Vietnam National University, Vietnam
Pham Phuong Ngoc
Hi, Pham Phuong Ngoc here, an actual "greenhorn" in science who has just finished her 4th year at the University of Science, Vietnam National University. Frankly talking, coming to Aoki lab is undoubtedly one of the most precious times in my life until now because I did not know a laboratory could be that fun and professional. To be more specific, Aoki lab has awe-inspiring potential in analyzing and quantifying the raw data of cell signaling from fluorescence imaging. Therefore, in 3 months of working here, I have attained a comprehensive knowledge of using advanced equipment to observe the changes in colors, depending on the intracellular signaling, inside the cells.
My research topic radically focused on creating a stable HeLa cell line with 4 biosensors, detecting the fluctuations of cAMP existence, calcium concentration, RhoA translocation, and the dynamic of the ERK pathway through the activation of G protein-coupled receptor (GPCR).
Drowning in a sea of studies on GPCR, many cell cycle progressions, such as proliferation, differentiation, and cell survival, are shown to be related to a myriad of specific GPCR ligands. However, the mechanism of how signaling dynamics can quantitatively decide the cell's fate has yet to be unraveled. It is also noteworthy that cancer - a leading cause of death worldwide is basically the accumulation of cell cycle abnormalities. For this reason, we believe that visualizing the level of cell signaling after treating cells with different stimulants lays a concrete foundation for cancer research. Correspondingly, this experimental result may provide a good source of kinetic parameters for computer-assisted simulation, a potential approach for predicting the efficiency of medicine or drugs.
In 3 months, I dealt with DNA works to integrate the 3 types of fluorescence into 3 different and appropriate vectors. Then, the fluorescent expression vectors will be transferred into the HeLa cell line. After selecting cells with a vector of choice, fluorescent microscopy was subsequently utilized to examine the changes in fluorophore's color, depending on the cell signaling activities. Particularly, results obtained by time-lapse videos clearly showed me the colorful changes in a living cell. For me, that was incredibly awesome!
Figure 1: Activity of three different biosensor in the same HeLa cell line.
Note: (A) Activity of cAMP biosensor – GreenFalcan 0.3; (B) Activity of RhoA biosensor – dTomato; (C) Activity of ERK biosensor – KTR-iRFP.
During the time pursuing our main goals, I unintentionally made mistakes in experiments. Still, every time, Aoki professor and Aoki lab members were always there to give me a hand and, together with me fixing my problems. Through that, I have learned about so many things which are needed for working in a professional laboratory: carefully scheduling, fastly cracking mistakes, efficiently discussing with other members, and confidently giving and performing personal views of points.
Besides fulfilling my starvation for research with tons of invaluable lessons, I can also further my knowledge about Japanese traditional culture - my most interesting culture from the bottom of my heart since I already have a deep love for the 2D industry.
Touring the streets enveloped in sparkling lights, sampling many traditional delicacies such as Kirimochi or Kakumochi, Yakisoba, Karaage, Ramen, Red and White Miso, Oinari, Monaka, and many, many dishes that I had never tried before. In like manner, visiting a list of stunning temples, shrines, and the Kaiyukan aquarium in Osaka, entering Maid and Owl Coffee, trying the Game center in Osu Market, dipping in the yellow of autumn leaves in Kuragari and Korankei valley, enjoying the Halloween vibe under the Nagoya TV Tower, taking pictures with most creative cosplayers,... all distributed to my memorable moments here. And the most special thing that I cannot forget was my first time watching the Total Lunar Eclipse on November 7 and watching the Leonid Meteor shower at Otogawa river at 3A.M, then together with some of my precious Japanese friends making our wishes. That special day ended by flaking out at my friend's cozy house. We, 3 individuals, somehow slept in just a single bed and woke up at noon.
All in all, I would like to express my sincere thankfulness to my supervisor, Professor Aoki, and all laboratory members, who allowed me to practice and research at the Division of Quantitative Biology, National Institute for Basic Biology (NIBB). I really appreciate their support, guidance, understanding, belief, and, most importantly, encouragement. Consequently, I was trained on how to work/think critically and how to do research on my own.
Last but not least, with all regard and infinite gratitude, my thank would also send to NIBB for granting me a chance to work and study in such a wonderful workforce in my best living condition.
And my last words to all "greenhorns" like me from any parts of the world, I would highly recommend NIBB as your must-try stopover in your academic path and vigorous youth.