Alexandra Rios's blog

I just finished my last week at CMOP! I spent week 10 working on my final paper and powerpoint. I ended the week by giving a final talk and saying goodbye to everyone in my lab. 

This is been yet another amazing internship! I am extremely happy I was able to do it for a second summer. Maybe I'll be back next year - who knows! :)

I cannot believe this internship is coming to an end! I spent this last week wrapping up my project at the bench: I extracted DNA from B.pacifica one last time and amplified genes that will reveal the identity of the bryozoan we are working with. This is being done to be sure that we are, in fact, working with B.pacifica and not a similar bryozoan of the Bugula spp. I sent those sequences off on Monday (7/27/09) and those results should come back to us by the end of the week (just in time to throw it into my presentation/paper!) 

We finally completed bryostatin labeling on my B.pacifica samples! My mentor and I continually put it on our "to  do" list but another experiment always got in the way. Anyway, over last week, I finished bryostatin labeling on B.pacifica larvae and ovicells, and B.neritina larvae. I need to spend more time looking at negative controls before I can come to any conclusions about my positive samples... but let's just say it looks like we have some exciting results (look for them in my presentation / paper)!

I spent this past week taking images of the B.pacifica and B.neritina ovicells that contained (what seems to be) E.sertula cells! It has been quite a challenge to keep everything organized -- I relabeled every sample I considered significant so that I could keep better track of my "final" images.

Also, this week I looked at my final FISH samples from B.pacifica. So far, I have some great images that I am very excited to share in my presentation and paper!

This past week I did FISH on B.pacifica and looked at my samples under the confocal microscope. To my surprise, I found a lot of bacterial cells that appear to be related to E.sertula! It was an exciting week - I took lots of images of what I found and marked each ovicell I looked at for future inspection (I need to be able to estimate each ovicell's developmental stage). My ultimate goal is to show a timeline of ovicell development followed by images that show the bacterium's location within the ovicell.

I am just over halfway done with my internship! I can't believe how quickly it flies by...

I spent the beginning of week 5 running PCRs and purifying the PCR products from B.pacifica. Now I have a few tubes ready to be utilized in cloning, a process that will allow us to sequence the bacterial genes present in B.pacifica to make sure E.sertula exists in this organism.

This past week was particularly exciting because my frontline mentor, another graduate student, and I went to the Oregon Institute of Marine Biology (OIMB) for 2 days. OIMB is another research facility located on the coast near Coos Bay. There, we collected B.pacifica, another bryozoan that is closely related to B.neritina but is typically more difficult to work with. Also, judging from the number of publications online on B.pacifica, this bryozoan has been given less attention than B.neritina.

I realized this week that the B.neritina ovicells I was looking at did not contain very well developed embroys. E.sertula is more likely to be found inside ovicells that contain developed or developing embyros -- empty ovicells probably won't contain any bacteria at all! I spent the majority of the week correcting my mistake by doing FISH on ovicells that clearly contained a well developed embryo. I discarded all empty looking ovicells!

This week, Andrew was able to locate some B.neritina ovicell samples, so now I am well on my way with my project! It is my job to locate ovicells that are "full," do FISH on those samples, and localize E.sertula. By looking at ovicells at different developmental stages, we will be able to better understand E.sertula's movement within the ovicell prior to the release of the larvae. It is important to note that much work has been done, especially by Dr. Haygood and associated researchers, on the location of E.sertula during the larval stages and beyond.

I am happy to say I am back for a second summer internship at CMOP! I am working in the same lab, Dr. Haygood's, and with the same mentor, Andrew Han. Although I am working with the same people and in the same environment, I have an entirely new and unique project to tackle!