Update from week 7/8/19

I performed the third trial for the no treatment/ chloroquine experiment. I repeat the same procedures from the previous trials. I perform cell culture, transfection, drug addition, cell fixation, and fluorescence microscopy. I have yet to perform data collection on the trial 3 microscope slides, so I will do this week 7/15/19.

Update for week 7/1/19

The week of 7/1/19, I performed data collection or cell scoring on my Trial 1 of the no treatment/chloroquine experiment. I had to count 100 cells in multiple areas on the slides to see how many of the 100 cells had stress granules and did not have stress granules. I also cultured my cells to make sure that they would still be viable when I entered the lab the following week.

Results from Trial 1/ Starting Trial 2

At the beginning of this week, I looked at my Trial 1 slides for no treatment/ chloroquine that I made the week before. I examined the twelve chloroquine slides and took pictures of those slides. I need to perform statistics on the slides but I will do that throughout this week. In general, the majority of the cells on the chloroquine slides were not viable and the cells that were alive were scarce. One explanation for this could have been forceful washing during the cell fixation process. This week when I do cell fixation, I will make sure to be careful when washing the cells using the autopipette. I have also done cell culture for Trial 2 of no treatment/chloroquine. I will add my DNA conditions to my cells today and add the autophagy inhibitor drug, chloroquine, to the cells tomorrow.

Starting my independent project

This week, I have started my first trial of my independent project. As a reminder, I am trying to determine if pseudophosphatase, MK-STYX is reducing stress granule formation via the autophagy (cell recycling pathway). To tackle this question, I have to culture mammalian cells, transfect the cells with the different DNA plasmid conditions,  induce and inhibit the autophagy pathway via pharmaceutical drugs, fix the cells, and examine the cells under the fluorescence microscope.  This week I am starting my first trial of my experiment looking at the results of added no drug and the autophagy inhibitor drug. Thus far, I have cultured and transfected mammalian cells with my different DNA plasmid conditions. Later in the week, I will add autophagy inhibitor drug, fix the cells, and examine the cells using fluorescence microscopy. I am excited to see preliminary results of this experiment.

Update: Lab Technique Training

Last Friday (June 7th, 2019) was the last day of laboratory training. The first two weeks of the first summer session was to introduce us new trainees to common lab techniques. We learned how to clone DNA via a plasmid preparation process, use to properly perform mammalian tissue culture, set up an experiment in six-well plates, fix cells and mount them on slides, and to examine cells under the fluorescence microscope.  During the first two weeks, we were asked to seriously think about our independent project and how we would execute our first few experiments. So far, I have been enjoying my experience in the lab. Everyday, I learn something new and I can not wait to learn more.