Cookies help us deliver our services. By using our services, you agree to our use of cookies. More information

Difference between revisions of "MiPNet19.03 O2k-cleaning and ISS"

From Bioblast
Line 1: Line 1:
{{Publication
{{Publication
|title=[[Image:O2k-Protocols.jpg|right|80px|link=O2k-Protocols|O2k-Protocols]] O2k-Chamber cleaning SOP and Integrated Suction System (ISS). [[Media:MiPNet19.03 O2k-cleaning and ISS.pdf|»Bioblast pdf«]]
|title=[[Image:O2k-Protocols.jpg|right|80px|link=O2k-Protocols|O2k-Protocols]] O2k-Chamber cleaning SOP and Integrated Suction System (ISS).  
|info=[http://www.bioblast.at/index.php/File:MiPNet19.03_O2k-cleaning_and_ISS.pdf Versions]
|info=[[File:PDF.jpg|50px|link=http://wiki.oroboros.at/images/d/df/MiPNet19.03_O2k-cleaning_and_ISS.pdf |Bioblast pdf]] »[http://www.bioblast.at/index.php/File:MiPNet19.03_O2k-cleaning_and_ISS.pdf Versions]
|authors=OROBOROS
|authors=OROBOROS
|year=2015-11-11
|year=2015-11-11

Revision as of 11:00, 16 February 2016

Publications in the MiPMap
O2k-Protocols
O2k-Chamber cleaning SOP and Integrated Suction System (ISS).

» Bioblast pdf »Versions

OROBOROS (2015-11-11) Mitochondr Physiol Network

Abstract: Fleischmann S, Gnaiger E (2015) O2k-Chamber cleaning SOP and Integrated Suction System (ISS). Mitochondr Physiol Network 19.03(03):1-4.

>> Product: O2k, O2k-Catalogue


O2k-Network Lab: AT_Innsbruck_OROBOROS


Labels: MiParea: Instruments;methods 





HRR: Oxygraph-2k, O2k-Protocol 

O2k-SOP 

Supplement

A properly assembled chamber can remain in the oxygraph for a long time. However to avoid the chamber getting stuck due to dried media and to detect damages or contaminations we recommend to disassemble, clean and reassemble the chamber at least every 1 - 2 years.

General cleaning instructions

There are at least three fundamentally different kind of contaminations that can accumulate in the measuring chamber of the oxygraph and cause problems. All of them have to be treated in different ways:

  • Biological contamination: The ideal counter agent is 70% Ethanol with 30% water (NOT 100% Ethanol). If this does not help the biological contamination may be embedded in e.g. protein contamination and the glass chamber has to be disassembled and cleaned as described below:
  • Protein contamination and other macroscopic deposits. After long use a whitish deposit can form at the glass walls of the chamber. Additionally, small glass splinters (difficult to see) may become embedded in such a deposit. This will be detected by a jumping stirring bar or a stirring bar getting stuck. In this case the glass chamber should be removed from the oxygraph and treated with concentrated hydrochloric acid at least over night. Use a stirring plate and the O2k stirring bar, in this way the stirring bar is cleaned simultaneously.
  • Contamination by hydrophobic inhibitors: The ideal counter agent for this case is pure ethanol.
  • Cleaning with cells, tissue or isolated mitochondria: If there are indications of carry-over of inhibitors or uncouplers, it is recommended to wash the chambers with cells, tissue or isolated mitochondria stored at -20°C. Inhibitors will be bound to the mitochondria when filling the chamber up to the rim such a suspension. If live cells are available, these are preferred due to their superior binding capacity. The suspension should be left in the chambers with inserted stoppers for at least 30 min followed by cleaning the chambers according to the SOPs.


Further information >>


Unintentional introduction of inhibitors to the O2k-Chamber

Inhibitors may also be introduced unintentionally, one example being 70% ethanol used in hospital settings containing antiseptics. Such inhibitors may accumulate e.g. in plastic parts and inhibit subsequent experiments. See the discussion page.

Further details

>> O2k-Chamber
>> MiPNet12.08 O2k-Calibration
>> MiPNet14.06 InstrumentalO2Background