Difference between revisions of "NextGen-O2k Scientific developments"

Latest revision as of 13:23, 3 April 2022

2022

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.

Project Year 2: During the second project year, measurement conditions and protocols were optimized for the PB- and Q-Modules with a variety of biological samples. Additionally, the NADH-Module for evaluation of the NADH-pool in physio-pathological conditions was integrated in the NextGen-O2k. Testing of modules for the DatLab8 software were conducted and improvements applied. Moreover, SUIT protocols for general, pre-clinical, and algal biotechnology applications were developed. Quality control was maintained during the second period of project, an important factor in the development of the NexGen-O2k instrument and relevant protocols.

Progress and next steps

See » Track record

Luiza Cardoza, PhD. testing the Q-Module and DatLab8
Microalgae for testing PB module

NextGen-O2k MitoFit Preprints

Link	View	Reference	Year
Donnelly 2023 MitoFit	MitoFit Preprints 2023.2. Functional hypoxia reduces mitochondrial calcium uptake.	Donnelly C, Komlódi T, Cecatto C, Cardoso LHD, Compagnion AC, Matera A, Tavernari D, Zanou N, Kayser B, Gnaiger E, Place N (2023) Functional hypoxia reduces mitochondrial calcium uptake. MitoFit Preprints 2023.2. https://doi.org/10.26124/mitofit:2023-0002 — 2024-11-17 published in Redox Biol.	2023
Went 2021 MitoFit PB	MitoFit Preprints 2021.05.	Went N, Di Marcello M, Gnaiger E (2021) Oxygen dependence of photosynthesis and light-enhanced dark respiration studied by High-Resolution PhotoRespirometry. MitoFit Preprints 2021.05. https://doi.org/10.26124/mitofit:2021-0005	2021
Huete-Ortega 2020 MitoFit Preprint Arch EA	MitoFit Preprint Arch EA20.1. High-resolution respirometry for chloroplast and mitochondrial bioenergetics in Chlamydomonas reinhardtii ― towards biotechnology exploitations	Huete-Ortega M, Di Marcello M, Iglesias-Gonzalez J, Gnaiger E (2020) High-resolution respirometry for chloroplast and mitochondrial bioenergetics in Chlamydomonas reinhardtii ― towards biotechnology exploitations. https://doi.org/10.26124/mitofit:ea20.algaeurope.0001	2020

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 ::::'''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.
+::::'''Project Year 2:''' During the second project year, measurement conditions and protocols were optimized for the PB- and Q-Modules with a variety of biological samples. Additionally, the NADH-Module for evaluation of the NADH-pool in physio-pathological conditions was integrated in the NextGen-O2k. Testing of modules for the DatLab8 software were conducted and improvements applied. Moreover, SUIT protocols for general, pre-clinical, and algal biotechnology applications were developed. Quality control was maintained during the second period of project, an important factor in the development of the NexGen-O2k instrument and relevant protocols.
 == Progress and next steps ==
-:::: '''2021-03-01:''' Changes in the firmware of the NextGen-O2k done for a 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.
+See » [[Talk:NextGen-O2k Scientific developments|Track record]]
-:::: '''2021-02-18:''' MitoFit preprint publication: [[Komlodi 2021 MitoFit Q|Coupling and pathway control of coenzyme Q redox state and respiration in isolated mitochondria]]
+<br/>
+<gallery mode=default perrow=4 widths="350px" heights="300px">
+File:Luiza Q-module testing and DatLab8.jpg|[[Cardoso Luiza HD|Luiza Cardoza, PhD.]] testing the Q-Module and DatLab8
+File:Microalgae for testing PB module.jpg|Microalgae for testing PB module
+|-
+</gallery>
+== NextGen-O2k MitoFit Preprints ==
+{{#ask:[[Instrument and method::NextGen-O2k]] [[Was published in journal::MitoFit Preprint Arch]]
+| mainlabel=Link
+|?Has info=View
-:::: '''2021-02-10:''' Test of the white PB-Sensors prototypes built by WGT measuring photosynthesis in ''Chlamydomonas reinhardtii''
+|?Has title=Reference
-:::: '''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: [[Cardoso 2021 MitoFit MgG|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
+|?Was published in year=Year
-:::: '''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.
+|format=broadtable
+|limit=5000
-:::: '''2020-10-22:''' Optimization of the optimal conditions for measuring photosynthesis in the marine microalgae ''Phaeodactylum tricornutum'' with the PB-Module.
+|offset=0
-:::: '''2020-10-08:''' Laurent Beck started with measurements on ''Chlamydomonas reinhardtii'' using the PB- and Q-Modules , supervised by [[Huete-Ortega Maria|PI Maria Huete-Ortega]] and Thomas Roach (University of Innsbruck, Institute of Botany)
+|sort=Was published in year
-:::: '''2020-07-23:''' First isolation of mitochondria in microalgae trials
+|order=descending
-:::: '''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: [[MiPNet24.16 DatLab8.0: CV-Module |CV-Module]]
-:::: '''2019-12-02:''' Optimization of O2k-Procedures for isolated mitochondria preparations
-:::: '''2019-11-22:''' First version of NextGen-O2k Manual/SOP finalized: [[MiPNet24.12 NextGen-O2k: Q-Module |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: [https://wiki.oroboros.at/index.php/O2k-Videosupport#DatLab_7.4 O2k-Videosupport DatLab 7.4]
-:::: '''2019-09-24:''' Two new videos for mitochondrial preparations released - see: [https://wiki.oroboros.at/index.php/O2k-Videosupport#O2k-Mitochondrial_preparations O2k-Videosupport Mitochondrial preparations]
-:::: '''2019-09-23:''' First beta version of the cyclic voltammetry used by the Key Opinion Leaders during 1<sup>st </sup>Q-Module workshop at the [[MiPNet24.02 IOC141 Schroecken AT |Oroboros O2k-Workshop IOC141]]
-:::: '''2019-09-20:''' First six videos for the Q-Module released - see: [https://wiki.oroboros.at/index.php/O2k-Videosupport#Q-Module_.28NextGen-O2k_-_DatLab_7.29 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 [[MiPNet24.06 Oxygen flux analysis - DatLab 7.4| 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), [[MiPNet24.01 IOC139 Schroecken AT| IOC139]]
-:::: '''2019-06:''' Integration of cyclic voltammetry as a quality control step into the Q-Module
-:::: '''2019-06:''' Testing of the [[SUITbrowser]]
-== Links and references ==