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Abstract: Doerrier C, Gradl L, Passrugger M, Komlòdi T, Mészáros A, Gnaiger E (2018) DatLab 7.3: innovations. Mitochondr Physiol Network 21.16(02)
DatLab 7.3 is the software for high-resolution respirometry (HRR) incorporating the legacy of previous DatLab versions and new, user-friendly features on DL-protocols, quality control, data analysis, documentation, and traceability of measurements with the Oroboros O2k.
• Keywords: DatLab, Innovation DL-Protocols
• O2k-Network Lab: AT_Innsbruck_Oroboros
DatLab 7.3 Innovations
SUIT: Browse DL-Protocols and templates
- A comprehensive set of new SUIT protocols with example traces with instructions and brief explanatory texts and templates for data evaluation can be browsed from inside DatLab 7.3.
- Click on menu [Protocols]\SUIT: Browse DL-Protocols and templates to open a folder with all the SUIT protocols provided with the USB. DatLab protocols (DLP) for different mitochondrial preparations can be chosen to assess multiple sequences of respiratory coupling and substrate states. The DL-Protocol window displays the sequence of pre-programmed events with chemicals, titration volumes, and corresponding final concentrations. These are saved automatically when the event is set. For protocol-specific marks the mark names are automatically assigned, with titration volumes which are used for calculation of sample dilution in analysis templates. Protocol possess unique D## codes and comprise a fixed sequence of events and marks which cannot be changed by the user. However, the user may edit concentrations and titration volumes of injections and store the modified protocol as user-specific DL-Protocol [File]\Export\DL-Protocol User (*.DLPU)]. For user support, we also provide PowerPoint files with explanatory cartoons and excel templates for data evaluation.
- If users wish to alter the nature of the chemicals used or the sequence of injections, we ask them to contact firstname.lastname@example.org.
Install DatLab 7.3 with the Oroboros DL-Protocols package
- The standard Instrumental and SUIT DL-Protocols package is automatically implemented with the simple DatLab programme installation. We recommend a 'clean install': rename your previous DatLab programme subdirectory (e.g. C:\DatLab_OLD).
Instrumental: Browse DL-Protocols and templates
- DL-Protocols for instrumental calibration and maintenance can be browed from inside DatLab 7.3, example traces with instructions and brief explanatory texts and templates for data evaluation can be opened.
- Click on menu [Protocols]\Instrumental: Browse DL-Protocols and templates to open a folder with all the DatLab protocols (DLP) for cleaning, calibration, and background determination provided with the USB. Select a sub-directory and open an example trace and/or template as desired.
- The user is guided stepwise through the protocol (displayed in the DL-Protocol window), facilitating calibration steps, minimizing handling errors and greatly improving performance during O2k measurements.
Lower O2 limit [μM]
- Lower O2 limits optimized by the Oroboros_MitoFit_Laboratory are part of the DL-protocols provided. O2 limits can be individually set for each chamber and modified by the users. When the pre-set value is reached an automatic WARNING is triggered to remind the user that re-oxygenation of the medium may be required.
- To modify the O2 limit, select [Protocos]\Run DL-Protocol\Set O2 limit, tick the box next to Lower O2 limit [µM], enter desired value and click on Apply to chamber A or alternative as needed.
Browse www.bioblast.at/index.php./Mito-Pedia SUIT
- Link to the web with the library of protocols and updates if available
- Numerous statistical functions are available for data analysis. “Marks statistics” organizes the marks and provides flexibility in displaying DL-Protocol marks, user-defined marks or both. The copy-to-clipboard function, with extended settings for backward compatibility, facilitates export to any third-party analysis software.
Compatible with all O2k series
- DatLab 7 is optimized for the new Oroboros O2k-Series H and supports all configurations of the previous O2k-Series.
Calibration and flux analysis
- At the heart of DatLab 7, a conceptual unification is obtained in calibrations and flux analysis of the O2k-FluoRespirometer channels (oxygen and fluorescence: H2O2, mitochondrial membrane potential, calcium, ATP) and add-on O2k-Modules (ion selective electrodes: TTP+ and pH; amperometric modules: nitric oxide or hydrogen sulfide).
O2k channel labels
- Default channel labels can now be changed, and new labels set by the user. E.g., rename the Amperometric channel, Amp, to 'H2O2' for H2O2 measurements by fluorometry; rename the potentiometric channel, pX, to TPP+ for mitochondrial membrane measurements with the O2k-pH ISE-Module.
- For changing the label, go to menu [Oroboros O2k]\O2k channel labels and set the new channel label as desired.
Display and graphs
- Graphs are further improved for real-time display and publication. Versatile options allow an optimal setup of the graphical display of data. Standard graph layouts are provided. Lab and User-defined graph layouts can be stored and organized to support specific applications.
Management of user accounts and experiments
- As a new feature, user accounts are assigned to each measurement for better traceability and customization. This facilitates the organization of experimental data and allows user-specific configurations in DatLab 7. A detailed description section, cohort, sample and subsample data in the “Sample” window improves organization and analysis of results.
Events and marks
- Concentration and volume of the chemicals added during Substrate-uncoupler-inhibition-titration (SUIT) protocols can be stored together with event names and comments. Automatic outlier detection helps to avoid unwanted data points when setting marks during respiratory steady-states. The user has the option to delete, interpolate or restore data points of marks.
- A hyperlink-based contextual help system provides optimal support throughout settings and experimental steps in DatLab 7. MitoPedia provides the background for this function, which warranties up-to-date and verified information anytime.
- The software-driven, feedback-controlled Titration-Injection microPump (TIP2k) is an integral part of the O2k. In DatLab 7 TIP2k control has improved and facilitates user friendly and more flexible programming.
Labels: MiParea: Instruments;methods
HRR: Oxygraph-2k, O2k-Protocol
O2k-Core, DatLab, DL7