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| == Additional resources == | | == Additional resources == |
| Β | | Β» [[PBI-Shredder MitoPedia]] |
| Β» [[O2k-Catalogue: PBI-Shredder]] | |
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| Β» High quality of functional mitochondria is obtained from skeletal muscel and kidney as evaluated by high resolution respirometry:
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| :: Gross VS, Greenberg HK, Baranov SV, Carlson GM, Stavrovskaya IG, Lazarev AV, Kristal BS (2011) Isolation of functional mitochondria from rat kidney and skeletal muscle without manual homogenization. Analyt Biochem 418:213-23. - [[Gross_2011_Analyt_Biochem |Bioblast link]]
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| Β» [[MiPNet17.02 PBI-Shredder manual|Manual_PBI-Shredder]] - The PBI-Shredder O2k-Set is an auxiliary O2k-Tool providing a standardized approach toΒ prepare homogenates of various tissues with high reproducibility ofΒ mitochondrial yield and mitochondrial function. In this manual for applications with high-resolution respirometry (HRR), we refer to the PBI User Manual with safety information, product use limitations andΒ warranty information, and the Product Specification Sheet by Pressure BioSciences Inc (PBI).
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| === Application studies with the PBI-Shredder ===
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| Β» [[MiPNet17.15 PBI-Shredder Mouse-heart-brain-liver|MiPNet17.15]] - Tissue homogenates for diagnosis of mitochondrial respiratory function: Mouse heart, brain and liver
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| Β» [[MiPNet17.03_Shredder_vs_Fibers|Shredder vs. Fibers]] - MitochondrialΒ respiration in permeabilized fibers versus homogenate from fish liverΒ and heart. An application study with the PBI-Shredder
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| == Instructions ==
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| :::: Using the [[Shredder-Tube Cap Tool]], a serrated Shredder-Ram is inserted into one of the Shredder-Tubes, pressing the sample between the serrated surface of the Shredder-Ram and the lysis disk of the tube. Ice-cold mitochondrial medium (200-700 Β΅l [[MiR06]] or [[MiR06Cr]]) is then added to the opposite Cap side of the tube, which is closed by a [[Shredder-Screw Cap]], again using the Shredder-Tube Cap Tool.Β The filled Shredder-Tube is placed into the pre-cooled Shredder Base, with the Ram side facing downwards, and the SG3 driver is placed onto the Cap. The force setting lever is placed into the position (1-low, 2-medium), and shredding for 20-30 s is sufficient for several different tissues.Β The processed homogenate is sampled from the Shredder-Tube by unscrewing the Shredder-Screw Cap using the Shredder-Tube Cap Tool and removing the cap, for transfer of the sample into the [[Oroboros O2k|O2k-Chamber]].
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| == Traditional methods for homogenization ==
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| ::::* Homogenizer-based protocols that require extensive operator experience (highly skilled personnel) to obtain reproducible high-quality preparations.
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| ::::* These methods limit dissemination, impede scale-up, and contribute to difficulties in reproducing experimental results over time and across laboratories.
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| ::::* During manual homogenization it is difficult to know when sufficient tissue disruption has occurred - overhomogenization!
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| ::::* The current protocols rely on Dounce, Potter-Elvehjem, and rotor/stator shearing homogenization:
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| :::::: Pallotti F, Lenaz G (2007) Isolation and subfractionation of mitochondria from animal cells and tissue culture lines. Methods Cell Biol 80:3-44. - [http://www.ncbi.nlm.nih.gov/pubmed/17445687 Pubmed]
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| :::::: Frezza C, Cipolat S, Scorrano L (2007) Organelle isolation: functional mitochondria from mouse liver, muscle and cultured fibroblasts. Nat Protoc 2:287-95. - [http://www.ncbi.nlm.nih.gov/pubmed/17406588 Pubmed]
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| :::::: Kaufmann P, TΓΆrΓΆk M, Zahno A, Waldhauser KM, Brecht K, KrΓ€henbΓΌhl S (2006) Toxicity of statins on rat skeletal muscle mitochondria. Cell Mol Life Sci 63:2415-25. - [http://www.ncbi.nlm.nih.gov/pubmed/17013560 Pubmed]
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| == List of publications: PBI-Shredder ==
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| {{#ask:[[Category:Publications]] [[Additional label::PBI-Shredder]]
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| |?Was published in year=Year
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| |?Has title=Reference
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| |?Mammal and model=Organism
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| |?Tissue and cell=Tissue;cell
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| |?Stress
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| |?Diseases
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| |format=broadtable
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| |limit=5000
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| |offset=0
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| |sort=Was published in year
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| |order=descending
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| }}
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| Β | |
| == List of abstracts: PBI-Shredder ==
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| {{#ask:[[Category:Publications]] [[Additional label::PBI-Shredder]]
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| |?Was submitted in year=Year
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| |?Has title=Reference
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| |?Mammal and model=Organism
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| |?Tissue and cell=Tissue;cell
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| |?Stress
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| |?Diseases
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| |format=broadtable
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| |limit=5000
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| |offset=0
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| |sort=Was submitted in year
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| |order=descending
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| }}
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| == Compare ==
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| ::::Β» [[O2k-Publications: Homogenate]]
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