Final Weeks at the Phillips

My last two weeks at the Phillips Collection were filled with plenty of exciting projects and meetings across several departments. While I was sad to leave the my internship at the Phillips last Friday, I left with the sense that I made some valuable contributions over the past ten weeks. I also left with the sense that the relationships that I built over the course of this summer will not fade quickly, and I look forward to returning to the Phillips as often as possible with friends and family to share all that the museum has to offer.

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Live Cell Imaging of Drosophila Gonads (Update 3)

The end of the summer session is fast approaching and so much has happened since my last post.

First of all, I was finally able to image some differentiating gonads! I found that adding 450 microliters of freshly aliquoted Atlanta Biological’s heat inactive fetal bovine serum (FBS) to my live cell imaging culture media supports cell differentiation. However, I didn’t see much growth of the gonad itself so I am still playing around with the exact formula for the culture media. I recently tried out using Shield and Sang’s M3 medium to isolate gonads and in my culture media in addition to the new FBS. Unfortunately, although there was differentiation of gonads in this culture media, the gonads died about 6 hours into imaging. I do believe the medium we had been using, Schneider’s Insect media, is optimal for the differentiation of drosophila embryonic gonads ex vivo. In the future, I plan on doing a titration experiment to find what concentration of FBS is optimal for differentiation and growth in culture media.

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Labwork (PCR)

Following DNA extractions, it was time for amplification of DNA through Polymerase Chain Reaction (PCR). PCR amplifies the DNA by denaturing, copying, and synthesizing over and over again. Denaturing occurs via temperature, first during the DNA extraction process, and thenĀ again using the Thermocline machine (or PCR machine). The thermocline helps to regulate temperature allowing for PCR to go through its different steps, such as synthesis or denaturing. Synthesis occurs using a special polymerase Taq.

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Labwork

Following the collection of data and samples, it was time for lab work to begin. The first step of my procedure was DNA extractions. With around 800 plant samples, I quickly realized financially and time wise doing all 800 would not be practical. This led to many discussions on how to subsample. Was it better to do more transects, with fewer plants from each transect? More plants, with fewer coverage of overall transects? How do you account for the difference in densities between transects? Overtime, with more and more discussions, it became clear that sampling more transects would be a better option, even if that meant fewer plants per transect. Additionally, for any transect with ~30 or fewer plants, the entire transect would be sampled. For any plant over that, a subsample would be done using a random number generator to randomly select which plants should be extracted.

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Fieldwork

The month of June was predominantly spent on fieldwork and preparation for fieldwork. Preparation included obtaining materials, packing and planning ahead for the visits. There are 5 sites for field work: Presquile National Wildlife Reserve (PWR), Blandy Meadows (BLD-M), Blandy Thistle Thicket (BLD-T), Sky Meadows (SKY), and Greenspring (GRN). For PWR, BLD-M, BLD-T, and SKY, overnight visits were required. We were fortunate enough to have access to research housing for all of these. PWR is an island so that required intensive planning as nothing could be forgotten. Once we arrived at each site, we jumped right into fieldwork. [Read more…]