Friday, July 6, 2012

Week 7-The Research Proposal

I hope everyone had a wonderful Independence Day! 

This week has been action-packed, tension-filled, and stressful! My graduate student mentor, Qiying Fan, is ambitiously preparing for her research proposal and defense that she will soon present to a committee. This is extremely important because it partially defines her Ph.D. candidacy and she must prepare for a variety of questions to be thrown at her--including those from specialized experts hailing from Houston's medical district. Everyone has been trying to help out with her experiments as much as possible in order to allow her adequate preparation time. For practice, she will be presenting her proposal and defense to me. Her presentation includes a lot of information about the initiation and procession of atherosclerosis and the necessity to continue developing the relationship between a particular protein and the onset of atherosclerosis. It also includes a lot of information about the angiotensin II signaling pathway and A Kinase Anchoring proteins. Dr. McConnell told me to critique her analysis and her explanations about her presentation so that she will be better prepared when the real time comes!

One thing that Fan's research presentation showed me was the importance of having a strong connection between the background and the preliminary data. The background indicates the significance of the problem to be examined in the research project while the preliminary data targets  a specific aspect of the problem. In this specific situation, Fan needed to make a stronger connection between her notes about atherosclerosis and the protein that she is investigating in the signal transduction pathway.

Additionally, two graduate students from TSU have been touring the facility lately. Dr. McConnell said that they are prospective lab members and are in the process of learning specific lab protocols/techniques. I was asked to introduce these students to what I've learned over the course of my stay here in the lab, including: autoclaving, filtration, and Western blots. How exciting is that? Just a few weeks ago I was in the process of being trained, and now, I get the opportunity to instruct. It's such a rewarding feeling.

This week I participated in developing Western blots, transferring heart tissue into histology cassettes, and preparing buffers for cardiomyocyte isolation. One buffer I had a fun time making was the Fura 2-AM loading buffer (cardiomyocyte perfusion buffer). It was difficult making this buffer because the substrates do not fully dissolve in the solution unless the pH is adjusted. The contents make the solution basic so hydrochloric acid needs to be added in order to reach proper homogeneity. Fura 2-AM is an intracellular calcium indicator that is ratiometric and UV light excitable. At low concentrations of the indicator, accurate measurements of the intracellular Ca2+ concentration are able to be measured.  
Cardiomyocyte Buffer-Preparation Sheet

Cardiomyocyte Buffer-Preparation

Sonal Performing Cardiomyocyte Isolation



Cori and I
Solution Making

Adjusting the pH

Cori :)

The Finished and Filtered Buffer!

Fan Preparing Her Research Proposal

Fan-Practicing Her Presentation


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