I started this week off with a wonderful 4th of July. I was able to enjoy a ferry ride to Coronado Island with my host family as well as the fabulous parade at Coronado filled with flair and fun. We then had some gyros, took the ferry back to down town San Diego and biked back to the house. Our afternoon was then busy with preparations for our 4th of July celebratory barbecue. After a delicious meal we set out to watch the fireworks. It was amazing, because I was able to see five firework shows!
After the 4th I started my shortened week at the lab. I started off on Tuesday was some data analysis from the data I collected last week using samples of FITC. The data was not we expected, so in order to try and correct this I found the R (reference) value of the fluorometer. (Usually when working with the fluorometer you set the parameters of the machine to graph S/R or signal ÷ reference, this usually makes the data more uniform because it takes out any variability from the lightbulb within the fluorometer.) When we found the R value and multiplied it to get S by itself the data still was not what was expected. In addition, I used two different entrance and exit slit widths in the fluorometer 2/2 nm and 3/3 nm. These two different slit widths gave completely different results, which was unexpected. As a result we decided to run the experiment again and compare the data. I made a wider variety of samples of FITC ranging from 0.2 µM to 10 µM (micro Molar). I found that when just comparing the signal (S) of the new data that they maxed out the fluorometer at about the same cps (counts per second), even though it took the 2/2 nm slits much longer than the 3/3 nm slits to get to the max point.
In addition I ran an experiment using NATA to compare the true concentration vs. the intended concentration using the UV Vis. In this experiment, I found that the UV vis is only accurate up to a certain point and is very precise and accurate for the lower concentrations, with an absorbance of less than 0.5 at 280 λ.
Finally, I finished off the week starting my new experiment using a protein cyt c (Cytochrome C) and a denaturant, Urea. The protein is a beautiful red color, the color of blood, because it has the same heme group as blood. The function of cyt c is to deliver electrons within cells. For this experiment I made 2 stock solutions along with my previously made KPI (Potassium Phosphate) buffer. I made an 8M Urea stock in buffer as well as a 600 µM stock of the cyt C protein. After that, I made blanks for each of the different concentrations of Urea, these blanks had everything but the protein, then I made the actual samples using a mixture of the Urea stock, buffer and the cyt c stock. I made samples for 15 different concentrations of Urea and all their blanks for a total of 30 different samples. At the end of the day Friday I used a refractometer to measure the index of refraction for all 30 samples. This will help me find the concentration of all my samples come Monday.
Outside of the lab:
This week I moved into my new apartment. The commute to work is great, it takes only 15 minutes door to door! I share this space with two other pinterns, as well as some locals. This weekend Karis, Canyon and I (the other pinterns) went to the birch aquarium and then went to the beach. Finally, today I went to the mall, since the apartment is walking distance, and I was able to do some window shopping. I can’t wait to start next week!
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