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Difference between revisions of "Tissue homogenate"

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{{MitoPedia
{{MitoPedia
|abbr=
|abbr=thom
|description=A '''tissue homogenate''' is obtained through mechanical micro-disruption of fresh tissue and the cell membranes are mechanically permeabilized. Β 
|description=A '''tissue homogenate''' (thom) is obtained through mechanical micro-disruption of fresh tissue and the cell membranes are mechanically permeabilized.
|info=[[MiPNet17.02 PBI-Shredder-Manual]]
|info=[[MiPNet17.02 PBI-Shredder manual]]
|type=Method
}}
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{{MitoPedia topics|type=Method
== Pros and cons ==
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::::* Homogenized tissue (thom) provides various advantages compared to isolated mitochondria (imt) or permeabilized fibres (pfi): the preparation is faster, no detergents (saponin) are required, tissue heterogeneity may entail a statistical problem in application of pfi which is averaged in the thom preparation, oxygen limitation is reduced, smaller amounts of tissue are needed compared to imt. thom is well suited for the study of mt-respiration ([[Pecinova 2011 Mitochondrion]]).
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::::* Mitochondrial markers have to be measured, ''e.g.'' [[citrate synthase]] activity, to obtain respiration independent of mitochondrial density (although [[flux control ratio]]s can be applied equally).
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::::* Disadvantages of thom compared to imt include: (i) The presence of non-mt membranes increases nonspecific binding of indicators of mt-membrane potential. (ii) Cytosolic components of liver tissue interfere with the measurement of H<sub>2</sub>O<sub>2</sub> production using [[Amplex UltraRed]]. (iii) Cytosolic components may scavenge [[ROX]].
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'''Discussion''' Β»[[Talk:Tissue homogenate]]
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== References ==
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|?Has title=Reference
|?Was published in year=Year
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{{MitoPedia topics
|mitopedia topic=Sample preparation
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Homogenized tissue provides different advantages: the preparation is faster, no detergents (saponin) are required, tissue heterogeneity may entail a statistical problem in application of fibres which is averaged in the homogenate preparation, oxygen limitation is reduced, smaller amounts of tissue are needed compared to isolated mitochondria. Tissue homogenates may be well suited for the study of mitochondrial respiration ([[Pecinova 2011 Mitochondrion]]).

Latest revision as of 14:19, 31 August 2020


high-resolution terminology - matching measurements at high-resolution


Tissue homogenate

Description

A tissue homogenate (thom) is obtained through mechanical micro-disruption of fresh tissue and the cell membranes are mechanically permeabilized.

Abbreviation: thom

Reference: MiPNet17.02 PBI-Shredder manual

Pros and cons

  • Homogenized tissue (thom) provides various advantages compared to isolated mitochondria (imt) or permeabilized fibres (pfi): the preparation is faster, no detergents (saponin) are required, tissue heterogeneity may entail a statistical problem in application of pfi which is averaged in the thom preparation, oxygen limitation is reduced, smaller amounts of tissue are needed compared to imt. thom is well suited for the study of mt-respiration (Pecinova 2011 Mitochondrion).
  • Mitochondrial markers have to be measured, e.g. citrate synthase activity, to obtain respiration independent of mitochondrial density (although flux control ratios can be applied equally).
  • Disadvantages of thom compared to imt include: (i) The presence of non-mt membranes increases nonspecific binding of indicators of mt-membrane potential. (ii) Cytosolic components of liver tissue interfere with the measurement of H2O2 production using Amplex UltraRed. (iii) Cytosolic components may scavenge ROX.
Discussion Β»Talk:Tissue homogenate

References

Bioblast linkReferenceYear
Gnaiger E et al ― MitoEAGLE Task Group (2020) Mitochondrial physiology. Bioenerg Commun 2020.1. https://doi.org/10.26124/bec:2020-0001.v12020


MitoPedia topics: Sample preparation