Difference between revisions of "Biological contamination"

Latest revision as of 16:29, 5 November 2020

high-resolution terminology - matching measurements at high-resolution

Biological contamination

Description

Biological contamination may be caused by microbial growth in the O2k-Chamber or in the experimental medium.

MitoPedia O2k and high-resolution respirometry: O2k-Open Support

Biological contamination of the O2k-Chamber is detected as a high O₂ flux in the closed chamber, measured in a QC1: Oxygen sensor test or QC2: Instrumental O₂ background test.

The problem can be solved by first cleaning the O2k-Chamber by chemical sterilization (70 % ethanol with 30 % water; not 100 % ethanol) and test with a freshly prepared experimental medium, such as MiR05. Make sure the 70% ethanol does not contain any additives; e.g., 70 % ethanol used in hospital settings may contain antiseptics (EtOH containing antiseptics provided by Garcia-Roves PM). If repeated washing and storage with 70 % ethanol does not remove a biological contamination, the glass chamber has to be cleaned as described for protein contamination under O2k-Chamber cleaning with HCl by removing the glass chamber from the O2k and using a strong acid.

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-Biological contamination  is caused by the growing of biological material (originating from samples or otherwise). It should not be confused with other kinds of chamber contamination mentioned in [[Cleaning the glass chamber]].
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+|description=Biological contamination may be caused by microbial growth in the O2k-Chamber or in the experimental medium.
+}}
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+:::: Biological contamination of the O2k-Chamber is detected as a high O<sub>2</sub> flux in the [[Closed chamber |'''closed chamber''']], measured in a [[Oxygen sensor test |'''QC1: Oxygen sensor test''']] or [[Oxygen flux - instrumental background |'''QC2: Instrumental O<sub>2</sub> background test''']].
-Biological contamination of the chamber is typically detected by a high O2 flux at closed chamber near air saturation. It is detected by a [[sensor test]], an [[instrumental background test]] or by observing the flux at closed chamber for a short time routinely before any experiment. In the later two cases the high flux contamination may be caused by biological contamination of the chamber itself or by a contaminated medium. This can be tested by observing the flux in pure water.
+:::: The problem can be solved by first cleaning the O2k-Chamber by chemical sterilization (70 % ethanol with 30 % water; not 100 % ethanol) and test with a freshly prepared experimental medium, such as [[MiR05]]. Make sure the 70% ethanol does not contain any additives; ''e.g.'', 70 % ethanol used in hospital settings may contain antiseptics ([[Talk:MiPNet19.03 O2k-cleaning and ISS#Problem solved |EtOH containing antiseptics]] provided by [[Garcia-Roves PM]]). If repeated washing and storage with 70 % ethanol does not remove a biological contamination, the glass chamber has to be cleaned as described for protein contamination under [[MiPNet19.03 O2k-cleaning and ISS |O2k-Chamber cleaning with HCl]] by removing the glass chamber from the O2k and using a strong acid.
-The ideal counter agent is 70% Ethanol with 30% water (NOT 100% Ethanol). Make sure the 70% ethanol does not contain any additives. E.g.  70% ethanol used in hospital settings may contain antiseptics (information provided by [[Garcia-Roves PM]]).
-Prevention of biological contamination of the chamber is primarily by storage under 70% ethanol. If repeated washing with 70% Ethanol does nor remove an already present biological contamination, the glass chamber has to be cleaned as described for Protein contamination under [[Cleaning the glass chamber]] by removing the glass chamber form the instrument and using a strong acid.
-{{#set: Technical service =Chamber}}
-{{Technical service}}
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