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The aims of the experiments are to learn how to classify white blood cells into two groups - granulocytes and agranulocytes. Next, to observe the effect of Deoxyribonuclease (DNase) and Ribonuclease (RNase) on nucleic acid. Thirdly, to learn how to use methyl-green pyronin to locate and DNA and RNA in white blood cells. Lastly, practice safe science when handling body fluids such as blood.
One of the other aims of the experiment was on how to prepare a blood smear. Knowing how to prepare a blood smear would be beneficial as scientist could study more about red and white blood cells closely. One example of industrial applications would be staining the blood, as it could check whether patients are suffering from leukemia, as higher amount of white blood cells would be present in patients suffering from leukemia. Another example of application of identification of nucleic acid would be the mass production of insulin for patients who could not produce insulin, as scientist would have to identify a small code of nucleic acids which is called the gene of insulin. Therefore, after identifying, they would abstract it out and attached it to Escheria.coli for replication of insulin genes.
White blood cells, also known as leucocytes, are made in the bone marrow. They help the body by protecting it against viruses, bacteria and diseases. They could also be called the "white armies of the body". There are two major groups of white blood cells which are granulocytes and agranulocytes.
One difference between granulocytes and agranulocytes are their nucleus. Granulocytes have "grainy" nucleus while agranulocytes have large "whole nucleus". Granulocytes consist of neutrophil, basophil, and eosinophil. Each of them has their own functions in helping the immune system. Agranulocytes consists of lymphocyte and monocyte. They too, have their own functions in helping the body.
In every nucleus of a eucaryote cell, large and condensed molecules of nucleotide will be present. They are known as nucleic acids, they also carry genetic information of the living thing. However, there are two types of nucleic acids, one known as deoxyribonucleic acid (DNA) while the next is ribonucleic acid (RNA).
The structure of a DNA is double helix while RNA is single-stranded. DNA could be found in the nucleus only, however, RNA could be found in both nucleus and cytoplasm. Both are different in their functions, DNA helps in cell division while RNA helps in protein synthesis.
Methyl green pyronin is a stain that would help visualize white blood cells easier, allowing scientist to differentiate between granulocytes and agranulocytes. Methyl green has a preferential affinity for DNA, which would stain DNA green, and pyronin for RNA, which stains it rose-red. The higher the intensity of the colour also represents higher concentration level of DNA and RNA.
RNase is an enzyme that would degrade RNA. A cell added with RNase would only contain DNA as all RNA has already been degraded. One functions of RNase is that it is used to clean up cellular RNA.
The hypothesis of the experiment was that the RNase treated slide will only have green coloured nucleus after adding methyl green pyronin (MGP). The slide that is not treated with RNase will have green and red colour.
The results obtained from the experiment was that, slide treated with RNase showed only green nucleus in the white blood cells after stained by methyl-green pyronin. The slide which only was stained by methyl-green pyronin showed two colours in the nucleus which are red and green with an addition of red cytoplasm.
The results also showed the appearance of the white blood cells. Both granulocytes and agranulocytes were observed in the blood smear slides.
The results obtained were the same as the hypothesis. However, the amount of white blood cells present in the blood smear was lower compared to other group's results. One of the reasons might be because due to the time taken for rinsing of slides. No proper stop watch was used and time could be only estimated. Therefore, the time taken was not accurate.
Furthermore, artifacts were observed during the microscopic examination. This could mislead into other type of results, as some type of artifacts might look like a cell. Therefore, in order to have no artifacts at all, the glass slides have to be clean so that no dust particle would appear on the blood. Air bubbles have to be kept out when putting cover slip on glass slides.
One step in methods and materials was supposed to be done, which was adding xylene as it could make the cells clearer to be seen under the microscope. However, this step was not done. The reason is because xylene is carcinogenic and might be harmful towards the human body. Therefore, to avoid any risks, xylene was not used in the experiment.
Some steps could be carried out during the experiment so that the results could be improved. As hair dryer was used during the experiment, this was not encouraged because the heat might damage the cells. Therefore, results would be inaccurate.
One other step could be that the blood smear slides were put in an increasing alcohol concentration. This would allow maximize amount of water to leave the cell, making the cell to its lowest water potential level.
Lastly, bright-field microscopy is best suited to viewing stained or naturally pigmented specimens such as stained prepared slides of tissue sections or living photosynthetic organisms. It is useless for living specimens of bacteria, and inferior for non-photosynthetic protists or metazoans, or unstained cell suspensions or tissue sections (Caprette, 2005).
In conclusion, the aims were achieved as skills such as preparing blood smears and observing DNA and RNA were done. The results were proven as it has the same readings of the hypothesis. Therefore, the experiment was a success.