MitoPedia: Sample preparations

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MitoPedia: Sample preparations

MitoPedia - high-resolution terminology - matching measurements at high-resolution.
The MitoPedia terminology is developed continuously in the spirit of Gentle Science.

See also

» O2k-Protocols: O2k-Mitochondrial preparations

Blood cell preparationbcpBlood cell preparation (bcp) is one of the key steps in diagnostic protocols.
Blood plasmaPlasmaBlood plasma is the non-cellular component of the blood. Plasma lacks cellular components of the blood, RBC, WBC and platelets. However, there are many proteins in plasma, i.e. fibrinogen, albumin and globulin. Both blood plasma and PRP maintain clotting activity after whole blood separation.
Blood serumSerumBlood serum is a purified plasma in which the coagulant components were removed from the blood plasma. It contains other substances, i.e. antibodies, antigens and hormones. Serum can be obtained by collecting the liquid phase after blood or plasma coagulation.
Isolated mitochondriaimtIsolated mitochondria, imt, are mitochondria separated from a tissue or cells by breaking the plasma membranes and attachments to the cytoskeleton, followed by centrifugation steps to separate the mitochondria from other components.
Mitochondrial concentrationCmtEMitochondrial concentration is CmtE = mtE·V-1 [mtEU·m-3]. mt-Concentration is an experimental variable, dependent on sample concentration.
Mitochondrial contentmtENXMitochondrial content per object X is mtENX = mtE·NX-1 [mtEU·x-1].
Mitochondrial densityDmtESpecific mitochondrial density is DmtE = mtE·mX-1 [mtEU·kg-1]. If the amount of mitochondria, mtE, is expressed as mitochondrial mass, then DmtE is the mass fraction of mitochondria in the sample. If mtE is expressed as mitochondrial volume, Vmt, and the mass of sample, mX, is replaced by volume of sample, VX, then DmtE is the volume fraction of mitochondria in the sample.
Mitochondrial preparationsmtprepMitochondrial preparations (mtprep) are isolated mitochondria (imt), tissue homogenate (thom), mechanically or chemically permeabilized tissue (permeabilized fibres, pfi) or permeabilized cells (pce). In mtprep the cell membranes are either removed (imt and smtp) or mechanically (thom) and chemically permeabilized (pfi), while the mitochondrial functional integrity and to a large extent the mt-structure is maintained. According to this definition, submitochondrial particles (smtp) are not a mtprep.
Permeabilized cellspcePermeabilized cells (pce) are mitochondrial preparations obtained by selectively permeabilizing the plasma membrane (e.g., with digitonin), for the exchange of soluble molecules between the cytosolic phase and external medium, without damaging the mt-membranes.

Permeabilized cells (pce) are, therefore, not any longer viable or intact cells (ce), since the intactness of cells implies the intactness of the plasma membrane. Any typical quantiative cell viability test (trypan blue etc) evaluating the intactness of the plasma membrane, yields a 100% negative result on fully permeabilized cells.

For permeabilizing the cell plasma membranes chemically with digitonin, without damaging the mt-membranes, the optimum concentration of digitonin must be previously determinated. The protocol SUIT-010 is designed for the evaluation of optimum digitonin concentration for permeabilizing cells, a requirement to account for differences between cell types, the concentration of cells, and variability between batches of the natural product digitonin.
Permeabilized muscle fibrespfiPermeabilized muscle fibres (pfi) are used as a mitochondrial preparation in respirometry to access mitochondrial function comparable to isolated mitochondria (imt). pfi are obtained by selectively permeabilizing the plasma membrane mechanically and chemically (saponin), for the exchange of soluble molecules between the cytosolic phase and external medium, without damaging the mt-membranes.
» MitoPedia topic: Mitochondrial preparations
Permeabilized tissueptiPermeabilized tissue (pti, see also permeabilized muscle fibres, pfi) are mitochondrial preparations obtained by selectively permeabilizing the plasma membrane mechanically or chemically (e.g., with saponin), for the exchange of soluble molecules between the cytosolic phase and external medium, without damaging the mt-membranes.
Platelet-rich plasmaPRPPlatelet-rich plasma (PRP) is obtained as the upper layer at low-speed centrifugation (around 150-200 g), when white and red blood cells sediment and thus get separated from plasma containing the platelets. For further details see blood cell preparation.
SampleA sample is one or more parts taken from a system and intended to provide information on the system, often to serve as a basis for evaluation of the system (diagnosis) or for a decision on intervention (therapy, production process). Compare: Primary sample.
Sample mass concentrationCmXSample mass concentration is CmX = mX·V-1 [kg·m-3].
Submitochondrial particlessmtpSubmitochondrial particles (smtp) consist of membrane fragments which retain most of the enzymatic machinery required in electron transfer and oxidative phosphorylation. Such membrane fragments are continuous closed vesicles formed by resealing of mt-membrane fragments after disruption of the mitochondrial structure. smtp are used to isolate the inner-membrane-bound ET-pathway (mETS) from the upstream modules of the Electron transfer-pathway (ETS) which are located in the mt-matrix and outer mt-membrane (transporters). smtp are obtained by treatment of mitochondria with membrane-dispersing agents such as digitonin at high concentration or by sonic irradiation.
Tissue homogenatethomA tissue homogenate (thom) is obtained through mechanical micro-disruption of fresh tissue and the cell membranes are mechanically permeabilized.

O2k-Publications in the MiPMap

» Isolated mitochondria
» Permeabilized cells
» Permeabilized tissue