PBI-Shredder O2k-Set

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PBI-Shredder O2k-Set


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O2k-Open Support


MitoPedia O2k and high-resolution respirometry: O2k-Open Support 



Description

PBI-Shredder HRR-Set.JPG

PBI-Shredder O2k-Set: Auxiliary O2k-Tool for tissue homogenate preparation


Reference: Product details and purchasing information

Specifications

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): Manual PBI-Shredder


Instructions

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 O2k-chamber.

Video support for PBI-Shredder

  • Instructions on how to use the PBI-Shredder and how to prepare homogenates of various tissues can be found here: Shredder tissue homogenate

Additional resources

» High quality of functional mitochondria is obtained from skeletal muscle and kidney as evaluated by high resolution respirometry:

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. - Bioblast link

Application studies with the PBI-Shredder

» MiPNet17.15 - Tissue homogenates for diagnosis of mitochondrial respiratory function: Mouse heart, brain and liver

» Shredder vs. Fibers - Mitochondrial respiration in permeabilized fibers versus homogenate from fish liver and heart. An application study with the PBI-Shredder


Traditional methods for homogenization

  • Homogenizer-based protocols that require extensive operator experience (highly skilled personnel) to obtain reproducible high-quality preparations.
  • These methods limit dissemination, impede scale-up, and contribute to difficulties in reproducing experimental results over time and across laboratories.
  • During manual homogenization it is difficult to know when sufficient tissue disruption has occurred - overhomogenization!
  • The current protocols rely on Dounce, Potter-Elvehjem, and rotor/stator shearing homogenization:
Pallotti F, Lenaz G (2007) Isolation and subfractionation of mitochondria from animal cells and tissue culture lines. Methods Cell Biol 80:3-44. - Pubmed
Frezza C, Cipolat S, Scorrano L (2007) Organelle isolation: functional mitochondria from mouse liver, muscle and cultured fibroblasts. Nat Protoc 2:287-95. - Pubmed
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. - Pubmed



List of publications: PBI-Shredder

 YearReferenceOrganismTissue;cellStressDiseases
MiPNet18.05 Amplex-Mouse-heart2019-06-24
O2k-Protocols
O2k-Fluorometry: HRR and H2O2 production in mouse cardiac tissue homogenate.
MouseHeart
Bajzikova 2019 Cell Metab2019Bajzikova M, Kovarova J, Coelho AR, Boukalova S, Oh S, Rohlenova K, Svec D, Hubackova S, Endaya B, Judasova K, Bezawork-Geleta A, Kluckova K, Chatre L, Zobalova R, Novakova A, Vanova K, Ezrova Z, Maghzal GJ, Magalhaes Novais S, Olsinova M, Krobova L, An YJ, Davidova E, Nahacka Z, Sobol M, Cunha-Oliveira T, Sandoval-Acuña C, Strnad H, Zhang T, Huynh T, Serafim TL, Hozak P, Sardao VA, Koopman Werner JH, Ricchetti M, Oliveira PJ, Kolar F, Kubista M, Truksa J, Dvorakova-Hortova K, Pacak K, Gurlich R, Stocker R, Zhou Y, Berridge MV, Park S, Dong L, Rohlena J, Neuzil J (2019) Reactivation of dihydroorotate dehydrogenase-driven pyrimidine biosynthesis restores tumor growth of respiration-deficient cancer cells. Cell Metab 29:399-416.MouseEndothelial;epithelial;mesothelial cell
Genital
Other cell lines
Cancer
Haegler 2017 PLOS ONE2017Haegler P, Grünig D, Berger B, Terracciano L, Krähenbühl S, Bouitbir J (2017) Hepatic effects of pharmacological doses of hydroxy-cobalamin[c-lactam] in mice. PLOS ONE 12:e0171026.MouseLiver
Khalifa 2017 Physiol Rep2017Khalifa AR, Abdel-Rahman EA, Mahmoud AM, Ali MH, Noureldin M, Saber SH, Mohsen M, Ali SS (2017) Sex-specific differences in mitochondria biogenesis, morphology, respiratory function, and ROS homeostasis in young mouse heart and brain. Physiol Rep 5. pii: e13125.MouseHeart
Nervous system
Aging;senescence
Cardiovascular
Other
Torres 2016 Redox Biol2016Torres S, Matías N, Baulies A, Nuñez S, Alarcon-Vila C, Martinez L, Nuño N, Fernandez A, Caballeria J, Levade T, Gonzalez-Franquesa A, Garcia-Rovés P, Balboa E, Zanlungo S, Fabrías G, Casas J, Enrich C, Garcia-Ruiz C, Fernández-Checa JC (2016) Mitochondrial GSH replenishment as a potential therapeutic approach for Niemann Pick type C disease. Redox Biol 11:60-72.Human
Mouse
Nervous system
Liver
Endothelial;epithelial;mesothelial cell
Fibroblast
Other
Ejaz 2016 Diabetes2016Ejaz A, Martinez-Guino L, Goldfine AB, Ribas-Aulinas F, De Nigris V, Ribó S, Gonzalez-Franquesa A, Garcia-Roves PM, Li E, Dreyfuss JM, Gall W, Kim JK, Bottiglieri T, Villarroya F, Gerszten RE, Patti ME, Lerin C (2016) Dietary betaine supplementation increases Fgf21 levels to improve glucose homeostasis and reduce hepatic lipid accumulation in mice. Diabetes 65:902-12.MouseFatDiabetes
Kontro 2016 Thesis2016Kontro H (2016) Mitochondrial function and sirtuin expression in hippocampus of young and old high and low-capacity runner rats. Master's Thesis p35.RatNervous systemAging;senescence
Pokkunuri 2016 Oxid Med Cell Longev2016Pokkunuri I, Ali Q, Asghar M (2016) Grape powder improves age-related decline in mitochondrial and kidney functions in Fischer 344 rats. Oxid Med Cell Longev ID 6135319:8p.RatKidneyAging;senescence
MiPNet20.12 IOC103 Manaus2015-07-06
O2k-Network
Manaus BR, 2015 Jul 06-10. To the heart of the Amazon. O2k-Workshop on HRR and O2k-Fluorometry, IOC103.
MiPNet17.02 PBI-Shredder manual2015-06-19
O2k-Manual
PBI-Shredder HRR-Set: preparation of tissue homogenates for diagnosis of mitochondrial respiratory function.
MiPNet17.03 Shredder vs Fibers2015-06-18
O2k-Protocols
Mitochondrial respiration in permeabilized fibers versus homogenate from fish liver and heart. An application study with the PBI-Shredder.
FishesHeart
Liver
Canto 2015 Curr Protoc Mouse Biol2015Cantó C, Garcia-Roves PM (2015) High-resolution respirometry for mitochondrial characterization of ex vivo mouse tissues. Curr Protoc Mouse Biol 5:135-53.MouseHeart
Skeletal muscle
Nervous system
Liver
Fat
Burtscher 2015 Mitochondrion2015Burtscher J, Zangrandi L, Schwarzer C, Gnaiger E (2015) Differences in mitochondrial function in homogenated samples from healthy and epileptic specific brain tissues revealed by high-resolution respirometry. Mitochondrion 25:104-12.MouseNervous systemNeurodegenerative
Hasli 2015 Master Thesis2015Hasli PR (2015) Characterization of mitochondrial respiration and quality differences in diploid and triploid Atlantic salmon (Salmo salar L.) at 5 °C, 10 °C and 15 °C. Master Thesis:1-147.FishesHeart
Skeletal muscle
Temperature
Kluckova 2015 Methods Mol Biol2015Kluckova K, Dong L, Bajzikova M, Rohlena J, Neuzil J (2015) Evaluation of respiration of mitochondria in cancer cells exposed to mitochondria-targeted agents. Methods Mol Biol 1265:181-94.MouseEndothelial;epithelial;mesothelial cell
Genital
Other cell lines
Cancer
Beauchamp 2015 Biosci Rep2015Beauchamp B, Thrush AB, Quizi J, Antoun G, McIntosh N, Al-Dirbashi OY, Patti ME, Harper ME (2015) Undernutrition during pregnancy in mice leads to dysfunctional cardiac muscle respiration in adult offspring. Biosci Rep 35. pii:e00200.MouseHeart
Liver
Cardiovascular
Diabetes
Obesity
Larsen 2014 Anal Biochem2014Larsen S, Kraunsøe R, Gram M, Gnaiger E, Helge JW, Dela F (2014) The best approach: homogenization or manual permeabilization of human skeletal muscle fibers for respirometry? Anal Biochem 446:64-8.Human
Mouse
Skeletal muscle
Gross 2011 Anal Biochem2011Gross 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. Anal Biochem 418:213-23.RatSkeletal muscle
Kidney

List of abstracts: PBI-Shredder

 YearReferenceOrganismTissue;cellStressDiseases
Irving 2013 Abstract MiP20132013Irving BA, Stahl R, Kane CJ, Chernousov M, Argyropoulos G, Carey D (2013) Sex differences in murine mitochondrial oxidative capacity following a 24 week high-fat diet. Mitochondr Physiol Network 18.08.MouseSkeletal muscle
Liver
Fat
Obesity
Krumschnabel 2013 Abstract MiP20132013Krumschnabel G, Eigentler A, Fontana-Ayoub M, Draxl A, Fasching M, Gnaiger E (2013) Tissue homogenates for respiratory OXPHOS analysis in comparative mitochondrial physiology: mouse and trout – heart and liver. Mitochondr Physiol Network 18.08.Mouse
Fishes
Heart
Nervous system
Liver
Eigentler 2012 Abstract Bioblast2012Eigentler A, Draxl A, Fontana-Ayoub M, Gnaiger E (2012) The PBI-Shredder - an auxiliary HRR-tool for the preparation of tissue homogenates for diagnosis of mitochondrial respiratory function. Mitochondr Physiol Network 17.12.MouseHeart
Nervous system
Liver

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» O2k-Publications: Homogenate



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