PBI-Shredder HRR-Set

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PBI-Shredder HRR-Set

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Description PBI-Shredder HRR-Set: Auxiliary HRR-Tool for tissue homogenate preparation; the Shredder-Kit Box contains the heavy duty high torque SG3 driver with convertible handle, SG3 base with 3 position force setting lever (FSL), battery charger, two lithium ion batteries, Shredder-Tube Cap Tool and Shredder-Tube Ram Tool. The PBI-Shredder HRR-Set includes the Shredder-Kit Box with 100 Shredder-Tubes FT500-PS, 100 Shredder-Tubes FT500-PMS, a pair of sharp forceps for tissue dissection, a pair of scissors and a manual.

OROBOROS INSTRUMENTS: world-wide distributor.

Product ID 13200-04
Type O2k, PBI-Shredder, Catalogue
Link Manual_PBI-Shredder, O2k-Catalogue: PBI-Shredder, Shredder vs. Fibres
Image
PBI-Shredder HRR-Set.JPG

Application video PBI-Shredder

PBI-Shredder HRR-Set consists of

TitleDescriptionProduct idProduct image
PBI-Shredder SG3PBI-Shredder SG3 for tissue homogenate preparation, heavy duty high torque SG3 driver with convertible handle, SG3 base with 3 position force setting lever (FSL), battery charger and two lithium ion batteries. The PBI-Shredder SG3 is included in the PBI-Shredder HRR-Set. Select 230 V or 120 V. OROBOROS INSTRUMENTS: world-wide distributor.52100
PBI-Shredder SG3.jpg
Shredder-Kit BoxShredder-Kit Box: box for storage and shipping, for PBI-Shredder SG352101-01
Shredder-Tube Cap ToolShredder-Tube Cap Tool: component of PBI-Shredder_HRR-Set.52130-01
Shredder-Tube Cap Tool.JPG
Shredder-Tube Ram ToolShredder-Tube Ram Tool: component of PBI-Shredder_HRR-Set.52140-01
Shredder-Tube Ram Tool.JPG
Shredder-Accessory BoxShredder-Accessory Box: 71x335x240 mm inner dimensions, for storage and shipping of Shredder accessories.52201-01
Shredder-TubesShredder-Tubes: consisting of Shredder Tube FT500-PS with Lysis Disk, serrated Shredder-Ram and Shredder-Screw Cap, coral colour (Box of 100) and Shredder Tube FT500-PMS with Lysis Disk, serrated Shredder-Ram and Shredder-Screw Cap, coral colour (Box of 100). 1 box of each is included in the PBI-Shredder HRR-Set.52210-01, 52220-01
Shredder-Tube Analyt Biochem.jpg
Forceps\stainless Steel\straight Tip\sharpForceps\stainless Steel\straight Tip\sharp: for tissue preparation, stainless steel, antimagnetic. One pair is recommended for insertion of the sample into the O2k-Chamber and for handling in combination with Forceps\stainless Steel\rounded Tip\sharp. Set: in HRR-Dissection Set and PBI-Shredder HRR-Set.54210-01
Forcep for tissue preparation straight tip.jpg
Scissors\stainless steel\straight Tip\sharpScissors\stainless steel\straight Tip\sharp, included in PBI-Shredder HRR-Set and HRR-Dissection Set.54310-01
Scissors stainless steel straight Tip sharp.jpg


Instructions

Using the Shredder-Tube Cap Tool, a serrated Shredder-Ram is inserted into one of the Shredder-Tubes, pressing the sample between the serratged 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.


Advantages of the PBI-Shredder

  • Low shear mechanical device for gentle, rapid and safe disruption of tissues;
  • Three position lever for setting reproducible force to the sample during the shredding process;
  • Gentle enough for isolating intact, functional mitochondria;
  • Powerful enough to rapidly break apart difficult samples;
  • Standardized preparations of high quality;
  • Enables reproducible results;
  • Easy handling, especially for beginners;
  • Processing containers:
> Standard tubes for ambient pressure processing
> Closed containers help to ensure safety throughout the entire sample preparation process;
> Excellent for collection, storage, transport and processing
  • High quality of functional mitochondria is obtained from skeletal muscel 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


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 difficulities in reproducing experimental results over time and across laboratories.
  • During manual homogenization it is difficult to know when sufficient tissue disruption has occured - 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


PBI-Shredder-Manual

Preparation of Tissue Homogenates for Diagnosis of Mitochondrial Respiratory Function: MiPNet17.02.
The PBI-Shredder HRR-Set is an auxiliary HRR-Tool providing a standardized approach to prepare homogenates of various tissues with high reproducibility of 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 and warranty information, and the Product Specification Sheet by Pressure BioSciences Inc (PBI).


Application studies with the PBI-Shredder

  • Tissue homogenates for diagnosis of mitochondrial respiratory function: Mouse heart, brain and liver: MiPNet17.15.
  • Mitochondrial respiration in permeabilized fibres versus homogenate from fish liver and heart. An application study with the PBI-Shredder: MiPNet17.03.


List of publications: PBI-Shredder

 YearReferenceOrganismTissue;cellStressDiseases
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.
MiPNet17.03 Shredder vs Fibres2015-06-18
O2k-Protocols
Mitochondrial respiration in permeabilized fibres versus homogenate from fish liver and heart. An application study with the PBI-Shredder.
FishesHeart
Liver
MiPNet18.05 Amplex-Mouse-heart2015-06-16
O2k-Protocols
O2k-Fluorometry: HRR and H2O2 production in mouse cardiac tissue homogenate.
MouseHeart
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
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
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
MiPNet17.02 PBI-Shredder manual2013-11-22
O2k-Manual
PBI-Shredder HRR-Set: preparation of tissue homogenates for diagnosis of mitochondrial respiratory function.
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