Abstract: Doerrier C, Gradl L, Gnaiger E (2019) DatLab 7.4: innovations. Mitochondr Physiol Network 21.16(03)
DatLab 7.4 is the software for high-resolution respirometry (HRR) incorporating the legacy of previous DatLab versions and new, user-friendly features on DL-protocols, DL-Protocol User, 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 7.4 Innovations
Quality control and traceability
Calibration and background source
- For increasing traceability, DatLab 7.4 incorporates the DLD file name of the calibrations and backgrounds applied.
- DL-Protocols can be selected in DatLab 7.4 in the pull-down menu [Protocols]\Run DL-Protocol/Set O2 limit. A DL-Protocol defines the sequence of Events and Marks. By clicking on "Show" a demo DL-Protocol will be opened. Linked to DL-Protocols are templates for storing exported data in a database and for data analysis. A specific DL-Protocol can be assigned to Chamber A or B, or both.
- » Run DL-Protocol/Set O2 limit
Lower O2 limit [μM]
- Lower O2 limits optimized by the Oroboros MitoFit Laboratory are part of the DL-protocols provided. The lower O2 limit 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 [Protocols]\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 B, or both. DatLab 7.4 allows the user to set a default lower O2 limit in a specific SUIT protocol by creating a DL-Protocol User
- » Run DL-Protocol/Set O2 limit
Synchronous DL-Protocol events
- Synchronous DL-Protocol events sets, if ticked, simultaneous events in both O2k-Chambers if the same DL-Protocol is assigned to both chambers.
Instrumental: Browse DL-Protocols and templates
- DatLab 7.4 incorporates a new collection of instrumental DL-Protocols and templates used for maintenance, calibrations and instrumental quality control for O2, AmR, Fluo and MgG applications, without experimental sample in the incubation medium. 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.
- Click on menu [Protocols]\Instrumental: Browse DL-Protocols and templates to open a folder with all the Instrumental protocols provided with the DatLab 7.4 installation. Example traces with instructions and brief explanatory texts and templates for data evaluation can be opened.
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 7.4.
- Click on menu [Protocols]\SUIT: Browse DL-Protocols and templates to open a folder with all the SUIT protocols provided with the DatLab 7.4 installation. DL-Protocols for different mitochondrial preparations and living cellscan be chosen to assess multiple sequences of respiratory coupling control and ET-pathway 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 DL-Protocols, specific mark names are automatically assigned with titration volumes, which are used for calculation of sample dilution in the O2 analysis template DL7.4. DL-Protocol possess unique D### codes and comprise a fixed sequence of events and marks which cannot be changed by the user in a DL-Protocol. However, with DatLab 7.4 the user has more flexibility and the the fixed sequence of events and marks may be edited and/or changed (Skip/Added) in a SUIT protocol and saved as user-specific DL-Protocol DLPU.
Install Oroboros protocol 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).
Browse www.bioblast.at/index.php./Mito-Pedia SUIT
- Link to the web with the library of SUIT protocols and updates is available.
- SUITbrowser is the new tool to find/design the best substrate-uncoupler-inhibitor-titration (SUIT) protocol for your research questions.
Enable DL-Protocol editing and Export DL-Protocol User (*.DLPU)
- DatLab 7.4 offers a new feature in DL-Protocols: flexibility. Fixed sequence of events and marks can be changed (Skip/Added) in a SUIT protocol by the user. Moreover, editions of text, instructions, concentrations and titration volumes of injections in a specific DL-Protocol can be edited and saved as DL-Protocol User (*.DLPU).
- » Export DL-Protocol User (*.DLPU)
Oxygen flux analysis template for DatLab 7.4
- DatLab 7.4 provides a new Excel template which allows data analysis in a time-efficient way.
- » O2 analysis template DL7.4
- 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 7.4 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: H2OH2O, mitochondrial membrane potential, calcium, ATP) and add-on O2k-Modules (ion selective electrodes: TPP+ 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 H2OH2O 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 for 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