Difference between revisions of "Talk:MiPNet17.02 PBI-Shredder manual"
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== Popular Bioblast page == | == Popular Bioblast page == | ||
[[MiPNet17.02 PBI-Shredder | [[MiPNet17.02 PBI-Shredder manual]] has been accessed more than | ||
:* 5,000 times (2016-02-25) | :* 5,000 times (2016-02-25) | ||
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=== Advantages === | === Advantages === | ||
* Standardized tissue preparations for obtaining disrupted cells with functional mitochondria. | ::::* Standardized tissue preparations for obtaining disrupted cells with functional mitochondria. | ||
* Highly reproducible results. | ::::* Highly reproducible results. | ||
* Easy handling, especially for beginners. | ::::* Easy handling, especially for beginners. | ||
* Minimum processing time of 10 min. | ::::* Minimum processing time of 10 min. | ||
* Closed Shredder-Tubes ensure safety throughout the entire sample preparation process. | ::::* 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. | ::::* 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. | ::::* 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 === | === 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. | ::::* 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, 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. | ::::* 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. | ::::* 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. | ||
Revision as of 07:46, 24 August 2016
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.
