Introduction
Subcellular fractionation is when the cellular components separate due to ascending centrifugation speeds. Different cellular components can be separated by densities to form a pellet or supernatant after each centrifugation. This process is used to separate nuclei and mitochondria. Nuclei is a very dense organelle in the center of the cell coordinating the cell’s actions: growth, metabolism, protein synthesis, reproduction. The nuclei also will pellet after the 1st low speed of the centrifugation, which can be seen under a microscope. Mitochondria has a low dense organelle that has a double membrane, allowing oxidative phosphorylation. Generates the majority of adenosine triphosphate known as ATP, for energy. The pellet will be formed after the 2nd centrifugation; the absorbance measured of the mitochondria will be low. Succinate dehydrogenase (SDH) is an enzyme located in the mitochondria membrane. FAD is present, it then turns the succinate to cause cellular respiration. SDH will replace with DCIP, which is an artificial electron acceptor. DCIP is used to measure mitochondrial activity in the spectrometer. DCIP is a redox indicator, causing it …show more content…
Table 1 depicts the Velocity of DCIP being reduced by SDH. Table 2 shows a comparison between SDH assay and the images perceived from the microscopy. When looking at table 2, it was seen that Nuclei and mitochondria were only located in the filtrate. S1 contained mostly mitochondria, while P1 contained mostly mitochondria. S2 had some mitochondria while P2 had a majority of mitochondria. When looking at the Azure control column, you can see that the blue color sis not appear in S2 and P2 due to mitochondria being present. The color blue was only present when there were nuclei present. Figure 1 is the filtrate, figure 2 is S1, figure 3 is P1, figure 4 is S2, and figure 5 is P2 under the
Subcellular fractionation is when the cellular components separate due to ascending centrifugation speeds. Different cellular components can be separated by densities to form a pellet or supernatant after each centrifugation. This process is used to separate nuclei and mitochondria. Nuclei is a very dense organelle in the center of the cell coordinating the cell’s actions: growth, metabolism, protein synthesis, reproduction. The nuclei also will pellet after the 1st low speed of the centrifugation, which can be seen under a microscope. Mitochondria has a low dense organelle that has a double membrane, allowing oxidative phosphorylation. Generates the majority of adenosine triphosphate known as ATP, for energy. The pellet will be formed after the 2nd centrifugation; the absorbance measured of the mitochondria will be low. Succinate dehydrogenase (SDH) is an enzyme located in the mitochondria membrane. FAD is present, it then turns the succinate to cause cellular respiration. SDH will replace with DCIP, which is an artificial electron acceptor. DCIP is used to measure mitochondrial activity in the spectrometer. DCIP is a redox indicator, causing it …show more content…
Table 1 depicts the Velocity of DCIP being reduced by SDH. Table 2 shows a comparison between SDH assay and the images perceived from the microscopy. When looking at table 2, it was seen that Nuclei and mitochondria were only located in the filtrate. S1 contained mostly mitochondria, while P1 contained mostly mitochondria. S2 had some mitochondria while P2 had a majority of mitochondria. When looking at the Azure control column, you can see that the blue color sis not appear in S2 and P2 due to mitochondria being present. The color blue was only present when there were nuclei present. Figure 1 is the filtrate, figure 2 is S1, figure 3 is P1, figure 4 is S2, and figure 5 is P2 under the