Oroboros Instruments O2k-FluoRespirometer Manual Download Page 1

 

 

Oroboros Instruments

 

 

High-Resolution FluoRespirometry

 

O2k-FluoRespirometer manual 

 

Mitochondrial Physiology Network 22.11(01):1-16 (2017) 

Version 01: 2017-07-21 O2k-Series H DatLab 7 

2017 Oroboros 

 

wiki.oroboros.at/index.php/MiPNet22.11_O2k-FluoRespirometer_manual  

 

 

 

O2k-FluoRespirometer 
manual 

 

Gnaiger E

1,2

, Sigl R

1

, Mészáros A

1

,  

Capek O

1

, Gradl L

3

, Gradl P

4

 

 

Oroboros Instruments 

Schöpfstr 18, A-6020 Innsbruck, Austria 

Email: 

[email protected]

 

www.oroboros.at

 

Mitochondria and Cell Research

 

 

 

 

 

 

Section 

1

.

  Unpack the O2k .......................................... 3 

Page 

2

.  Oroboros USB-flash drive and DatLab ............ 3 

3.

  Electric and lab space requirements ............... 4 

4.

  Start the O2k .............................................. 6 

5.

  Assembly of the O2k .................................... 6 

6.

  Start DatLab ............................................... 9 

7.

  O2k control ............................................... 14 

 

Supplement A: Videosupport ............................. S1

 

Supplement B: O2k-Chamber assembly .............. S2

 

Supplement C: O2k-Chamber volume calibration . S3

 

 

 

 

Overview

  

 

The 

Oroboros  O2k-FluoRespirometer

  provides  a  unique  high-resolution 

approach to the monitoring of cellular and mitochondrial respiratory function. 

High-Resolution FluoRespirometry

 

(HRFR) is indispensable for application of 

SUIT protocols, at low respiratory activities, kinetic transitions, and controlled 

oxygen levels. High-resolution is required for analysis of pathological effects 
causing  reduced  respiration  (mitochondrial  and  metabolic  diseases, 

ischemia-reperfusion  injury,  oxidative  stress,  apoptosis,  aging).  HRFR  is 
unique  for  simultaneous  measurement  of  respiration  and  fluorometric 

signals 

(H

2

O

2

 

flux, 

mt-membrane 

potential, 

Ca

2+

ADP-ATP 

phosphorylation), for analysis of biopsies with limited amounts of sample 

(diagnosis  of  genetic  and  acquired  mitochondrial  defects  in  pediatric 
patients, diabetes); cell cultures with limited cell numbers or low activities; 

mutants with diminished respiratory capacity; chemical oxidation rates and 
antioxidant  capacities  and  oxygen  kinetic  measurements  at  low, 
physiological intracellular oxygen levels. 

O2k-FluoRespirometer 

Summary of Contents for O2k-FluoRespirometer

Page 1: ...meter provides a unique high resolution approach to the monitoring of cellular and mitochondrial respiratory function High Resolution FluoRespirometry HRFR is indispensable for application of SUIT pro...

Page 2: ...y step instructions integrated as DL Protocols into DatLab 7 Numerous new O2k Publications on applications of the Oroboros O2k focus on critical issues of mitochondrial physiology While the Oroboros O...

Page 3: ...Titration Set 1 Unpack the O2k FluoRespirometer Box 1 or Peli Case contains the O2k Main Unit with the O2k Chambers in a finally mounted position the OroboPOS Connectors without polarographic oxygen...

Page 4: ...ates this icon on your desktop Oroboros DatLab for control of the O2k TIP2k data acquisition analysis Running DL7 For data safety reasons and optimal DatLab performance it is strongly recommend to ope...

Page 5: ...e for the ISS O2k Titration Set and handling For maximum signal stability and optimal temperature control the O2k should be placed in a position in which it is not exposed to direct sunlight and sudde...

Page 6: ...ct to electrical ground or simply touch the O2k steel housing 5 1 Instrument overview The O2k FluoRespirometer is electronically equipped with all required plugs for sensors in addition to the O2 chan...

Page 7: ...it Insert the male plug of the cable 2 into the female O2 plug The red dot on the male plug 2 faces straight upwards Each OroboPOS head can be used on O2k chamber A or B Note the POS number marked on...

Page 8: ...cable to the Fluo plug Similar to the OroboPOS 2 insert the male plug of the cable into the female Fluo plug The red dot on the male plug faces straight upwards Each Smart Fluo Sensor can be used on...

Page 9: ...or chambers A and B Usually sensors should not be switched between chambers except for troubleshooting The individual sensor number is included in the protocol for generating a database of calibration...

Page 10: ...w rpm but uncoupling of the magnet occurs at very high stirring speeds causing high signal noise Both chambers may be set at an identical stirrer speed Data recording interval s can be set between 0 2...

Page 11: ...n automatically opened Save as window the default file name has the form 2017 07 21 P1 01 DLD Year Month Day P DLD P ower O2k sequential number of the experiment extension DLD for DatLab Data Select a...

Page 12: ...metric pressure continuously recorded Peltier power Operative Peltier power applied for temperature regulation of the O2k block continuously recorded Amp A and Amp B V Amperometric signal from amperom...

Page 13: ...is on Illum on highlighted in yellow or off Illum off grey Toggle by a L double click When the windows to the glass chambers are closed with optical sensors the illumination is not visible and must b...

Page 14: ...ps for renaming or deleting O2k setups 7 2 O2k control F7 with the O2k connected The O2k control key F7 provides fast access to the O2k control window Load setup After selecting a setup it must be L l...

Page 15: ...ibed above with the next sequential number After re connecting the O2k continues operation uninterrupted the experimental time is reset to zero and data recording is restarted with automatic saving to...

Page 16: ......

Page 17: ...t O2k Chamber Stirrer Bar white PVDF 15x6 mm OroboPOS Holder O2k Chamber Holder A2_Insert OroborPOS OroboPOS Connector A3_O2k Chamber volume calibration Volume Calibration Ring Stopper black PEEK coni...

Page 18: ...chamber into the copper block of the O2k FluoRespirometer 2 Screw the POS holder 1 into a position where the glass chamber with the angular cut towards the POS holder is slightly lifted upwards 3 Scre...

Page 19: ...of the stopper black PEEK is not part of the effective chamber volume When titrating 2 1 ml insert the tip of the pipette to the wall of the glass chamber to add the entire volume Liquid must not be l...

Page 20: ...the manual Trouble shooting of test versions DatLab 7 1Andras M sz ros Oroboros Instruments AM wrote and edited the manual Taking editing photos and images 1Ondrej Capek Oroboros Instruments OC was m...

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