Centrifugation Techniques

Cell biology involves the analysis of cells and tissues at both the morphological and biochemical levels in an attempt to understand the molecular mechanisms operating in any living organism. Proper biochemical analysis of biological samples is imperative for constructing complete and true cell biology. The subcellular organization of cells can be studied via electron microscopy or by the separation and subsequent molecular and biochemical analysis of individual cellular components. While electron microscopy provides detailed information about the appearance of cellular organelles, not much information can be obtained regarding their individual functions. As such, a way to separate the different organelles from remaining cellular components was needed (OpenLearn, 2017).

This was made possible by a technique known as cell fractionation which allows for the isolation of various cellular components (nuclear, cytosolic and mitochondrial) based on their subcellular compartmentalized distribution followed by analysis of their molecular information ranging from the state and distribution of a cell or subcellular component to their response to a range of treatments (Dimauro et al, 2012). The separation of cellular components and particles by sedimentation is one of the most powerful tools in biology. Even though sedimentation using centrifugation is not a new technology, it is essential for conducting cutting edge genomic and proteomic research, as it aids in providing purified particles of interest. As such, centrifugation forms one of the most important and widely applied research techniques used in various disciplines today. It services a range of research fields such as biochemistry, cellular and molecular biology while allowing large scale applications across a broad spectrum of industries ranging from large scale commercial applications in the pharmaceutical and agricultural industries to small scale laboratory-scale scientific studies.

Miescher was one of the first researchers to use a centrifuge for the isolation of a cell organelle when he separated nuclei from human pus cells in 1871. A survey conducted at the US National Institutes of Health revealed that over 65% of research workers believe that using centrifugation to purify cells, subcellular organelles, viruses, proteins, and nucleic acids is an essential part of their everyday research, further proving the importance of this technique in today’s scientific world (Sigma-Aldrich, 2011). Current research applications rely on centrifugation for the isolation and harvest of cells, sub...