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Communicated by [[Gnaiger Erich]] last update 2020-10-24
== Sample type, sample and subsample ==
:::: A sample of a defined '''[[sample type]]''' subjected to a specific sample preparation (see [[MitoPedia: Sample preparations]]) may be, ''e.g.'', a mitochondrial preparation obtained from an organ, cells suspended from a cell culture dish, a tissue biopsy, an organ, an individual organism, or a group of organisms (''e.g.'', a number of nematodes studied collectively in an experimental chamber).
::: '''''Example'': Sample and subsample'''
:::: A blood '''sample''' or a biopsy taken from an individual organism. The '''[[sample size]]''', ''N'', corresponds to the number of [[replica]], ''e.g.'', ''N''<sub>org</sub> organisms in a study group (arm). A sample may be processed and split into a number of '''[[subsample]]s''' (''e.g.'', smaller volumes of blood taken from a larger volume of blood) for (i) application of different types of [[assay]], and (ii) a number of [[repetitions]] or technical repeats, ''n'', of the same assay, using subsamples of the same sample.
== ISO 15189:2012 ==
:::: A '''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) ([[ISO 15189:2012 Medical laboratories — Particular requirements for quality and competence]]).
:::: The term 'system' is used in this context with reference to a 'study system', distinguished from the 'system' in the context of thermodynamics (open, closed, isolated systems; instrumental chamber).
:::: Compare: [[Primary sample]].
== Application in [[HRR]] ==
::: The amount of tissue used for high-resolution respirometry strongly depends on the sample preparation: For permeabilized fibers, usually 0.7 mg tissue; for many cell types, 1 Mill cells per mL, however for some cell types it can be less. With the [https://www.oroboros.at/index.php/product/o2k-sv-module/ O2k-sV-Module] (included in all [https://www.oroboros.at/index.php/o2k-products/packages/ O2k-packages]), the amount of tissue required can be decreased by a factor of 4.
'''Addition of sample to the respirometer chamber'''
:::'''[[Isolated_mitochondria|Isolated mitochondria]] (imt):''' Imt are typically added with a Hamilton syringe. Care should be taken to not exert damaging low pressure during aspiration of the sample into the syringe or exerting high pressure during injection into the chamber.
:::'''[[Living cells]] (ce)''' are typically added with a pipette. Care should be taken to not exert damaging low pressure during aspiration of the sample into the pipette or exerting high pressure during injection into the chamber.
:::'''Very large and/or fragile [[living cells]] (ce):''' Some cell types may be easily damaged when injected with a syringe. For these we recommend: remove the stopper, replace a defined volume of the respiration medium with cell suspension, close the chamber again. Sometimes (i.e. [[microalgal]] cells) the whole volume of the chamber (2 mL) is replaced by the cells suspension with the cell concentration adjusted to the desired value. ''Further details'': »[[Cell count and normalization in HRR]].
:::'''[[PBMC]]s and [[Platelet]]s''': We recommend adding for PBMC 2·10<sup>6</sup> x/mL; for PLT 200·10<sup>6</sup> x/mL.
:::'''[[Permeabilized tissue]] (pti)''': For permeabilized tissue, remove the Stopper, add the permeabilized tissue into the O2k-chamber, close the chamber again.
:::'''[[Permeabilized muscle fibers]] (pfi)''': Remove the Stopper, add the permeabilized fibers into the O2k-chamber, close the chamber again.
:::'''[[Permeabilized cells]] (pce)''': If the cells can be loaded into a syringe, they may be injected with a Hamilton syringe. If the cells are rather fragile, remove the Stopper, replace a defined volume of the respiration medium with cell suspension, close the chamber again.
== References ==
| mainlabel=Bioblast link
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