Mitochondrial preparations

high-resolution terminology - matching measurements at high-resolution

Mitochondrial preparations

Description

Mitochondrial preparations (mtprep) are isolated mitochondria (imt), tissue homogenate (thom), mechanically or chemically permeabilized tissue (permeabilized fibers, pfi) or permeabilized cells (pce). In mtprep the plasma membranes are either removed (imt) or mechanically (thom) and chemically permeabilized (pfi), while mitochondrial functional integrity and to a large extent mt-structure are maintained in incubation media optimized to support mitochondrial physiological performance. According to this definition, submitochondrial particles (smtp) are not a mtprep, since mitochondrial structure is altered although specific mitochondrial functions are preserved.

Abbreviation: mtprep

Communicated by Gnaiger E 2011-11-15 (last edit 2022-12-11).

MitoPedia: mt-preparations

» Compare: Submitochondrial particles, smtp

Mitochondrial preparations

General remarks:

Isolation of mitochondria with well preserved function is a prerequisite for the measurement of mitochondrial respiration. In the literature a wide diversity of buffers with different composition for sample homogenization can be found as well as various settings as related to centrifugation force and time for separation of mitochondria. The appropriate method has to be selected to meet specific needs for purity and functionality of isolated mitochondria, additionally taking into account time and cost requirements of the procedure. Functional mitochondria can be prepared only from fresh and not from frozen tissue. It is pivotal that during the entire isolation procedure the samples and buffers are kept on ice.

Specific remarks:

Mitochondria from brain and liver can be separated by simple differential centrifugation of tissue homogenate. Bovine serum albumin should be used to bind free fatty acids released from membranes during the isolation procedure, as these may act as uncouplers. For isolation of mitochondria from heart and skeletal muscle, a protease should be added in the homogenization step so as to release mitochondria from the interfibrillar space.

The listed MiPNet procedures for isolation of mitochondria from rat tissues (brain, liver and heart) were evaluated in the Pharmacobiochemical Laboratory, 3rd Department of Internal Medicine, Medical Faculty, Comenius University in Bratislava, Slovak Republic as optimal for a good yield of mitochondria with satisfactory purity and well preserved function for measurements of mitochondrial respiration.

Protocol	View	Topic	Preparation	Tissue;cell	Organism	Last update
MiPNet21.14 Reference sample HRR	»Versions	Development of a reference sample for high-resolution respirometry.	Intact cells Permeabilized cells	HEK	Human	2024-12-06
MiPNet11.05 imt-pti-pce	»Versions	Isolated mitochondria or permeabilized tissues and cells.	Permeabilized cells Permeabilized tissue Isolated mitochondria			2022-04-10
MiPNet20.06 IsolationMouseHeart-mt	»Versions	Laboratory protocol: isolation of mouse heart mitochondria.	Isolated mitochondria	Heart	Mouse	2021-08-09
MiPNet21.17 BloodCellsIsolation	»Versions	O2k-Protocols: Isolation of blood cells for HRR.	Intact cells Permeabilized cells	Blood cells Platelet	Human	2020-10-22
MiPNet18.05 Amplex-Mouse-heart	»Versions	O2k-Fluorometry: HRR and H₂O₂ production in mouse cardiac tissue homogenate.	Homogenate	Heart	Mouse	2019-06-24
MiPNet20.08 IsolationRatLiver-mt	»Versions	Laboratory Protocol: isolation of rat liver mitochondria.	Isolated mitochondria	Liver	Rat	2016-11-18
MiPNet21.13 Comparison mt-preparations	»Versions	O2k-Protocols: Mitochondrial preparations for HRR.	Permeabilized cells Homogenate Isolated mitochondria	Heart	Mouse	2016-10-18
MiPNet20.07 IsolationRatBrain-mt	»Versions	Laboratory Protocol: isolation of rat brain mitochondria.	Isolated mitochondria	Nervous system	Rat	2016-06-18
MiPNet20.15 IsolationRatHeart-mt	»Versions	Laboratory protocol: isolation of rat heart mitochondria.	Isolated mitochondria	Heart	Rat	2016-03-25
MiPNet14.14 PermeabilizedFiberPreparation	»Versions	Preparation of permeabilized muscle fibers for diagnosis of mitochondrial respiratory function.	Permeabilized tissue	Heart Skeletal muscle	Human	2015-06-22
MiPNet17.15 PBI-Shredder Mouse-heart-brain-liver	»Versions	Tissue homogenates for diagnosis of mitochondrial respiratory function: Mouse heart, brain and liver.	Homogenate			2015-06-22
MiPNet17.02 PBI-Shredder manual	» Versions	PBI-Shredder HRR-Set: preparation of tissue homogenates for diagnosis of mitochondrial respiratory function.	Homogenate			2015-06-19
MiPNet17.03 Shredder vs Fibers	»Versions	Mitochondrial respiration in permeabilized fibers versus homogenate from fish liver and heart. An application study with the PBI-Shredder.	Homogenate	Heart Liver	Fishes	2015-06-18
MiPNet20.09 IsolationBeefHeart-mt	»Versions	Laboratory protocol: isolation of beef heart mitochondria.	Isolated mitochondria	Heart	Bovines	2015-06-15
MiPNet08.13 mt-Isolation-RLM	»Versions	Laboratory protocol: Isolation of rat liver mitochondria.	Isolated mitochondria	Liver	Rat	2010-08-03

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References

Bioblast link	Reference	Year
BEC 2020.1 doi10.26124bec2020-0001.v1	Gnaiger E et al ― MitoEAGLE Task Group (2020) Mitochondrial physiology. Bioenerg Commun 2020.1. https://doi.org/10.26124/bec:2020-0001.v1	2020

Labels:

Preparation: Isolated mitochondria, Permeabilized tissue, Permeabilized cells, Homogenate

MitoPedia topics: Sample preparation