A tissue homogenate (thom) is obtained through mechanical micro-disruption of fresh tissue and the cell membranes are mechanically permeabilized.
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 »Talk:Tissue homogenate
|BEC 2020.1 doi10.26124bec2020-0001.v1||Gnaiger E et al ― MitoEAGLE Task Group (2020) Mitochondrial physiology. https://doi.org/10.26124/bec:2020-0001.v1||2020|
MitoPedia topics: Sample preparation