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| {{MitoPedia
| | #REDIRECT [[PBI-Shredder O2k-Set]] |
| |description='''PBI-Shredder''': Auxiliary O2k-Tool for [[tissue homogenate]] preparation with high mitochondrial yield and function.
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| |info=[[PBI-Shredder O2k-Set]]
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| }}
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| __TOC__
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| {{Template:Technical support integrated}}
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| » [[O2k-Catalogue: PBI-Shredder]]
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| == Specifications ==
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| ::::The PBI-Shredder O2k-Set includes all the PBI-Shredder components for a standardized approach to prepare homogenates of various tissues with high reproducibility of mitochondrial yield and mitochondrial function. For more information for applications with high-resolution respirometry (HRR)including PBI-Shredder specifications and information (''i.e.,'' safety information, product use limitations and warranty information, and the Product Specification Sheet by Pressure BioSciences Inc (PBI): [[MiPNet17.02 PBI-Shredder manual|Manual_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 µl [[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. 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. 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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| === Video support for PBI-Shredder ===
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| :::* Instructions on how to use the PBI-Shredder and how to prepare homogenates of various tissues can be found here: [https://www.youtube.com/watch?v=EjQGUr5M8v0&feature=youtu.be Shredder tissue homogenate]
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| ==Additional resources==
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| » 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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| === Application studies with the PBI-Shredder ===
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| » [[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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| » [[MiPNet17.03_Shredder_vs_Fibers|Shredder vs. Fibers]] - Mitochondrial respiration in permeabilized fibers versus homogenate from fish liver and heart. An application study with the PBI-Shredder
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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, Török M, Zahno A, Waldhauser KM, Brecht K, Krähenbühl 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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| == 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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| ::::» [[O2k-Publications: Homogenate]]
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