Title:

Authors: Johanna Vargas (1201131), Kayleigh Maduro (1190059)

Introduction
The primary aim of this study is to explore phagocytosis, the process where certain white blood cells bind
to and digest large particles. Phagocytosis, seen as a basic defence mechanism, offers insights into immune
function. Analyzing cell count and leukocyte ratio provides an overall assessment of circulating cell numbers,
offering a glimpse into the health of the blood donor.

Research questions
1. What is the number of white blood cells per volume blood?
2. What are the types of cells?
3. What is the ratio between cell types?
4. Which cells are involved in phagocytosis?
5. Is the optimum incubation time 5 or 10 minutes?

Hypothesis
• The pagocythosis process is expected to take an average of 9 minutes (But its highly content dependent).
• The blood cell types to be involved in the process of phagocytosis are neutrophils, macrophages,
monocytes and dendritic cells.
• A healthy cow is expected to have approximately on average 7 X10^6 leukocytes per ml blood

Method(s): generally described (no details)

- Only principal of method
Isolation of leukocytes from whole blood (Bovine blood)
- Pipette 1ml blood in 1 ml Histopaque
- Centrifuge for 10 minutes at 3000 x g
- Pipette the leukocytes of the tube and put them in a new tube (2).
- Wash the cells (adding 1 ml PBS) and then centrifuge for 10 seconds (at maximum speed).
- Pour the supernatant and and resuspend the cells in the remaining liquid by pipettung/flicking +
add 200 microliter of PBS.
Counting cells and calculating the amount of cells in the blood
- Take 10 microliters from the cell suspension of tube 2 int a Burker counting chamber.
- Use a micoscope with the 40X grid.
Phagocytosis of yeast cells
- Making smears from blood.
- Incubation of blood with yeast cells.
Studying phagocytosis by cells in blood in time using light microscopy
Calculate the amount and the types of isolated leucocytes per ml blood.

Results: describe your own results. Thus not what you should have seen, but what you actually did see.
Try to be as quantitative as possible. Feel free to use graphs and/or tables to help you present your results
in a clear way. Visualization by using your own photo’s is also appreciated.

• Number of cells in 25 squares= Amount of cells X 10^4 / ml in the suspension

• 19*10^4/200= 950*5= 4750 number of cells

The principle behind using a density gradient like Histopaque is that different cell types have different
densities, and when a sample is layered on top of the gradient and centrifuged, the cells separate based on
their densities. The specific density of 1.083 g/mL is often suitable for isolating mononuclear cells because
it allows these cells to form a distinct layer in the gradient.
Discussion + conclusion: How reliable are your observations? Are your observations in accordance with
literature? What can be improved? What do you conclude?
Report

At the end of this practical each duo must hand in a report in English containing your findings: at max one
A4. The report must contain the following:

1. Title: free
2. Authors: names and registration numbers.
3. Introduction + hypothesis and/ or research questions
4. Method(s): generally described (no details)
5. Results: describe your own results. Thus not what you should have seen, but what you actually did see.
Try to be as quantitative as possible. Feel free to use graphs and/or tables to help you present your results
in a clear way. Visualization by using your own photo’s is also appreciated.
6. Discussion + conclusion: How reliable are your observations? Are your observations in accordance with
literature? What can be improved? What do you conclude?

Use a scientific way of writing. That means, write objective, concise and precise. Write the report in the
passive voice (e.g. ‘The cells were isolated’ instead of ‘We isolated the cells’).

One student of the duo must upload the report in Brightspace at the folder assessment/ assignment/
practical report phagocytosis. Name of the file must contain both last names of the student duo. Attention:
Do not forget to include both author names + registration numbers on the report!!!