NextGen-O2k Scientific developments

The revolutionary all-in-one instrument to conquer mitochondrial disease.


WP2: Scientific developments

Abstract

The aim of this workpackage is to test measurement conditions for the newly developed modules and the development of specific applications.

• Determination of optimal Q-sensor measurement conditions
• Determination of optimal measurement conditions for the PB-Module
• Development of protocols for scientific, pre-clinical and algal biotechnology applications
• Data handling and quality control

Achievements

Project Year 1: During testing in the first project year, optimal measurement conditions were determined for using the Q-sensor with a variety of biological samples. Simultaneously testing began of the beta version of DatLab. New SUIT protocols were developed for general, pre-clinical, and algal biotechnology applications. A focus has also been given to internal and external quality control measures.

Progress and next steps

2021-03-01: Changes in the NextGen-O2k firmware made for better functioning of the PB-Module at low light intensities.

2021-02-18: Decision made about which prototype of the white PB-Sensors will be tested by the algae KOLs.

2021-02-18: MitoFit preprint publication: Coupling and pathway control of coenzyme Q redox state and respiration in isolated mitochondria

2021-02-11: New NADH-Sensors for measuring NADH autofluorescence in the NextGen-O2k.

2021-02-10: Test of white PB-Sensors prototypes measuring photosynthesis in Chlamydomonas reinhardtii

2021-02-09: Start of the last biological replicates measuring oxygen kinetics of the contribution to total dark respiration of the AOX-Q branch in Chlamydomonas reinhardtii.

2021-01-29: Deliverable 2.1 submitted.

2021-01-29: MitoFit preprint publication: Magnesium Green for fluorometric measurement of ATP production does not interfere with mitochondrial respiration

2020-12-04: Start of the development of a white PB-Sensor for the PB-Module in collaboration with the algae KOLs

2020-11-06: SUIT protocols (SUIT-030 family) to determine the maximal photosynthesis rate under light- and inorganic carbon-saturating conditions optimized for microalgal living cells.

2020-10-27: First tests of the NADH-Module integrated in the NextGen-O2k.

2020-10-22: Optimization of the optimal conditions for measuring photosynthesis in the marine microalgae Phaeodactylum tricornutum with the PB-Module.

2020-10-08: Laurent Beck started with measurements on Chlamydomonas reinhardtii using the PB- and Q-Modules , supervised by PI Maria Huete-Ortega and Thomas Roach (University of Innsbruck, Institute of Botany)

2020-07-23: First isolation of mitochondria in microalgae trials

2020-07-01: Start of the development of protocols for permeabilizing microalgae cells

2020-04-24: First tests with pH electrodes

2020-03-11: First tests with Calcium Green

2020-02-20: Started development of protocols (and DL-Protocols) for the PB-Module, including new SUIT protocols for photosynthesis measurements.

2019-12-31: Determination of optimal Q-sensor measurement conditions for different biological samples (brain and heart isolated mitochondria from mouse, yeast)

2019-12-06: First version of the Cyclic Voltammetry Manual released: CV-Module

2019-12-02: Optimization of O2k-Procedures for isolated mitochondria preparations

2019-11-22: First version of NextGen-O2k Manual/SOP finalized: NextGen-O2k: Q-Module

2019-11-21: Updated beta version of cyclic voltammetry provided to the Key Opinion Leaders for testing

2019-11-18: Application of Q-redox state measurements in scientific projects

2019-10-22: 66 SUIT protocols for different O2k-Applications (i.e. oxygen consumption, hydrogen peroxide production, mitochondrial membrane potential, ATP production) which can be measured in living cells, permeabilized cells, isolated mitochondria, tissue homogenate and permeabilized fibers optimized and released with DatLab 7.4, see MitoPedia:_SUIT

2019-10-21: Four new videos for DatLab 7.4 software released - see: O2k-Videosupport DatLab 7.4

2019-09-24: Two new videos for mitochondrial preparations released - see: O2k-Videosupport Mitochondrial preparations

2019-09-23: First beta version of the cyclic voltammetry used by the Key Opinion Leaders during 1^st Q-Module workshop at the Oroboros O2k-Workshop IOC141

2019-09-20: First six videos for the Q-Module released - see: O2k-Videosupport Q-Module

2019-09-06: Development of two harmonized SUIT protocols (and DL-Protocols) for the Q-Module, both provided to the Key Opinion Leaders

2019-07-17: First tests of the new O2 analysis template DL7.4

2019-07: Started development of SUIT protocols for isolated mitochondria and permeabilzied cells

2019-06-18: Testing of the SUITbrowser by the participants at the Oroboros O2k-Workshop on high-resolution respirometry (HRR), IOC139

2019-06: Integration of cyclic voltammetry as a quality control step into the Q-Module

2019-06: Testing of the SUITbrowser