Tuesday, June 7, 2011

(II) My Personal Reflection - Jun Hong

1. I choose this project because it is in my area of interest and I would like to learn more if there was a chance.

2. My role in the group is a contributor, blogger, helper and presenter.

3. Challenges we encountered when working on this project the last 2 days was that there was a slacker in our group and we were trying to make him do work. We guided him how to do after much persuasion.

4. Through this project, I discovered new friendships, new people and new knowledge.

5. As an individual, how I have benefited from this programme by listening to what others are saying for fear that I would miss out something important, new knowledge is gained and interacted with people.

6. My aspiration is to have an occupation in the field of Biomedical Sciences, be it Research and Development or a doctor.

My Personal Reflection by Abu Ubaidah

I chose this project basically because I found it extremely interesting. I have a passion for biology and I love biological experiments that could widen my understanding of diseases and how I could actually tell if any organism has such a disease.

My role in the group, I feel, is to provide moral support to my group members and give my all to my project.

Challenges we encountered in the last few days was that we could not meet up often to plan and prepare for the next day. I believe that we overcame this by doing our best whenever we got the chance to meet up.

Through this project, I discovered how my friends from different classes worked together, New ways to discuss and managing time better.

As an individual, this project has vividly increased my interest in the biological field of future occupations and it also benefitted me in many ways by showing me the ways to spot cancer in liver cells.

I aspire to work as a biochemist, researcher or a teacher to inspire future generations to love the study of life.

Abu Ubaidah

(II) Our Mission ~ Part 2

- Histochemical Staining
1) What are the colours of the stained tissue?
Pale pink and Purplish-pink.
2) What is the pattern of colour distribution in the stained tissue?
There are about 25% Purplish-pink and the rest are Pale pink.
3) Can you identify which are the nuclei of the cells? What colour are they?
Purplish-pink round spots are the nuclei of the cells.
4) Can you identify the cytoplasm of the cells? What colour are they?
The pale pink Y-shapes are the cytoplasm of the cells.
5) What are the relative sizes and shapes of the cells in the tissue?

6) How many different types of cells do you think there are in the tissue?
1 type of cell.
7) Are there any other observations you identified? If so, describe them.
The dye was unevenly spread thus could contribute to an unreliable result.

- Immunohistochemical Staining
1) What are the colours in the tissue?
Light brown and a tinge of green.
2) What is the pattern of colour distribution in the stained tissue?
10% brown surrounding the 90% green.
3) Are the colours and pattern of colour distribution similar to that in the first slide?
No.
4) The purpose was to determine the presence and location/distribution of a protein called vimentin. Which colour do you think represents vimentin? Why?
Light brown. Cells produce vimentin thus the surface area of the cell should be surrounded by vimentin and light brown was observed on the surface around the cells.
5) Describe the location/distribution of vimentin in the tissue.
It is distributed unevenly and found in the greatest concentration around the cells.
6) What does the other colour(s) represent?
It represents the cytoplasm of the cells.
7) Are there any other observations you have identified? If so, describe them.
None observed.

- Fluorescence Staining
1) Can you see the tissue using the fluorescence microscope? Does the tissue apear similar to the first and second slide?
The tissue is not visible under the fluorescence microscope and the organelles observed are nothing similar to the first and second slides.
2) Describe what you observe. How many colours appear when using the fluorescence microscope and what are the colours?
Blue and Black colours were observed.
3) What do you think the colours represent? Why?
Blue represents the nuclei of the liver cells while black represents the background and the rest of the organelles of the cells that cannot be stained by the Hoescht dye. The hoescht dye becomes visible as a blue colour under the UV lights at a certain wavelength and since it only attaches to the DNA in the nuclei of the liver cells, the nuclei appears to be blue while the unstained portions of the tissue appears black as UV lights are not visible naturally.
4) Can you determine what is/are the pattern of location/distribution of the colours in the tissue? If no, why not?
No, we do not know how the cells in the liver tissue looks like under the fluorescence microscope as it only shows us the nuclei of the cells.
5) Are there any other observations you have identified? If so, describe them.
Yes, all the nuclei of the liver tissue appears to be of the same shape, This can attribute to identifying different substances quickly.

With reference from Manual for applied challenge project, Dr. Kenneth Yu, Division of Chemical Biology and Biotechnology, School of Biological Sciences, Nanyang Technological University.

Our Gains

This project has indeed deepened our understanding in Microbiology, we learnt in detail on how to use light and fluorescent microscopes. We also learnt on how liver cancer could be diagnosed through immunohistochemical methods by detecting vimentin, which if present in high levels in the liver could attribute to cancer. and how nucleuses could be identified via UV lights emitted through a fluorescent microscope. UV lights can be used at crime scenes as well to determine if blood was indeed real at such a crime scene involving human white blood cells, which has a certain definite shape for its nuclei. Next, the histochemical staining methods gave us a better understanding on how to handle cells in preparation for staining with eosin or hematoxylin, this broadening our knowledge further to understand how to identify cancerous cells at early stages in the liver, a telltale sign being that the liver cells are further apart in the samples. In the future, we might teach others on how to conduct such experiments since we have had our experiences in handling such tissues already, given the facilities necessary.

Monday, June 6, 2011

Plenary Sessions - Reflection by Daniel Pei

Plenary Sessions [Day 1]
(A) My key learning points from each session...
1. Importance of Water - Dirty and Clean: I have learned that water is precious and people are looking into the problems people face with water like lack of proper sanitation and access to clean drinking water.
2. Revolution of Microelectronics Technology: Integrated Circuits can be used to benefit Bio, Radio, Auto, Info, Nano, Water/Hydro, Audio, Video and Energy. 
3. Biomedical Engineering & Technology: Nano technology is about trying to make smaller materials for bigger energy. this can be done by cutting down or building from scratch from the bottom up. Nano technology can be used in many fields like health, defense and virtually. Nano technology can be used in forensic science by detecting chemicals like drugs or explosive on a human finger. 
4. IT for Animation by Prof Seah Hock Soon: There are 2 kinds of animations-2D and 3D. 2D animation costs more for production and is more labour intensive while 3D animation costs less to produce and is less labour intensive.
5. Disappearing glaciers, rising sea levels, and why gravity is even more important than you think: Sea-level rise will not be the same everywhere on Earth. Regions closer to a melting glacier will experience a drop in sea-level and regions furthest away will experience the highest rise in sea-level.
(B) Deepest impression...
The plenary session that leaves the deepest impression on me is Disappearing glaciers, rising sea levels, and why gravity is even more important than you think. What I like about it is that it is an interesting plenary session with touches on a very important subject which concerns the safety and welfare of Singapore and its people. It is about how the melting of the glaciers will affect the sea-levels in various areas, around it and further away from it.  Climate change is a very big threat and we should not ignore it.

Plenary Sessions - Reflection by Abu Ubaidah

The first day of the camp at NTU was indeed insightful. I learnt a lot about a vast variety of fields that are offered or will be offered in NTU in the future. Personally, I found the speakers very engaging and their lectures were very different from the other lectures that I have ever attended, in the sense that they made the talks very interesting indeed and focused on problems at a worldwide level instead of an international one, this making us focus more seriously as they are problems that we could face as a worldwide population. Right now, I shall sum up most of what I have learnt in all the plenary sessions and also the interesting facts I picked up throughout the lectures.
The first lecture about the Importance of Water - Dirty and Clean made me very curious about how the world coped without sufficient water at various places. I realised that if humans actually conserved the water they take every day, They would be enough water for everyone in the world. People are the same no matter what race, No matter what religion and no matter what culture, they still bleed red blood. And as humans, I believe that we should indeed share our water equally with the world. When one makes an effort and he becomes a leader in his fight, He shall inspire generations to follow his beliefs. And with that, I shall say that this speech inspired me in many ways. Not to forget, I shall now tell you about some of my learning points which I would briefly touch on. Firstly, Why water quality is relevant to humans is because it provides protection from diseases and also there is a great need for water for all of us. Next, Water is a large industry of focus in the next few decades as so many problems dealing with water remain unsolved. I also found that many of the things in “Bad” water would lead to death, Environmental degrading, diseases and pungent smells. Finally, I shall use a short sentence to describe all that I have learnt from this talk ; “Water is precious”. 
The next plenary session was about the revolution of microelectronics technology. The talk made me understand that microelectronics and engineering is indeed a vast aspect of life that could be focused on in many different ways. Electronics can encompass virtually and physically everything in life. Be it computers or the human body. Singapore has put in a vast number of money into research for microelectronics, an estimated 16.1 billion dollars. There are also special buildings in Singapore that might just use microelectronics for their independent research. 9 Areas of fields that could be focused on with electronics, if my memory doesn’t fail me are Bio, Radio, Auto, Info, Nano, H2O, Audio, Video and Energy=mc^two. I believe that these aspects solve basically everything in the urban and rural regions so long as subsequent research is put to it. I indeed enjoyed this talk, it made me realise the versatility of jobs that engineering had and also that it helps people tenfold in everything.
The thirds plenary session made me extremely happy as it talked about biotechnology, which is my area of interest currently ( I intend to pursue Biochemistry or Biotechnology). Nanotechnology and Nano-science basically tries to integrate smaller materials to provide bigger energy. It also helps us in many ways, It can detect diseases such as cancer in its primary stages. Actually, even shark teeth had material that could be considered an innovation or nanotechnology of mother nature, the material was small but hard. Biotechnology helps in many ways as well, it brings in fabrication of nano-materials, Forensic sciences and many other interesting things such as artificial photosynthesis. This is indeed a self sustaining industry as research could be put to any need of humans with nano-sciences and biotechnology.
The fourth plenary session was interesting, But I knew most of the details he used beforehand as I have done animation previously in my CCA, Media Club. It was about how long the process of animation could take traditionally. One thing I didn’t know before the talk was that it was harder to make 2D animations rather than 3D. Doing 2D animations a lot, I can imagine the labor involved but I always assumed that 3D was much more complex. 2D animation is much more expensive than 3D animation and even still, Animation makes a big industry to target if you want a job. It also has a vast versatility in jobs that intend to outcome animation, as animation is an extremely tedious and tiring thing. There are script writers , story-boarders, artists, voice actors/actresses, actors/actresses, painters and even computer animators. Indeed an interesting talk by Professor Seah.
The last plenary session about disappearing glaciers made me understand a lot of things, and these realisations come at a global level, being that it can endanger or protect us, depending on what we do now. How melting glaciers causes a decrease in sea levels around the glaciers but an increase in sea levels in the tropics due to complex reasoning such as the formula of Gravitational Attraction (Basically between the glacier and the sea) Fg=Gm1m2/d2. So what does this talk intend to achieve? I think that it wants us to understand that us Singaporeans need to realise sooner or later that we can be in danger with other countries if we do not make an effort to stop Global Warming. When I was younger, I used to be an Environmental Ambassador from the National Environmental Agency because I readily participated in anything that could shape our future. It was indeed insightful, the workshops I have attended and everything, but never have I imagined that there was more to water level rising and dropping than I already knew, I believe this talk also opened up my mind to newer ideas.
I would say that I loved all talks equally but if I were to rank my favourite, I would definitely rate the first plenary session the best along with the last plenary session. Why? I noticed that all the other sessions focused on places in NTU and the brief history of the courses, how NTU could benefit us, but not much of what NTU could do to benefit the world in a problem that currently affects the world. Information from the first and last plenary sessions actually left such a deep impression on me that I started thinking about what me, my friends and Singapore can do to contribute to the wider society, that is, the world. I would like to stress that all the other talks were also informative to me and did provide much detail on what they can solve, but I chose plenary sessions 1 and 5 as their focuses were on a global level.
 That is all,
Abu Ubaidah

(I) Plenary Sessions - Goh Jun Hong

(A) My key learning points from each session...

1. Importance of Water - Dirty and Clean
I learnt that sufficient sanitation facilities and clean drinkable water are needed around the world. We, the Singaporeans, are very lucky whereas in other countries, 1.1 billion people do not have clean water, 2 billion are prone to waterborne disease. Clean sanitation and toilet bowls dates as far back as 1700BC. Asia is predicted to be the largest water environment infrastructure market in the world in the year 2050. Singapore is recognized for dealing with water issues since Singapore has came out with a plan to bring Singapore clean water.

2. Revolution of Microelectronics Technology
I learnt that the start of Microelectronics Technology was started at 1948 due to the invention of the transistors at Bell Laboratories. Electronics play a big role in human life. The invention of integrated circuits and how it changes the electronic industries. The 9 O's: Bio, Radio, Info, Nano, H2O, Audio, Video, E=mctwo. They make the acronym: BRAINWAVE. Microelectronics, according to the professor, is the Most Rounded, Broadest Coverage and Electronics encompasses virtually everything.

3. Innovative Breakthroughs in Nano-Science and Nano-Technology
I learnt that the smaller size the object is that has the same volume, the larger the surface area, thus the more energy. An example the professor provided was that fine sugar dissolved faster than hard sugar because the fine sugar has more surface area thus releasing more energy.
0D - Particles
1D - Fibres
2D - Film

4. IT for Animation
I learnt that stop motion is like story-telling where not much technology is used. In order to animate, Geometric Modeling, Linear Algebra, Calculus and the Laws of Physics have to applied to the movie. 2D animation can look like 3D animation depends of what kind of lighting. According to the professor, Everyone can invent, Identify and Analyze the problem and Invent for a reason. It is the Frontier in Science and Technology, Fascinating Animation. Math is applied in movies and games. IT is in our life.

5. Disappearing glaciers, rising sea levels, and why gravity is even more important than you think
I learnt that the job of a Geodesist is to measure shape and size of Earth gravity field and things that change overtime using space-based techniques. Sea-level rise will not be the same everywhere. Self-attraction and loading is also known as Sea level "Fingerprints". When the glacier is heavy the crust is depressed and water is attracted to the glacier due to gravitational attraction. When the glacier is melted, the crust rebounds and water flows away from the glacier. The formula is "Fg = (G m1 m2)/d^2" which means Gravitational attraction is equivalent to the Mass of one object multiplied to the Mass of another object divided by the square of the distance between the two objects. GRACE or The Gravity Recovery and Climate Experiment is sent to space to observe the Earth's gravitational field and measuring the amount of gravity in each countries.

(B) Deepest impression...
The plenary session that leaves the deepest impression on me is "Disappearing glaciers, rising sea levels, and why gravity is even more important than you think" by Assistant Professor Emma Hill, School of Physical & Mathematical Sciences, Division of Earth Science. What I like is the fact that the presentation was short and sweet, plus easy to understand, while teaching us new terms within the strict time limit of just 15 minutes. Assistant Professor Emma has a loud and clear voice with a tinge of humour, that makes the last plenary session all the more engaging. Geology interest me a lot since it unravels how the Earth actually works, and since it is the first time I heard the term "Geodesist" I was interested in the occupation. I too was interested that gravity actually is an indicator and has a say in Global Warming.

Wednesday, May 18, 2011

Our Accomplishments

Histochemical Staining


Histochemistry is the branch of science concerning tissues and its identification and distribution by means of stains and indicators through chemical and microscopic means. What we did was basically stain colourless liver cells which are hard to view under a normal light with Eosin and Hematoxylin. Through this experiment, we managed to get more insight of the light microscope such as the magnification of the eye pieces, describe different parts of the cells such as the differentiation between the nucleus and the cytoplasm observed in cells after the staining as they appeared in different colours and we also managed to get first hand experience in handling tissues in preparation for histochemical staining. Finally, we also managed to observe different structures of the human liver samples and how they would appear different with cancer (i.e, for liver cells, they appear further apart and less ‘bonded’ as cancer makes the liver grow further with tumors and such.) The colours observed were dark blue for the nuclei, purple and pink for the cytoplasm.



Immunohistochemical Staining Protocol



It is a method to identify a specific protein in cells using a primary antibody that binds to that one particular protein, then a secondary antibody is bonded to the enzyme that targets the first antibody. The enzyme on the second antibody catalyzes a colourless solution to form a colored product, produced by the cells.


A primary antibody, that is targeted to a protein named vimentin, is given to us to be spread on to the liver tissue, and to be incubated for a full hour. It was placed into the buffered solution called PBS. Then a peroxidase-conjugated secondary antibody solution is applied and was left to incubate for 45 mins. It was then again place into PBS. Then an enzyme substrate solution was applied onto it, and was left to incubate for half an hour. Then it was placed in PBS again and rinsed in distilled water. A drop of mounting media is applied and dried for 5 mins. Then we looked into a light microscope to inspect our results.


Some parts of the liver tissue is stained in a light brown colour. If the light brown colour is significant in liver tissue, it means that much vimentin is present, thus it is more likely that cancer cells are present too, since cancer cells excessively produced vimentin. It was then concluded that our liver tissue is most probably infected with cancerous cells.



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Fluorescence Staining


The goal of this technique of staining is to identify and locate organelles in a cell, namely the nucleus and the mitochondria. The dyes can only be viewed though the use of a fluorescence microscope. The microscope splits light into different wavelengths. Fluorescent dyes can only be seen when a specific range of wavelength hits them. Each dye has their specific range of wavelength which excites them to fluoresce and they give out photons which can be seen by us. The dyes used in fluorescence staining sticks to specific organelles like the nucleus or mitochondria. This happens because the dyes used are positively charged and thus, would be attracted to DNA which are negatively charged. DNA is found in the nucleus.


For this project, the objective was to stain the nucleus of liver cells in a slice of liver tissue and observe it under a fluorescence microscope. The dye used to stain the nucleus is the Hoescht dye, which emits the colour blue. The background of images is black. This is due to the UV light that is projected at the DNA of the nucleus/mitochondria of the liver tissue that is binded with the Hoescht dye and it reflects the UV light into our eyes to be seen, while the background is black due to the fact that the background does not have DNA and the Hoescht dye is not binded with it thats why the UV light could not be reflected into our eyes to be seen, thus we see black.



Slides


-Purpose of this method is to identify and locate organelles in a cell through the use of fluorescent dyes and a fluorescent microscope.


-The dyes are positively charged and the DNA, found in the nucleus, are negatively charged.

(II) Our Mission ~ Part 1

This project intends to achieve a deeper understanding on liver cells and how cancer cells may affect them in terms of behavior through the use of staining cells through the use of technology, Namely using immunohistochemistry, histochemistry and fluorescence staining methods.

(I) Overview of Project

This project primarily concentrates on the histology and morphology of cells and for us (the students) to learn and appreciate the usefulness of the light and fluorescence microscope. The project is focused on the histological preparation of tissues. We will then be introduced to the 3 type of histological preparation :

a) Histochemical staining
b) Immunohistochemical staining
c) Fluorescence dye staining

With reference from Nayang Technological University: SST-NTU Flagship Programme: Project 2: Project 2 Manual.pdf

Our Team

A photo of us with Dr Kenneth Yu
Goh Jun Hong - S209

Daniel Pei Zheng De - S208

Abu Ubaidah - S206