Discussion Questions from your Mentor

Upload the questions that were posed by your mentor and answer them here.

Histochemical Staining

1. What are the colours of the stained tissue?

     -Pink and blue. 

2. What is the pattern of the colour distribution in the stained tissue?

     -The nucleus is blue, whereas the cytoplasm is pink. 

3. Can you identify which are the nuclei of the cells? What colour are they?

     -Yes, Blue.

4. Can you identify the cytoplasm of the cells? What colour are they?

     -Yes, Pink.

5. What are the relative sizes and shapes of the cells in the tissue?

     -The cells tend to clump together. The cells are irregular in shape and the nucleus is relatively oval in shape. The nucleus is about 1/3 the size of the cells. Adhesion between the cells are bad.

6. How many different types of cells do you think there are in the tissue?

     -I think that there are two, one being the the cancer cells, the other being the regular tissue.

7. Are there any other observations you have identified? If so, describe them.

     -No, as the magnification was too small to see.

Immunohistochemical Staining

1. What are the colours in the tissue?

     -Varying shades of brown.

2. What is the pattern of colour distribution in the stained tissue?

     -Brown precipitate streaks.

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?

     -Brown, as the antibodies recognizes the vimentin and substrate stains it brown. 

5. Describe the location/distribution of vimentin in the tissue.

     -At the border of the cells.

6. What does the other colour(s) represent?

     -The other organelles in the cells.

7. Are there any other observations you have identified? If so, describe them.

     -Some cells were not stained, vimentin is not evenly distributed.

Fluorescence Staining

1. Can you see the tissue using the fluorescence microscope? Does the tissue appear similar to the first and second slide?

     -Yes, the nucleus was glowing blue. No. 

2. Describe what you observe. How many colours appear when using the fluorescence microscope and what is/are the colours?

     -One colour was observed and it was blue. 

3. What do you think the colour(s) represent? Why?

     -The colour blue represented the nucleus as it was stained and the UV light caused it to glow blue. 

4. Can you determine what is/are the patterns of locations/distribution of the colour(s) in the tissue? If no, why not?

     -No, as we were only able to see the nucleus as it was the only part that was stained. 

5. Are there any other observations you have identified? If so, describe them.

     -No. 

Our Achievements

This post documents the learning points and the project finding or the end-product. 


Photographs/Videos that you have recorded during the project. 

Tissue viewed under light microscope after histochemical staining.

Tissue viewed under light microscope after immunohistochemical staining.

Tissue viewed under fluorescence microscope after fluorescence staining.

And a picture of us working on the tissue. 

What have you achieved through the completion of this project? When answering this question, consider both personal and academic development.

Personal:

     -Our attitude was important, as we had to be very patient for the cells to be stained, and it was also very tedious, as some unfortunate accidents might happen, and everything would go wrong. If that happened, then we would have to redo it and it would take up a lot more of our time, where we would have wasted all our time prior. Thus with this, we were able to cooperate well along with our student mentor to complete the tasks. 

Academic:

     -We learnt what cancerous tissues were, how they looked like, how are they formed, and of course their impacts. We also learnt how to operate a light and fluorescence microscope, and appreciate that they even exist. With the help of our student mentor, we were able to locate the cancerous parts of the tissue using 3 different staining methods, and will be able to apply these methods in our future studies. 

How has the design of your experiment or the design of your product allowed you to achieve the project's desired outcomes.

     -The project was well planned, with the equipments all prepared for us. We also had the help of our student mentor throughout, and the instructions were easy to follow, thus we were able to complete the desired outcomes. 

What are some limitations to your project through the consideration of the design of your experiment or the design of your product. 


     -Our tissue was relatively small thus working on it was quite an obstacle, and we were only able to study on 1 type of tissue.



Suggest possible improvements that could be made in order to minimize the limitations that you have identified.


     -Give us a bigger tissue to work on and allow us to study more than 1 type of cancerous tissue. 

Objectives

What does the group need to accomplish by the end of the project?


     - We must be able to prepare the cancerous tissue for viewing under the light and fluorescence microscopes and be able to view the cells. We have to be able to operate both microscopes properly, and identify which parts of the tissue/cell is cancerous.

How will you know whether or not your project has been successful?

     - We would be able to see the nuclei and cytoplasm if we were successful, using histochemical, immunohistochemical, and fluorescence dying. 

Overview

Read the introduction, objectives and theoretical background to the project and then answer the following questions:

What main theory or concept is the project based on?

     -It is based on the study of cells and analyzing them under a microscope and identifying if the cells are cancerous. 

What do you already know about this theory or concept?

• Cells are the basic unit of life
• Consists of a mass which is a living matter called protoplasm.
• Cells consists of nucleus, cytoplasm, cell membrane.
• Plant/Animal cells.
• Prokaryotic/Eukaryotic cells.
• Different functions of cell parts.
• Cancer cannot be cured fully. 
• Cells are 'colorless'.
• We have to 'color' them in order to see the cells.

Our Team

Mohd Adam (S2-03)
Liu Guo Jun (S2-08)
Student Mentor: Esther Cheow :)

Discussion questions from our mentor

Histochemical Staining

1.What are the colours of the stained tissue?

Pink and Blue

2.What is the pattern of the colour distribution in the stained tissue?

3.Can you identify which are the nuclei of the cells? What colour are they?

Blue

4.Can you identify the cytoplasm of the cells? What colour are they?

Pink

5.What are the relative sizes and shapes of the cells in the tissue?

The cells tend to clump together. The cells are irregular in shape and the nucleus is relatively oval in shape. The nucleus is about 1/3 the size of the cells. Adhesion between the cells are bad.

6.How many different types of cells do you think there are in the tissue?

7.Are there any other observations you have identified? If so, describe them.

No, the magnification is too small to see.

Immunohistochemical Staining

1.What are the colours in the tissue?

Varying shades of brown

2.What is the pattern of colour distribution in the stained tissue?

Brown precipitate streaks

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?

5.Describe the location/distribution of vimentin in the tissue.

At the border of the cells

6.What does the other colour(s) represent?

Green is cytoplasm, and brown is vimentin

7.Are there any other observations you have identified? If so, describe them.

Vimentin is not evenly distributed.

Fluorescence Staining

1.Can you see the tissue using the fluorescence microscope? Does the tissue appear similar to the first and second slide?

No

2.Describe what you observe. How many colours appear when using the fluorescence microscope and what is/are the colours?

1, bright blue

3.What do you think the colour(s) represent? Why?

Nuclues

4.Can you determine what is/are the patterns of locations/distribution of the colour(s) in the tissue? If no, why not?

Yes, the fluorescence blue is the nucleus.

5.Are there any other observations you have identified? If so, describe them.

Our Achievements

Immunohistochemical Staining

Histochemical Staining

 Fluorescence Staining