Talk:MiPNet17.02 PBI-Shredder manual
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Advantages and disadvantages
Advantages
- Standardized tissue preparations for obtaining disrupted cells with functional mitochondria.
- Highly reproducible results.
- Easy handling, especially for beginners.
- Minimum processing time of 10 min.
- Closed Shredder-Tubes ensure safety throughout the entire sample preparation process.
- Either the homogenate may be used directly for HRR, or the homogenization process may be followed by further isolation of mitochondria.
- The homogenate is suitable for optical measurements (e.g. O2k-Fluorometry with safranin for detection of mt-membrane potential) where a homogenous suspension is required.
Disadvantages
- A fraction of mitochondria is lost (c. 50% in our preparations), therefore about twice the amount of tissue is required compared to permeabilized fibres or homogenization of the total tissue with a potter.
- Since not all mitochondria are obtained from the tissue, tissue mass-specific mitochondrial respiratory capacity can be measured only on the basis of additional measurements of a mitochondrial marker (e.g. CS activity) in the total tissue and in the homogenate, to quantify the mt-yield and refer respiration of the homogenate to Ww of tissue.
- Since not all mitochondria are obtained from the tissue, it is difficult to evaluate if specific mitochondrial types are enriched or a representative subsample of all mitochondria is obtained.
- The cytochrome c effect is larger in homogenate compared to permeabilized fibres, hence a small degree of functional impairment of mitochondria is caused by the homogenization process.