de Souza Wanderley, Morgado-Diaz José Andrés, Cunha-e-Silva Narcisa L
Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
Methods Mol Biol. 2008;425:313-31. doi: 10.1007/978-1-60327-210-0_25.
Cell fractionation, a methodological strategy for obtaining purified organelle preparations, has been applied successfully to parasitic protozoa by a number of researchers. These studies have provided new information of the cell biology of these parasites and have supported investigators to assume that some of the protozoa form the roots of the evolutionary tree of eukaryotic cells. The cell fractionation usually starts with disruption of the plasma membrane, using conditions that minimize damage to the membranes bounding intracellular organelles. An important requirement for successful cell fractionation is the evaluation of the isolation procedure that can be made by morphological and biochemical methods. The morphological approaches use light and electron microscopy of thin section of different fractions obtained, and the biochemical methods are based on the quantification of marker enzymes or other molecules (for instance, a special type of lipid, an antigen, etc.). Here we will present our experience in the isolation and characterization of some structures found in trypanosomatids and trichomonads.
细胞分级分离是一种获取纯化细胞器制剂的方法策略，许多研究人员已成功将其应用于寄生原生动物。这些研究为这些寄生虫的细胞生物学提供了新信息，并支持研究人员假设某些原生动物构成了真核细胞进化树的根基。细胞分级分离通常始于破坏质膜，所使用的条件要尽量减少对界定细胞内细胞器的膜的损伤。成功进行细胞分级分离的一个重要要求是对分离程序进行评估，这可通过形态学和生化方法来实现。形态学方法利用对所获得的不同级分薄切片的光学显微镜和电子显微镜观察，而生化方法则基于对标记酶或其他分子（例如，一种特殊类型的脂质、一种抗原等）的定量分析。在此，我们将介绍我们在锥虫和毛滴虫中发现的一些结构的分离和表征方面的经验。
