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

From Bioblast
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|journal=Mitochondr Physiol Network
|journal=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.''' Β 
|abstract='''Fleischmann S, Gnaiger E (2015) O2k-Chamber cleaning SOP and Integrated Suction System (ISS). Mitochondr Physiol Network 19.03(03):1-4.''' Β 
:>> Product: [[Oxygraph-2k|O2k]], [[OROBOROS O2k-Catalogue | O2k-Catalogue]]
[[Image:MitoPedia.jpg|left|60px|link=http://www.bioblast.at/index.php/MitoPedia: DatLab|MitoPedia: DatLab]] [[O2k-Chamber MitoPedia]]
{{MiPNet pdf page linking to MitoPedia}}
|mipnetlab=AT_Innsbruck_Oroboros
|mipnetlab=AT_Innsbruck_Oroboros
}}
}}
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{{Technical support integrated}}
{{Technical support integrated}}


:::: 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 barΒ  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 -20Β°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.
::::Β» [[Talk:MiPNet19.03 O2k-cleaning and ISS|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 [[Talk:MiPNet19.03_O2k-cleaning_and_ISS#Apparent_carry-over_of_CI-inhibitors_versus_contamination_in_chemicals_.28update:_solved.29| discussion page]].
== Cleaning DLP ==
[[DL-Protocols]] (DLP) should be used for cleaning the chambers, stirrers and stoppers before and after each experimental use.
{| class="wikitable" border="1"
|-
! DL-Protocol
! Description
! DLP File
|-
| O2k-cleaning_BeforeUse
| Standard cleaning procedure for O2k chambers, stirrers and stoppers, to be applied before each experimental use.
| [[Media:O2k-cleaning_BeforeUse.DLP|O2k-cleaning_BeforeUse.DLP]]
|-
| O2k-cleaning_AfterUse
| Standard cleaning procedure for O2k chambers, stirrers and stoppers, to be applied after each experimental use.
| [[Media:O2k-cleaning_AfterUse.DLP|O2k-cleaning_AfterUse.DLP]]
|-
| O2k-cleaning_AfterUse_stirrers
| Standard cleaning procedure for O2k chambers, stirrers and stoppers, to be applied after each experimental use; including additional stirrer cleaning (for sticky tissues).
| [[Media:O2k-cleaning_AfterUse_stirrers.DLP|O2k-cleaning_AfterUse_stirrers.DLP]]
|-
| O2k-cleaning_AfterUse_stirrers_inhibitors
| Standard cleaning procedure for O2k chambers, stirrers and stoppers, to be applied after each experimental use; including additional stirrer cleaning (for sticky tissues) and cleaning with cell suspension (to remove remnants of inhibitors).
| [[Media:O2k-cleaning_AfterUse_stirrers_inhibitors.DLP|O2k-cleaning_AfterUse_stirrers_inhibitors.DLP]]
|}
A full list of DL-Protocols (Instrumental DL-Protocols and SUIT DL-Protocols) is displayed in [[DL-Protocols]].





Revision as of 14:05, 12 January 2018

                



MiPNet19.03 O2k-cleaning and ISS

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

Β» Bioblast pdf Β» 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.

MitoPedia: DatLab

O2k-Chamber MitoPedia


O2k-technical support and open innovation
Open the pdf document above.
Β» Current O2k-series: NextGen-O2k Series XB and O2k Series J
Β» Current software versions DatLab 8.0: MitoPedia: DatLab


β€’ O2k-Network Lab: AT_Innsbruck_Oroboros


Labels: MiParea: Instruments;methods 





HRR: Oxygraph-2k, O2k-Protocol 

O2k-SOP 


Template NextGen-O2k.jpg


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







Further links

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