MiPNet19.03 O2k-cleaning and ISS

From Bioblast
Jump to: navigation, search
Publications in the MiPMap
O2k-Chamber cleaning SOP and Integrated Suction System (ISS).

» Bioblast pdf

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. » Versions

>> Product: O2k, O2k-Catalogue

O2k-Network Lab: AT_Innsbruck_OROBOROS

Labels: MiParea: Instruments;methods 

HRR: Oxygraph-2k, O2k-Protocol 

Additional: O2k-SOP 

OROBOROS support

O2k-technical support

This information is part of O2k-technical support and open innovation.
A properly assembled O2k-Chamber can remain in the O2k for extended periods of time.
It is recommended to disassemble, clean and reassemble the O2k-Chamber at least every 1 - 2 years, to avoid the chamber from getting stuck, to clean the glass properly, and to detect any possile damages.

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.
» Discussion

Contamination of the O2k-Chamber by inhibitors

Inhibitors may 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 links

» O2k-Chamber
» MiPNet19.18D O2k-calibration
» MiPNet14.06 Instrumental O2 background