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Updated: Aug 24, 2026
Isolation and Functional Analysis of Mitochondria from Cultured Cells and Mouse Tissue
Published on: March 23, 2015
Isolation and characterization of the mitochondrion-like organelle from Blastocystis hominis
A M A Nasirudeen1, Kevin S W Tan
1Laboratory of Molecular and Cellular Parasitology, Department of Microbiology, Faculty of Medicine, National University of Singapore, 5 Science Drive 2, 117597 Singapore.
Abstract:
Blastocystis hominis in an unusual protozoan parasite of the human intestinal tract. Previous studies have described the presence of mitochondrial-like structures despite the anaerobic nature of the organism. In this study, we describe a simple and rapid technique to isolate and characterize mitochondrion-like organelles (MLO) from B. hominis. The parasite was disrupted using glass beads and the MLO were collected and purified using a sucrose gradient. Negative staining and transmission electron microscopy of the isolated organelles showed mitochondrial-like structures. B. hominis cells were stained with rhodamine 123 and MitoLight to show the presence of transmembrane potential of the MLO. DAPI staining of the cells suggested the presence of DNA in the MLO. Though brief reports have been made in literature, this study is the first to describe a technique for the isolation of the MLO from this organism. Using this technique of isolation, major metabolic functions of the organelle, their associated macromolecules and intra-mitochondrial location can be extensively studied. The role of MLO in this anaerobic protozoan can be widely investigated using this protocol.
Insights
Researchers developed a new method to isolate mitochondrion-like organelles (MLO) from the unusual intestinal parasite Blastocystis hominis. This technique allows for further study of MLO functions in anaerobic protozoa.
Area of Science:
- Microbiology
- Parasitology
- Cell Biology
Background:
- Blastocystis hominis is an anaerobic protozoan parasite found in the human intestinal tract.
- Previous research has indicated the presence of mitochondrial-like structures within B. hominis, despite its anaerobic environment.
Purpose of the Study:
- To develop and describe a simple, rapid technique for isolating and characterizing mitochondrion-like organelles (MLO) from Blastocystis hominis.
- To provide a method for further investigation into the metabolic functions and molecular components of MLO in this organism.
Main Methods:
- Disruption of B. hominis cells using glass beads.
- Isolation and purification of MLO via sucrose gradient centrifugation.
- Characterization using negative staining, transmission electron microscopy, rhodamine 123 and MitoLight staining, and DAPI staining.
Main Results:
- Successfully isolated and purified mitochondrion-like organelles (MLO) from B. hominis.
- Electron microscopy confirmed the presence of mitochondrial-like structures.
- Staining indicated that MLO possess a transmembrane potential and contain DNA.
Conclusions:
- This study presents the first described technique for isolating MLO from B. hominis.
- The developed protocol facilitates extensive study of MLO metabolic functions, associated macromolecules, and intracellular localization.
- This method opens avenues for comprehensive investigation into the role of MLO in anaerobic protozoa like B. hominis.
