Talk:MiPNet17.02 PBI-Shredder manual

Popular Bioblast page

MiPNet17.02 PBI-Shredder manual has been accessed more than

• 5,000 times (2016-02-25)

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.