In the final week of my internship, we focused on staining the roots we collected the week before and viewing them through the microscope. Monday and Tuesday were spent going through the dyeing process. We began by taking roots out of the test tubes and placing around 5 to 6 in a small orange cassette. Once every cassette was filled we put them into a solution of KOH and put them on a hot plate at 90 degrees celsius for an hour. After the hour was up we moved the roots to a 1% HCL solution for around 20 minutes. The last step for that day was to move the cassettes to a solution of acidic glycerol and trypan blue which we left overnight. When we came back the next day we moved the roots into 1.5 ml microcentrifuge tubes filled with acidic glycerol. We replaced the acidic glycerol 2 more times before the day was up in order to remove the stain from anything that we didn’t want to view under the microscope.
On Wednesday we began to set up the microscope slides. We took the roots from the centrifuge tubes and laid them horizontally on each slide before adding a few more drops of acidic glycerol and putting on the cover slide. Once we completed each slide we went down to a separate lab where we could view the image from the microscope on a computer. Steve taught us how to count the mycorrhizae and what to look for in the microscope. We got to look at a few of the slides before the day was over.
Thursday was spent focusing on microscopy. We viewed slides under the microscope for the majority of the day and began to collect data. In the slides, I saw there was a considerable amount of arbuscular mycorrhizal fungi as well as a pathogen called olpidium. We decided to start counting the olpidium in order to see if the glyphosate had any effect on that.
On my last day in the lab, I put a few finishing touches on the presentation I have been preparing throughout the internship. Once I was ready I presented to everyone in the lab and said my farewells. I had an amazing experience and although I wasn’t able to complete the data collection and analysis I am hoping to view it once it is finished and work remotely with Pacifica. I want to thank everyone who made this possible and everyone I had the pleasure of working with.
During my free time, I visited with a friend who was here from Telluride and explored more of Boulder.
Interesting process and great to see you doing some real science. It was surprising that this research was being done to examine the effects as a negative to using roundup for large scale noxious weed control.