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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.
|Type||O2k, PBI-Shredder, Catalogue|
|Link||PBI-Shredder @OROBOROS, Manual_PBI-Shredder, O2k-Catalogue: PBI-Shredder, Purchase Order @OROBOROS, Shredder vs. Fibres|
PBI-Shredder HRR-Set consists of
|Title||Description||Product id||Product image|
|PBI-Shredder SG3||PBI-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|
|Shredder-Kit Box||Shredder-Kit Box: box for storage and shipping, for PBI-Shredder SG3||52101-01|
|Shredder-Tube Cap Tool||Shredder-Tube Cap Tool: component of PBI-Shredder_HRR-Set.||52130-01|
|Shredder-Tube Ram Tool||Shredder-Tube Ram Tool: component of PBI-Shredder_HRR-Set.||52140-01|
|Shredder-Accessory Box||Shredder-Accessory Box: 71x335x240 mm inner dimensions, for storage and shipping of Shredder accessories.||52201-01|
|Shredder-Tubes||Shredder-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|
|Forceps\stainless Steel\straight Tip\sharp||Forceps\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|
|Scissors\stainless steel\straight Tip\sharp||Scissors\stainless steel\straight Tip\sharp, included in PBI-Shredder HRR-Set and HRR-Dissection Set.||54310-01|
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:
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.
- Frezza C, Cipolat S, Scorrano L (2007) Organelle isolation: functional mitochondria from mouse liver, muscle and cultured fibroblasts. Nat Protoc 2:287-95.
- 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.
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
|Year||Reference||Mammal and model||Tissue and cell||Stress||Diseases|
|MiPNet20.12 IOC103 Manaus||2015-07-06|
|MiPNet17.03 Shredder vs Fibres||2015-06-18||Heart
|MiPNet17.15 PBI-Shredder Mouse-heart-brain-liver||2015-06-16||Mouse||Heart
|Hasli 2015 Master Thesis||2015||Hasli 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.||Temperature|
|Canto 2015 Curr Protoc Mouse Biol||2015||Cantó C, Garcia-Roves PM (2015) High-resolution respirometry for mitochondrial characterization of ex vivo mouse tissues. Curr Protoc Mouse Biol 5:135-53.||Mouse||Heart
|Burtscher 2015 Mitochondrion||2015||Burtscher 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.||Mouse||Nervous system||Neurodegenerative|
|Kluckova 2015 Methods Mol Biol||2015||Kluckova 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.||Mouse||Endothelial;epithelial;mesothelial cell
|Larsen 2014 Anal Biochem||2014||Larsen 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
|Gross 2011 Anal Biochem||2011||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. Anal Biochem 418:213-23.||Rat||Skeletal muscle
List of abstracts: PBI-Shredder
|Year||Reference||Mammal and model||Tissue and cell||Stress||Diseases|
|Krumschnabel 2013 Abstract MiP2013||2013||Krumschnabel 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||Heart
|Irving 2013 Abstract MiP2013||2013||Irving 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.||Mouse||Skeletal muscle
|Eigentler 2012 Abstract Bioblast||2012||Eigentler 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.||Mouse||Heart
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