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Immunodetection of Outer Membrane Proteins by Flow Cytometry of Isolated Mitochondria
Published on: September 18, 2014
Atomic force microscopy of isolated mitochondria.
Bradley E Layton1, M Brent Boyd
1Applied Computing and Electronics, The University of Montana College of Technology, Missoula, MT, USA. layton@coe.drexel.edu
Methods in Molecular Biology (Clifton, N.J.)
|June 11, 2011
Summary
This study details methods for imaging mitochondria using atomic force microscopy, revealing outer membrane pores in organelles challenged by glucose or dinitrobenzene (DNB). The focus is on the imaging techniques for these vital cellular components.
Area of Science:
- Cell Biology
- Biophysics
- Toxicology
Background:
- Mitochondria are crucial for cellular energy production and are affected by metabolic and toxic challenges.
- Understanding mitochondrial structure, particularly membrane pores, is key to comprehending cellular dysfunction.
Purpose of the Study:
- To describe detailed methods for isolating and imaging metabolically and toxicologically challenged mitochondria.
- To showcase the capabilities of atomic force microscopy in visualizing intact organelle structures.
Main Methods:
- Isolation of mitochondria from rat dorsal root ganglia or brain.
- Exposure of isolated mitochondria to glucose or dinitrobenzene (DNB) to simulate specific cellular conditions.
- Imaging of mitochondria using atomic force microscopy (AFM) to visualize membrane structures.
Main Results:
- Atomic force microscopy provided high-resolution images of intact mitochondria.
- Specific images revealed outer membrane structures, including voltage-dependent, anion-selective channel pores.
- The study includes images of isolated nuclei and notes on common pitfalls in organelle isolation and imaging.
Conclusions:
- Detailed imaging methods for challenged mitochondria using AFM are presented.
- The techniques allow visualization of fine membrane structures on intact organelles.
- This work provides a methodological foundation for further studies on mitochondrial function and dysfunction.
