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The separation of cellular components such as organelles, ribosomes, nuclei and mitochondria from a tissue homogenate is commonly achieved by differential centrifugation. This technique is based mainly on differences in the size, mass, density and sedimentation rate of cellular components. When a tissue is homogenized gently in an appropriate isotonic buffer, the plasma membranes are disrupted while many intracellular structures remain relatively intact. The resulting homogenate contains nuclei, mitochondria, ribosomes, membrane fragments and soluble cytoplasmic components. These components can then be separated by centrifuging the homogenate successively at increasing speeds. The general sequence of separation is from the largest and heaviest structures to the smallest and lightest structures. The sequence is therefore: nuclei → mitochondria → smaller membrane-bound organelles and microsomal fractions → ribosomes → soluble cytoplasmic components. Since the question specifically asks about organelles, ribosomes, nucleus and mitochondria, the important order is nucleus first, mitochondria next, other smaller organelles subsequently, and ribosomes last. Initially, the tissue is homogenized using a homogenizer in a suitable isotonic medium. The isotonic medium prevents excessive swelling or shrinking of organelles and helps __________ ___ ____ _________ __________ ___ ____ ___ _________ ________ __________.
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