IF protocols can be combined to simultaneously study the localization of mRNAs and proteins in fixed cells. Also I compared the localization of mRNAs using the signal generated by GFP and that generated by molecular beacon probes that detect the same oskar mRNA molecules.