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MiPNet23.01 IOC126 Innsbruck AT

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MiPNet23.01 IOC126 Innsbruck AT

Publications in the MiPMap
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Innsbruck AT, 2018 Jan 24-26. Oroboros advanced O2k-Workshop on mt-membrane potential: TPP+ and fluorescent dyes. IOC126.


Oroboros (2018-01-24) Mitochondr Physiol Network

Abstract:

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Oroboros advanced O2k-Workshop on mt-membrane potential: TPP+ and fluorescent dyes. Innsbruck, Austria; 2018 Jan 24-26.


O2k-Network Lab: AT_Innsbruck_Oroboros, AT_Innsbruck_MitoFit


General information

The Oroboros O2k-Workshop is an advanced course which covers an introduction to basic concepts on mitochondrial membrane potential: proton pumps, ion gradients, electrochemical potential (proton motive force) and the measurement of mt-membrane potential in biological samples (isolated mitochondria, homogenate) with potentiometric (O2k-TPP+ ISE-Module) and fluorescence method (TMRM, safranin/O2k-Fluo LED2-Module). Data analysis - from TPP+ to mt-membrane potential - constitutes a major topic.
data analysis

Feedback

4 feedback forms returned
Question Rating (average)
In how far were your interests covered in the workshop? Please rate from 1 (not covered) to 5 (all interests covered). 4.50
Balance between practical “hands-on” work and “theoretical” explanations: Please rate from 1 (too much “hands-on”) to 5 (too much theory). 2.88
Presentation of lectures: Please rate from 1 (not understandable) to 5 (very understandable). a. in terms of content: 3.50
Presentation of lectures: Please rate from 1 (not understandable) to 5 (very understandable). b. in terms of language: 4.25
Level of tutoring during the “hands-on” work: Please rate from 1 (not enough tutoring, I felt left alone) to 5 (too much tutoring, I would have liked to work more independently). 3.25
Quality of tutoring: Please rate from 1 (poor) to 5 (excellent). 4.00
Intensity of the O2k-Course: Please rate from 1 (I would have liked to do and hear more) to 5 (I felt overwhelmed). 3.50
Your overall rating of the course? Please rate from 1 (poor) to 5 (excellent). 4.00

Venue

Information on travel and venue:IOC Innsbruck.
» Hotel suggestion: Hotel Weisses Kreuz
Mention "Oroboros" when booking in order to receive a discount.

Programme

Wednesday 24th Topic Room
09:00 Get-together: Introduction of participants and their research interests. MiPArt
09:30 Erich Gnaiger: Protonmotive force and respiratory control. MitoEAGLE preprint 2017-09-21. MiPArt
10:00 DatLab 7, DL-Protocols. MiPArt
10:30 Coffee/Tea
11:00 Hands-on: Assembly and maintenance of TPP+ and reference electrodes. MiPNet15.03 O2k-MultiSensor-ISE. MitoFit ORO Lab
12:15 Lunch break
13:30 Determination of mt-membrane potential with potentiometric O2k-TPP+ ISE-Module - an introduction. MiPArt
14:30 Hands-on: Set up of the instruments with TPP+ and reference electrodes, O2 air calibration and calibration of TPP+ electrode. MitoFit ORO Lab
16:30 Coffee/Tea
17:00 Hands-on continued: TPP+ chemical background experiment. MiPNet14.05 TPP-mtMembranePotential. MitoFit ORO Lab
Thursday 25th Topic Room
09:00 Hands-on: O2 air calibration and calibration of TPP+ electrode. MitoFit ORO Lab
09:30 Introduction in planning and performing a TPP+ experiment. MitoFit ORO Lab
10:30 Coffee/Tea
11:00 Hands-on continued: TPP+ experiment with mouse brain mitochondria. MitoFit ORO Lab
12:30 Lunch break
14:00 From the TPP+ signal to mt-membrane potential - guide through the Excel templates. TPP-Matematics-MembranePotential. MitoFit ORO Lab
14:30 Analysis of TPP+ experiment. Interdependence between O2 flux and mt-membrane potential. MitoFit ORO Lab
16:00 Coffee/Tea
16:30 The four compartment model - the effect of parameters on calculated mt-membrane potential. The analysis of the model. MiPArt
17:30 Erich Gnaiger: Protonmotive force and chemiosmotic pressure: a generalization of non-ohmic flux control of the proton leak. Abstract. MiPArt
Friday 26th Topic Room
09:00 Determination of mt-membrane potential with fluorescence method (TMRM, safranin) - O2k FluoRespirometer - an introduction. MiPNet22.11 O2k-FluoRespirometer manual, MiPNet20.13 Safranin mt-membranepotential. MiPArt
10:00 Hands-on: O2 air calibration and O2k setup for fluorescence experiment, choosing fluorescence sensors. MitoFit ORO Lab
10:30 Coffee/Tea
11:00 Hands-on: Calibration of safranin/TMRM, experiment with mouse brain mitochondria. MitoFit ORO Lab
12:30 Lunch break
14:00 Analysis of TMRM/safranin experiment, comparison of TPP+ and fluorescence results. MitoFit ORO Lab
15:30 Coffee/Tea
16:00 Comparison of TPP+ and fluorescence methods for evaluation of mt-membrane potential. Examples in different protocols. MiPArt
17:00 Interdependence between O2 flux and mt-membrane potential. MiPArt
17:30 The Bioblast wiki, O2k-Network and MitoEAGLE, feedback and conclusions. MiPArt
18:00 Farewell activity MiPArt


Tutors and Lecturers

Tutors


List of participants

Participant Institution
Duicu Oana-Maria RO Timisoara Muntean DM - University of Medicine and Pharmacy
de Kok Michéle NL Leiden Lindeman JH - Leiden University Medical Center
Fernandez-Vizarra Erika UK Cambridge Zeviani M - University of Cambridge
Hunger Miriam Hunger Miriam AT Innsbruck Oroboros
Janssen Connie NL Leiden Lindeman JH - Leiden University Medical Center
Labieniec-Watala Magdalena Labieniec-Watala Magdalena PL Lodz Watala C - University of Lodz
Michalak Slawomir Michalak Slawomir PL Poznan Michalak S - Poznan University of Medical Sciences
Perez Valencia Juan Alberto BR Rio de Janeiro - National Cancer Institute
Siewiera Karolina Siewiera Karolina PL Lodz Watala C - University of Lodz
Smirnova Irina SE Stockholm Smirnova I - Stockholm University
Steffen Anja NL Leiden Lindeman JH - Leiden University Medical Center

Registration

closed

Impressions from the IOC126

Pictures taken by: Meszaros Andras


Contact

[email protected]
Oroboros Instruments
High-resolution respirometry
Schöpfstrasse 18
A-6020 Innsbruck, Austria
Tel: +43 512 566796
Fax: +43 512 566796 20
Mitochondria and cell research

IOC recommended reading

» IOC recommended reading
» MitoPedia: Respiratory states
OXPHOS ROUTINE ETS LEAK ROX

Project support

Contribution to K-Regio project MitoFit, funded by the Land Tirol within the program K-Regio of Standortagentur Tirol.
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Labels:






ORO, IOC, MitoFit, 2018, TRACT, TRANSMIT 


MitoGlobal
O2k-Workshops and MitoFit Coaching Days are listed as MitoGlobal Events.