MiPNet19.03 O2k-cleaning and ISS: Difference between revisions
From Bioblast
Beno Marija (talk | contribs) No edit summary |
No edit summary |
||
Line 32: | Line 32: | ||
:::: 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]]. | :::: 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 after each experimental use. | |||
| [[Media:O2k-cleaning_BeforeUse.DLP|O2k-cleaning_BeforeUse.DLP]] | |||
|- | |||
| Instrumental O2 background manual injections | |||
| Instrumental O2 background with manual injections including OroboPos test and two-point calibration | |||
| [[Media:Instrumental_O2_background_manual_injections.DLP|Instrumental O2 background manual injections.DLP]] | |||
|- | |||
| Instrumental high O2 background TIP2k | |||
| Instrumental high O2 background with TIP2k including OroboPos test and two-point calibration | |||
| [[Media:Instrumental_high_O2_background_TIP2k.DLP|Instrumental high O2 background TIP2k.DLP]] | |||
|- | |||
| Instrumental high O2 background manual injections | |||
| Instrumental high O2 background with manual injections including OroboPos test and two-point calibration | |||
| [[Media:Instrumental_high_O2_background_manual_injections.DLP|Instrumental high O2 background manual injections.DLP]] | |||
|} | |||
A full list of DL-Protocols (Instrumental DL-Protocols and SUIT DL-Protocols) is displayed in [[DL-Protocols]]. | |||
Revision as of 12:07, 9 November 2017
MiPNet19.03 O2k-cleaning and ISS
O2k-Chamber cleaning SOP and Integrated Suction System (ISS). |
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. ยป Versions
- >> Product: O2k, O2k-Catalogue
โข O2k-Network Lab: AT_Innsbruck_Oroboros
Labels: MiParea: Instruments;methods
HRR: Oxygraph-2k, O2k-Protocol
O2k-SOP
MitoPedia O2k and high-resolution respirometry:
O2k-Open Support
- 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 -20ยฐC. 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.
- ยป Discussion
- 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:
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.
Cleaning DLP
DL-Protocols (DLP) should be used for cleaning the chambers, stirrers and stoppers before and after each experimental use.
DL-Protocol | Description | DLP File |
---|---|---|
O2k-cleaning_BeforeUse | Standard cleaning procedure for O2k chambers, stirrers and stoppers, to be applied after each experimental use. | O2k-cleaning_BeforeUse.DLP |
Instrumental O2 background manual injections | Instrumental O2 background with manual injections including OroboPos test and two-point calibration | Instrumental O2 background manual injections.DLP |
Instrumental high O2 background TIP2k | Instrumental high O2 background with TIP2k including OroboPos test and two-point calibration | Instrumental high O2 background TIP2k.DLP |
Instrumental high O2 background manual injections | Instrumental high O2 background with manual injections including OroboPos test and two-point calibration | Instrumental high O2 background manual injections.DLP |
A full list of DL-Protocols (Instrumental DL-Protocols and SUIT DL-Protocols) is displayed in DL-Protocols.