Isolation and Study of Organelles from Pea Seeds
Isolation and Study of Organelles from Pea Seeds by Differential Centrifugation and Corn to Determine Mitochondrial Activity
BIOL 230-01 Cell & Molecular Biology
October 13, 2014
ABSTRACT
Since the discovery of the cell in 1665 by Robert Hooke, man has always wondered about the complexity and potential functionality. Using pea seeds, which is readily available, we will be studying the different parts of the cell by separating it into its component parts by cell fractionation while still maintaining the properties of the individual organelles. Differences in physical characteristics of organelles, such as size, shape, and density, make it possible to separate them by spinning in a centrifuge at various speeds and for different periods of time. This process is appropriately called differential centrifugation. Using raw and boiled corn kernels we compared whether or not they contained mitochondria. In a test tube containing with mitochondria and active dehydrogenases we found a color change supporting the fact of active mitochondria. Raw kernels have active dehydrogenation because of the colorless tetrazolium turning red. We can understand that boiled kernels were absent mitochondrial activity as well as active dehydrogenases due to the absence of any pink areas coincided with our understanding. Overall, the purpose of this lab was to find mitochondrial activity and/or active dehydrogenase using differential centrifugation and cell fractionation.
INTRODUCTION Given the complexity of the cell and the limited techniques available for its exploration it has been useful for scientists to first separate the cell into parts and study each part and its function as a separate entity. This understanding can be applied to the inner workings of the cell as a whole and visualized by microscopic techniques.
With this in mind, in this experiment a homogenate, a mixture of cellular materials, was made from pea
BIOL 230-01 Cell & Molecular Biology
October 13, 2014
ABSTRACT
Since the discovery of the cell in 1665 by Robert Hooke, man has always wondered about the complexity and potential functionality. Using pea seeds, which is readily available, we will be studying the different parts of the cell by separating it into its component parts by cell fractionation while still maintaining the properties of the individual organelles. Differences in physical characteristics of organelles, such as size, shape, and density, make it possible to separate them by spinning in a centrifuge at various speeds and for different periods of time. This process is appropriately called differential centrifugation. Using raw and boiled corn kernels we compared whether or not they contained mitochondria. In a test tube containing with mitochondria and active dehydrogenases we found a color change supporting the fact of active mitochondria. Raw kernels have active dehydrogenation because of the colorless tetrazolium turning red. We can understand that boiled kernels were absent mitochondrial activity as well as active dehydrogenases due to the absence of any pink areas coincided with our understanding. Overall, the purpose of this lab was to find mitochondrial activity and/or active dehydrogenase using differential centrifugation and cell fractionation.
INTRODUCTION Given the complexity of the cell and the limited techniques available for its exploration it has been useful for scientists to first separate the cell into parts and study each part and its function as a separate entity. This understanding can be applied to the inner workings of the cell as a whole and visualized by microscopic techniques.
With this in mind, in this experiment a homogenate, a mixture of cellular materials, was made from pea
References: CITED Gaynor, Dr. Jack, Dr. Reginald Halaby, Dr. Sandra Adams, Dr. James Campanella, and Dr. Quinn Vega. Laboratory Manual: Cell and Molecular Biology BIOL 230 Fall 2011. New Jersey: Department of Biology and Molecular Biology Montclair State University, 2011. Claude, A. (1946). Fractionation of mammalian liver cells by differential centrifugation: I. Problems, methods, and preparation of extract. The Journal of experimental medicine, 84(1), 51. De Duve, C., & Beaufay, H. (1981). A short history of tissue fractionation. The Journal of cell biology, 91(3), 293s-299s.