On Monday the lung tissue was collected from the bioreactor after running for 45 hours. The lungs were separated into lobes and photographed to document their difference in appearance. (It is important to note here that a mouse has four lobes making up its right lung, but only a single lobe for its left lung.) Half of each lobe was taken for RNA extraction and half of each lobe was prepared for frozen sectioning. While working late on the extraction, I had the opportunity to meet an oncologist who works in the same lab, and we had a nice conversation on clinical oncology, the current state of research, ways in which the MD/PhD programs could be improved, what it takes to start up a lab, etc. His insights were appreciated, and I hope to be able to contact him with any future questions I might have as I continue my PhD research.
In addition to lab work Tuesday I was able to attend medicine grand rounds in which Dr. Muthukumar presented a lecture on "Why Organs Fail? Molecular mechanisms of Fibrosis". His discussion of cellular differentiation complemented what I had been learning in the lab, so it was well worth skipping lunch in order to attend the lecture. Much of the rest of the day was spent reading papers.
On Wednesday I was able to make histology slides from the frozen lung tissue, and I learned that it is best to inject the lungs prior to freezing to ensure they do not fall apart or shred during slicing. Unfortunately, everyone I had spoken to had omitted this step when describing the protocol, so I was left with samples that provided a bit of a challenge. Thankfully, although the process became more time consuming, good sections appear to have been produced, and any slides I make in the future will likely seem easy by comparison. This is probably the best way to learn.
Wednesday evening I was able to attend a second MRI class in which we learned how to scan the head and abdomen. It was very interesting to see the limits of the "breath hold" technique, and I now have a greater appreciation for the work done by programmers in order to reduce motion artifacts. It was nice to learn more about the fundamentals of MRI (T1 vs T2, how to get Susceptibility SPGR images and Axial FLAIR scans, in-phase vs. out-of-phase, etc.). Again, the manual for the course was very easy to follow and made the laboratory portion of the course quite enjoyable.
Thursday I was able to treat the slides with hematoxylin and eosin (H&E) stain then leave them overnight to dry, and Friday I was able to look at the slides and plan out the next stages of the project with my research mentor in better detail.