Our mainland travel portion of this experience was started off with a reunion of the entire group in Xiamen. It was really cool to be able to see where the other students had been working and living for the last 7 weeks! We, well the Hong Kong crew as Professor Duncan would put it, even got to see the other students present on their research which was pretty cool (not to mention the after-presentation delicious lunch buffet at a local upscale hotel)!

I really wish we had more time in Xiamen because our five days just flew by! We definitely got to see some really cool sites, such as Gulanyu and South Putuo Temple, but I think the real experience in Xiamen was not the sites but the interactions with its people and its culture. In those 5 days I learned more Mandarian than I had learned in 7 weeks in Hong Kong (well Cantonese). Now this is neither a pro or con but I just mean to say that it was different and I really wish I could have had more time to fully experience it. So before we knew it we were heading to our last destination in China, Beijing.

Beijing was amazing to say the least! I honestly am finding it hard to explain because we got to experience some of the most spectacular views and sites in the entire world in just a few days! We got to explore The Forbidden City, Beihai Park, Tiananmen Square, The Temple of Heaven and the GREAT WALL of China (!!!) just to name a few! The last week and a half of this experience was a marathon ran in 3 seconds! Not to say that I am not thankful for getting to see and experience everything that I did because I am, but I really wish we could have had more time because there was just so much to soak in!

Now I know that I just thanked everyone in my previous post but looking back and reflecting on my experience I cannot thank anyone enough. This truly was an experience of a lifetime and I am very appreciative that I was lucky enough to be chosen to partake in it. So thank you again everyone who made this international experience possible, thank you so very much!

Well my last week in Hong Kong was wonderful!

My weekend started off with a trip to Ocean Park, Hong Kong’s one and only amusement park, with my lab mate, Angel, and some of her friends! I will say that Ocean Park is a definite must-see and I really mean it when I say a must-see because truthfully the rides are in no comparison to American amusement park rides (although my lab mate was still afraid to go on some of them) but the views are just fantastic! Ocean Park is built on a small mountain overlooking the ocean and one of Hong Kong’s most prized beaches, Repulse Bay! And it was pretty cool because one of the coasters even hangs you over the side of the mountain and you really get some nice views! But fantastic views is not the only cool thing that Ocean Park offers, it also has a lot of fun educational exhibits with different types of marine life and when you are a bit of a nerd and a marine science major from Eckerd College you will definitely enjoy yourself! And the icing on the cake is definitely the Panda Bear exhibits! I think Angel could tell how excited I was to see the pandas because she asked me if I wanted my photo taken with them… and of course I did! This trip was amazing and it really was a great start to my last week in HK!

My work week was kicked off by going eel shopping with Oscar at one of the local wet markets! However, in all honesty, I was a bit disappointed because he had told me that we would have to go collect our eels, so as an Eckerd College student I automatically thought that meant that we would have to go into the field (or over the sea wall) and collect the specimens…. But this was not the case. Instead we collected our specimens from one of the local wet markets which was still pretty cool! So once this news was broken to me, I started to wonder how we would choose which eels to buy. Do we just pick the eels that seem the healthiest? Do we actually weigh the eels since we only use eels that weigh between 500-600g? How do we transport them back to HKBU? With all of these questions in my head I decided to risk sounding a bit silly and I asked Oscar, turns out he has to call them in advance because they have to prepare the eels and all we have to do is come and collect them in a huge bag and ride in a taxi back to HKBU…. So this process was a bit easier than I had expected but it was still very cool nonetheless!

As for the rest of my week, it worked like clockwork! We got to run through the entire protocol for two more eels! And I got to have a goodbye lunch with my lab and Professor Wong at the fabulous Dim Sum Restaurant with the famous, wonderful, and delicious milky yellow buns, which they made sure to order for me (very considerate!) J And then it was concluded with Oscar making me try tortoise jelly because it is supposed to be very good for you and apparently it is still even used in Chinese medicine today. Let’s just say I think it is an inquired taste. But it was a perfect ending to my experience in HK because I think it is a perfect example of my overall experience. Living and working in Hong Kong is by no means difficult because there is not much of a language barrier and it’s a very safe and accessible city but then once in awhile you get thrown a curve ball (maybe the menu does not have the English translation or the MTR has already shut down for the night) and it throws you off at first but you learn how to handle it which I think helped prepare us for visiting mainland China (well to an extent).

But before I got into that, I would like to take this opportunity to thank everyone who made this international research experience possible. This experience was absolutely unbelievable.

Thank you very much Professor Duncan, Professor Wong, Oscar and everyone in Professor Wong’s lab (Angel, Becky, Sue, JoJo, Kong, Alice, Milk, Sissi, Bonnie, and Ching), Eckerd College (specifically Professor Flaherty and Professor Cohen) and of course NSF!

But as I said before, I bit off a lot to chew today and I somehow pulled it off! Today I was able to prepare and to perform the protocol for my samples (24 samples!) all the way from their salinity treatments to cDNA! Which is quite a feat if I must say so myself and I think that even Oscar was impressed! Well I am just so thankful that all of my samples came out because these are the freshwater samples that I had to re-do from last week when bacteria decided to crash the party. So in a way I feel that I have redeemed myself from last week’s little slip up J and who knows I may have even exceeded expectations! Well the reason that I say this is because my samples (for both 6 and 24 hour treatments) have thus far (knock on wood!) have come out beautifully! The 260/280 ratios (this can tell you about the quality of your sample), which can be obtained from the nanospectrophotometer, were wonderful! The best range to get is like 1.8 -2.0 and I was getting like 1.98 for most of my samples!!! I was/am so excited! So excited that I decided to have a celebratory ice cream on my way back to my dorm (yes another one! haha)!!

And the best part is that because I was able to convert my samples into cDNA today that on Friday I will be running quite a few real-time PCRs which means a whole lot of results are coming my way! So I will keep you posted!

So this week we dissected again and collected the gills (from both freshwater and saltwater environment eels), performed primary gill cell culture and treated the cells with a smaller range of salinities (because we realized that the extreme salinities were causing too much cell death) for a longer time frame (6 and 24 hours). After the treatments, the idea was that we would perform RNA extraction and convert our RNA samples into cDNA so that way we could re-test the expression levels of NHE-1 and NHE-3 along with other transporters. The only obstacle that stood in the way of this was bacteria!

Yes, unfortunately 2 of the 3 freshwater samples somehow became contaminated and bacteria made its home right smack dab in the middle of all of our beautiful samples. When I saw that all of our samples had turned red in color (because we treat our samples with a pH color sensitive solution, so they normally are a wonderful array colors from yellow to purple) I went from happy and excited to sad and discouraged. But Oscar and Professor Wong were very encouraging and even told me some stories of their own in which bacteria had overtaken their samples too. So I just decided to take Oscar’s usual advice and to not worry about it.

But thankfully the freshwater sample that we prepared for the MTT assay and all three saltwater samples were unaffected! So we were able to continue on with the protocol for the saltwater samples but this week we did not have time to test the transporters. We also got to perform the MTT assay but once again we did not have time to interpret our results (yet). So next week will be another busy week for us in the lab! So overall, I would like to think that this last week as another success. Plus as Professor Wong and Oscar would say that anything that is not considered a “success” is considered experience… so this last week I would like to think that I had both success and experience! And I would like to keep this in mind for the rest of my time here, so that no matter how I get accomplished here in terms of results and in term of progress that I am definitely getting some good experience!!!

The reason why I say this is because all of the hard work, time, and effort that I had put into my research this past week all came down to Friday. My goal was to perform the entire protocol for RNA extraction to cDNA, so this means that I started off this past week from scratch.

So on Monday, I dissected an eel, extracted its gills, and performed the protocol for primary gill cell culture. On Tuesday, I treated the cells with the specific salinities that I wanted to test. I decided to do five different treatments (5pH, 6pH, 7pH, 8pH and 9pH), excluding the control. After treating the cells for 6 hours, I was then able to perform the RNA extraction. Then on Wednesday, we got a break :) The reason is because it was a public holiday! The public holiday is known as the Tuen Ng (Cantonese name) Festival and is most commonly known for its dragon boat races! Gretchen and I were able to actually go watch the dragon boat races at Stanley which was a really neat experience!

But we were back to work on Thursday… So I had to determine the concentration of the RNA for each of my samples which is very nerve wracking! The reason why I say this is because if the concentration isn’t high enough than you can no longer proceed with the procedure and have to start from the beginning with the dissection, so you really only have one shot at this. But luck was on my side and my samples had a high enough concentration! However this is not the end of it yet. On Friday, I then put my samples into the thermocycler to synthesize cDNA from my RNA samples. Then to make sure that everything up to this point has been done correctly and that everything is still good, I had to test the quality of my cDNA by preparing them with a “housekeeping” gene and loading them into the real-time PCR machine. Then after the real-time PCR machine has performed its cycles you can retrieve the cycle number of each of your samples which you can compare with your control to make sure your quality is good enough to continue on with the procedure (plus the cycle number is very important because we will use the cycle number to calculate the level of expression of that specific gene/transporter within the cell which is what we are really interested in).

And this is where the moment of truth comes in….

The moment where you will see if all of the work for the entire week has been done sufficiently enough to actually start the testing (I am pretty sure I had my fingers crossed for the entire PCR cycle which is approximately 2 1/2 hours long! It was especially hard because we were at lunch during this time and eating with chopsticks while your fingers are crossed is not an easy task!) On a side note, the students in my lab think it is actually very funny that we cross our fingers. I did not realize that they do not do it here but I think I may have convinced them to start because… Once again luck was on my side!!! That’s right, my quality of cDNA was good enough to continue on! I cannot even describe how excited I was!! I think Oscar, the graduate student I work with, even commented on how big my smile was!

But this really was fantastic news because I was able to test two different transporters on Friday alone! Plus I am very excited because the amount of cDNA needed to run the test is very a small amount so I still have enough cDNA to run even more tests this week!!! After work I was so happy and everything that I even bought a celebratory ice cream from the local student store for my walk back to the dorms! haha So hopefully (knock on wood), this week will pan out just like last week! Cross your fingers ;)

So on Monday Oscar, the graduate student who I primarily work with, and I dissected an eel and retrieved the gills so that way we could continue on with our research for the week. However, this is not the whole story. This dissection was supposed to be my first, however, Oscar actually had to do the dissection because for some reason the eel did not fully take to the anesthetic and WOKE UP during the dissection! And as you may know an eel’s muscle reflex does not require oxygen, so even if the eel is not alive anymore the muscles can still reflex, which they did and the eel just kept moving! This was quite the experience!

I was a little worried when we had to do some more dissections on Wednesday, but everything went smoothly which was great because this time I had remove the gills! But as for the rest of the week, I learned the protocol for primary gill cell culture which is a very good and useful technique to know! This technique allows us to retrieve the cells we need in order to continue on with our research. From there we then can treat the cells with different salinities and extract the proteins in order to run a western blot. A western blot can then detect specific proteins within our samples. Specifically, this last week we looked for NHE-1 (sodium proton exchanger) and compared it to Actin (comparing to Actin can tell us if I loaded the gel correctly) just as a trial run because it was my first time performing a western blot by myself. I have actually started working very independently in the lab on techniques that I have learned, but I have still a lot of help and support from everyone around me which is just amazing! I should hopefully start working towards my individual research project starting this week so cross your fingers!

And as for the rest of HK, it is as great as usual!

Now being a western student researcher in a Chinese university is quite interesting but I absolutely love it! To start off, my professor is just amazing. He is very understanding, nice, and generous! He has welcomed me with open arms into his lab and I am very appreciative! My first day in the lab he made sure that I was introduced to everyone and that I felt comfortable. Now feeling comfortable in a lab where everyone is welcoming and smiling and where they give you your own personalized lab coat, notebook, and drawer is pretty easy. So far I have worked in the lab less than a week and I have already learned some of the basic techniques that I will be using in the lab and for my project, more specifically I have learned how to do the protocol for: RNA extraction, western blotting, and real-time polymerase chain reaction (PCR). And thus far this week I have started to learn the protocol for the dissection and the primary eel gill epithelium cell culture! Once I have gotten these techniques down I will be meeting with my professor to talk about and figure out my specific topic for my individual project which I am excited to do!

So far I believe that this experience will definitely prepare me for collaborative science with foreign nationals when I am working as a scientist because this research experience is so much more than just the science that we get to learn and partake in in the labs! This research experience thus far is unbelievable.