Friday, August 29, 2008

Research

So my time in the neurosurgery/ENT lab came to a close last Friday, but given the relatively short time period that I was here (3 months for research, especially when working with biological systems, is so short!), we managed to achieve some good results. During the last 3-4 weeks of my stay, we fixed the cultures of inner ear cells after 13 days of incubation and began to do immunohistochemistical and immunofluorescent staining to determine 1. if the harvested and isolated cells had proliferated and 2. to see if they differentiated into either neuronal cells or hair cells, depending on their origin. We had originally harvested cells from the Organ of Corti, utricle, and spiral ganglion, and then incubated each with a marker for new cell production. This marker is Bromodeoxyuridine (BrdU), which is incorporated into DNA during S phase of the cell cycle, and can thus be used to determine which cells have been newly produced from the beginning of culture. An initial test run with 3-3'-diaminobenzidine (DAB)-developed staining protocols showed many newly produced cells, indicating our culture medium and conditions worked! Many of the cells showed morphology of neurons and hair cells, but this needed to be confirmed with a double-stain for both BrdU and markers specific for neurons and hair cells. We did these using immunofluorescence for BrdU and neuron-specific enolase (NSE) and Myosin-VIIa, for neurons and hair cells, respectively. Amazingly, the stains worked beautifully and showed many cells that stained for both BrdU and the respective cell markers! Given that this was a first-run trial with these cultures, I was actually amazed that it worked. Thankfully, some of the prepwork in testing different antibodies for specificity (we had 3 anti-Myosin-VIIa antibodies from different manufacturers) and experience from the many neurosurgery experiments helped to make this experiment more successful. My PI had also done some of the same types of experiments with mice and guinea pigs, instead of with rats, during this 2-year tenure at Stanford. So although the initial experiments and set-up were successful, more work still needs to be done, and will be continued after my departure. The ultimate goal is to apply these techniques to human tissue, which is harvested post-mortem. And we showed that it is possible to expand and differentiate post-mortem rat tissue, which is a good step in the right direction! And we will be presenting a poster at the University of Bern/Inselspital Day of Clinical Research, so I also was able to get a small publication out the experience, with hopefully more to follow as the work progresses.

Enough with the hardcore science that is specific mostly to my interests! I had a wonderful time working in my lab - it was a wonderful environment, very well-equipped, with staff whom I hope to keep in touch with. As the summer progressed, it became even more international, as at the beginning, it was predominantly me and the Swiss PIs, lab technicians, and neurosurgery and ENT residents (also a German or two thrown in the mix). But the language was almost exclusively German, whereas mid-summer, one of the lab directors, who was out on maternity leave, returned, and as she is French, the language shifted to a mixture of German, English, and French. And while everyone understood every language (except me - no French!), they would almost always speak their own language or English. So Angelique would ask Jasmin a question in French, which she would then respond to in German, and yet everyone could communicate perfectly in that way! Then at the beginning of August, another nationality was thrown into the mix as a visiting PhD student from Denmark joined us, who spoke only English but could understand some German due to the overlap in their languages. The fluidity of language use in Switzerland still continued to amaze me throughout my entire stay.

Being back in the US for a few days now, I miss already hearing the different languages (although I do get a good mixture of Spanish and English in the county hospital here in Houston!), and I miss the more laid-back approach to life. Quality of life is definitely appreciated more in Switzerland, I believe, and I am really missing a good Mittagspause with time to sit down and enjoy lunch . . . it's now back to having 10 minutes or so to wolf down a bite to eat when I have a chance!

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