Description
Reference: MiPNet??
»
MitoPedia O2k and high-resolution respirometry:
O2k hardware
MitoPedia O2k and high-resolution respirometry:
O2k-Open Support
O2k-Fluo Smart-Module
O2k-FluoRespirometer
The Oroboros O2k-FluoRespirometer (O2k-Series H) - the experimental system complete for high-resolution respirometry (HRR), including fluorometry, the TIP2k and the O2k-sV-Module allowing simultaneous monitoring of oxygen consumption together with either ROS production (AmR), mt-membrane potential (TMRM, Safranin and Rhodamine 123), Ca2+ (CaG) or ATP production (MgG).
The O2k-FluoRespirometer supports all add-on O2k-Modules: O2k-TPP+ ISE-Module, O2k-pH ISE-Module, O2k-NO Amp-Module, enabling measurement of mt-membrane potential with ion sensitive electrodes (ISE for TPP+ or TPMP+ ) or pH.
O2k-Series H: innovations |
Oroboros (2017-07-11) Mitochondr Physiol Network
Abstract: Mészáros A, Krumschnabel G, Gradl P, Schwaninger H, Gnaiger E (2017) O2k-Series H: innovations. Mitochondr Physiol Network 22.09 O2k-Series H is the latest version of the Oroboros O2k for high-resolution respirometry and O2k-MultiSensor applications. The O2k-Series H sets a new standard in merging high-end technology and corporate social responsibility.
• O2k-Network Lab: AT_Innsbruck_Oroboros
Useful links:
Select the Smart Fluo-Sensors
- Switching between different excitation wavelengths and filters is achieved by simply exchanging the Smart Fluo-Sensors. Two types of optical sensors are supplied with different LEDs for fluorescence excitation, and the effective spectra of the LEDs are modified by filters.
- Select the Fluo-Sensor and filter set from section 6 of application-specific settings. Each Fluo-Sensor is delivered with a mounted filter set.
Smart Fluo-Sensor Green
- Excitation LED 525 nm (dominant wavelength) with short pass filter, emission (red) with long pass filter, individual sensors are calibrated with sensor-specific memory and direct input into DatLab 7 to obtain reproducible light intensities with different sensors, including photodiode, Filter-Cap equipped with Filter Set AmR for Amplex UltraRed and TMRM measurements when delivered.
- Two units of this item are standard components of the O2k-FluoRespirometer and the O2k-Fluo Smart-Module. Smart Fluo-Sensors can be used with O2k-Series H and higher.
Smart Fluo-Sensor Blue
- Excitation LED 465 nm (dominant wavelenght) with short pass filter, emission (red) with long pass filter, individual sensors are calibrated with sensor-specific memory and direct input into DatLab 7 to obtain reproducible light intensities with different sensors, including photodiode, Filter-Cap equipped with Filter Set Saf for measurement of mitochondrial membrane potential with safranin and rhodamine 123. Filter Set MgG / CaG for Magnesium green and Calcium green measurements are included.
- Two units of this item are standard components of the O2k-FluoRespirometer and the O2k-Fluo Smart-Module. Smart Fluo-Sensors can be used with O2k-Series H and higher.
Setup and connect Smart Fluo-Sensor to O2k
- Connect the Smart Fluo-Sensor cable to the Fluo plug of the O2k, 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 O2k-Chamber A or B. The blue frame of the chamber window and the Smart Fluo-Sensor are specially designed to align in a specific rotational position with the cable extending horizontally to left (chamber A) and right (chamber B). The Smart Fluo-Sensor is carefully inserted into the window frame and rotated into final position, leaving no gap between window frame and sensor body.
- To remove the Smart Fluo-Sensor, carefully pull out the sensor body with slight back and forth rotations. Do not pull the cable.
O2k-Fluo LED2-Module
MiPNet17.05 O2k-Fluo LED2-Module
Select the Fluorescence-Sensors
- Switching between different excitation wavelengths and filters is achieved by simply exchanging the Smart Fluo-Sensors. Two types of optical sensors are supplied with different LEDs for fluorescence excitation, and the effective spectra of the LEDs are modified by filters.
- Select the Fluo-Sensor and filter set from section 6 of application-specific settings. Each Fluo-Sensor is delivered with a mounted filter set.
Fluorescence-Sensor Green
- Excitation LED 525 nm (dominant wavelength), photodiode, Filter-Cap equipped with Filter Set AmR for Amplex UltraRed measurements when delivered.
- Two units of this item are standard components of the O2k-Fluo LED2-Module.
Fluorescence-Sensor Blue
- Excitation LED 465 nm (dominant wavelength), photodiode, Filter-Cap equipped with Filter Set Saf for measurement of mitochondrial membrane potential with Safranin when delivered. Filter sets for Magnesium green® / Calcium green® measurements are included.
- Two units of this item are standard components of the O2k-Fluo LED2-Module.
Setup and connect O2k-Fluorescence LED2-Module
![Fluorescence-Control Unit lettered.jpg](/images/thumb/d/d0/Fluorescence-Control_Unit_lettered.jpg/400px-Fluorescence-Control_Unit_lettered.jpg)
- Switch off the O2k with the power switch on the rear of the O2k-Main Unit.
- Remove both blue O2k-Window Frames. Insert the O2k-Window Tool around the outer rim of the window frame and unscrew in a counter clockwise direction.
- Remove the Sensor-Guide (‘nose’) from the O2k-Front Fixation of the Fluorescence-Control Unit.
- Place the Fluorescence-Control Unit below the O2k-Chamber Block. Align the windows of the O2k-Front-Fixation with the windows of the O2k-Chamber Block, re-insert the O2k-Window Frames, and screw them finger-tight onto the O2k-Main Unit.
- Re-attach the Sensor-Guide to the O2k-Front-Fixation. (There is no need to use the fixation screw supplied with early models).
- Place the power-cables from the rear of the Fluorescence-Control Unit in the middle below the O2k-Main Unit from front to rear. Unplug the mains power cable of the O2k and plug it into the female plug of the Fluorescence-Control Unit. Insert the male plug of the Fluorescence-Control Unit into the mains socket at the rear of the O2k.
- Connect the amperometric cables attached to the side of the Fluorescence-Control Unit to the ‘Amp’ plugs (labeled "NO" in Series D-E) on the O2k-Main Unit.
- In this configuration the O2k can be used for high-resolution respirometry and fluorometry. It is not necessary to dismount the Fluorescence-Control Unit for basic HRR when a fluorescence signal is not recorded.
Power on
- Switch on the power of the O2k-Main Unit (rear).
- Press the power switch on the front panel of the Fluorescence-Control Unit. Check that the two red/green control lights (O2k-Fluorescence LED2-Module B and upwards) or the central green control light are on.
The Oxygraph Window
- From the DatLab menu choose [Oxygraph]/[O2k-Control].
- In the O2k configuration window include serial number (#) of the sensors used.
- In the O2k configuration window the LED intensity and the gain (amplification) can be set.
- Setting the LED Intensity from DatLab: O2k-Fluorescence LED2-Module Series B and higher (modules without mechanical selector switch)
- Set the desired light intensity (0 to 2000 mv) in the field "Amp Polarization Voltage [mV]".
- Click on "Send to Oxygraph" or "Connect to Oxygraph-2k" to apply the new settings.
- If any current >= 0 is set and the Fluorescence Module is switched on, the indicator light below the respective chamber on the Fluorescence-Control Unit will be green. If the current is 0 (the LED is not used) but the Fluorescence-Control_Unit is switched on, the indicator light will be red.
- Setting the LED Intensity from DatLab: O2k-Fluorescence LED2-Module Series B and higher (modules without mechanical selector switch)
- Limitations: For O2k-Series D the LED intensity must be set to the same value for both chambers.
Note: The actual current (in mA) used to drive the LED is the value set in DatLab (Amp polarization voltage) divided by 100. Previously we quoted the suggested current. To simplify the operation we now state directly the required DatLab settings.
- Setting the LED Intensity from DatLab: O2k-Fluorescence LED2-Module Series A (modules with mechanical selector switch)
- O2k-Series A of the O2k-Fluorescence LED2-Module has a mechanical selector switch for each chamber on the front side of the Fluorescence-Control_Unit
- Set the switch located on the front panel of the Fluorescence-Control_Unit to position 9 with help of the small screw driver included with the Module.
- Follow the procedure described above for Series B and higher.
- For LED2-Module Series A it is still possible to set the LED current directly at the Fluorescence-Control_Unit. However, such set function is no longer in use. The relevant instructions can be found on the discussion page in the archive.
- Conversion of the LED intensities between the Fluorescence-Sensor of the O2k-Fluo LED2-Module (O2k-Series D to G) and the Smart Fluo-Sensor of the O2k-FluoRespirometer (O2k-Series H)
- For our new flagship, the O2k-FluoRespirometer (O2k-Series H) and software DatLab 7, the fluorescent intensity (excitation) of the Smart Fluo-Sensors is set in 'Fluo intensity' units. The Smart Fluo-Sensors are pre-calibrated to guarantee similar fluorescence intensity among Smart Fluo-Sensors of the same colour, storing the sensor-specific 'gain value' in an internal memory. Technically speaking, DatLab 7 (i) reads this 'gain value' from a connected Smart Fluo-Sensor to adjust the output intensity and (ii) automatically corrects for the non-linear relationship between intensity and 'mV' to output the correct fluorescent intensity. In contrast, the fluorescent intensity (excitation) of the Fluorescence-Sensors of the O2k-Fluo LED2-Module (O2k-Series D to G) is set directly in 'mV' units. To set the same intensity for O2k-Fluo LED2-Module (O2k-Series D to G) Fluorescence-Sensor as for a O2k-FluoRespirometer (O2k-Series H) Smart Fluo-Sensor, we have created an Excel sheet that allows to calculate it.
General
Fluorescence dyes
- » Amplex UltraRed
- » Calcium green
- » Magnesium green
- » Safranin
- » TMRM
- » Rhodamine 123
Application | Sensor | Filter set | O2k output | Light intensity (polarization voltage) - Note a | Gain | Comment |
---|---|---|---|---|---|---|
Amplex UltraRed | Fluorescence-Sensor Green or Smart Fluo-Sensor Green | AmR | Type B | 100 - 500 | 1000 (at light intensity = 100) | |
TMRM | Fluorescence-Sensor Greenor Smart Fluo-Sensor Green | AmR | Type C | 200 - 500 | 1000 | at c(TMRM) = 2 µM |
Safranin | Fluorescence-Sensor Blueor Smart Fluo-Sensor Blue | Saf | Type C | 100 - 200 for c(safranin)= 2 µM; | 1000 | at c(Saf) = 2 µM, |
Magnesium green | Fluorescence-Sensor Blueor Smart Fluo-Sensor Blue | MgG / CaG | Type B | 100 - 300 | 1000 | at c(Mg Green) = 2 µM |
Calcium green | Fluorescence-Sensor Blueor Smart Fluo-Sensor Blue | MgG / CaG | Type A and C | 100 -300 | 1000 | at c(Ca Green) = 2 µM |
Rhodamine 123 | Fluorescence-Sensor Blueor Smart Fluo-Sensor Blue | Filter Set Saf | Type III | 100 -300 | 1000 | at c(Rh123) = 1 µM |
- Note a: Set the polarization voltage [mV] for the amperometric channel (Amp) in the DatLab menu [O2k-MultiSensor \ O2k Control \ Amp polarisation voltage]. Divide the polarization voltage [mV] by 100 to obtain the current [mA] through the LED. For simple operation instructions, it is sufficient to refer to the polarization settings selected in DatLab.
Stoppers
- Use only black PEEK stoppers in conjunction with fluorometric measurements. The black stoppers can be used for all HRR applications. See MiPNet22.11 O2k-FluoRespirometer manual for calibration of the O2k-chamber volume, which is identical for PEEK and PVDF stoppers.
Selecting a Filter Set
- The Filter-Cap of each sensor can be removed for application of various filter combinations on the same optical sensor.
- Selection of a Filter Set: >> Application specific settings.
- The Filter-Cap of each sensor can be removed for application of various filter combinations on the same optical sensor.
Filter Set AmR
- The set of filters for the determination of H2O2 production with Amplex UltraRed should be used together with Fluorescence-Sensor Green. The filter set consists of 6 LED filters (round) and 6 photodiode filters (rectangular). One pair of these filters is attached to Fluorescence-Sensor Green or Smart Fluo-Sensor Green when delivered as part of the O2k-Fluorescence LED2-Module or O2k-Fluo Smart-Module. A filter set is a standard component of the O2k-Fluorescence LED2-Module, O2k-Fluo Smart-Module and/or O2k-FluoRespirometer.
- Excitation spectrum obtained with the Fluorescence-Sensor Green at 1 mA current supply and equipped with the Filter Set AmR.
- In the O2k-Fluo LED2-Module dye based-absorption filters in form of polymer films are used. If required, cut-off values related to the filters (short of a full spectrum) can be used:
- AmR excitation filter (SG 370): dye based short pass filter, 50% T: 535 nm, suppressing (<5% Transmission) lambda > 595 nm
- AmR emission filter (RL2001): dye based long pass filter, 50% T: 620 nm, suppressing (<2% transmission) lambda < 590 nm
- The spectra of excitation (LED filter: R370) and emission (photodiode filter: R2001) can be found on the webpage of our supplier, ROSCO: https://emea.rosco.com/de/node/1706 (please select the filter numbers to download the specific data sheets).
Filter Set Saf
- Set of filters for the (qualitative) determination of mitochondrial membrane potential with Safranin should be used together with Fluorescence-Sensor Blue or Smart Fluo-Sensor Blue. The filter set consists of 6 LED filters (round) and 6 photodiode filters (rectangular). One pair of these filters is inserted into the Fluorescence-Sensor Blue or Smart Fluo-Sensor Blue when delivered as part of the O2k-Fluorescence LED2-Module or O2k-Fluo Smart-Module. A filter set is a standard component of the O2k-Fluorescence LED2-Module, O2k-Fluo Smart-Module and O2k-FluoRespirometer.
- Excitation spectrum obtained with the Fluorescence-Sensor Blue at 1mA current supply and equipped with the Filter Set Saf.
- In the O2k-Fluo LED2-Module dye based absorption filters in form of polymer films are used. If required, cut-off values related to the filters (short of a full spectrum) can be used:
- Saf excitation filter: (SG357): dye based short pass filter, 50% T: 470 nm, suppressing (<5% Transmission) lambda > 530 nm
- Saf emission filter: (SG19): dye based long pass filter, 50% T: 600 nm, suppressing (<1% transmission) lambda < 550 nm
- The spectra of excitation (LED filter: R357) and emission (photodiode filter: R19) can be found on the webpage of our supplier, ROSCO: https://emea.rosco.com/de/node/1706 (please select the filter numbers to download the specific data sheets).
Filter Set MgG/CaG
- The set of filters for the determination of concentrations of Mg2+ or Ca2+ with the fluorophores Magnesium green and Calcium green, respectively, should be used together with Fluorescence-Sensor Blue or Smart Fluo-Sensor Blue. The filter set consists of 6 LED filters (round) and 6 photodiode filters (rectangular). A filter set is a standard component of the O2k-Fluorescence LED2-Module, O2k-Fluo Smart-Module and O2k-FluoRespirometer.
- Excitation spectrum obtained with the Fluorescence-Sensor Blue at 1 mA current supply and equipped with the filter set for Magnesium green® / Calcium green®.
- In the O2k-Fluo LED2-Module dye based absorption filters in form of polymer films are used. If required, cut-off values related to the filters (short of a full spectrum) can be used:
- MgG/CaG excitation filter (SG59):dye based short pass filter, 50% T: not applicable (max. transmission < 50%), suppressing (<5% Transmission) lambda > 490 nm
- MgG/CaG emission filter (RL21):dye based long pass filter,50% T: 570 nm, suppressing (<1% transmission) lambda < 525 nm
- The spectra of excitation (LED filter: R59) and emission (photodiode filter: R21) can be found on the webpage of our supplier, ROSCO: https://emea.rosco.com/de/node/1706 (please select the filter numbers to download the specific data sheets).
Mounting a Filter-Cap
- Applies to: O2k-Fluo LED2-Module (O2k-Series D to G) Fluorescence Sensor and O2k-FluoRespirometer (O2k-Series H) with Smart Fluo-Sensors
- Dismounting: Pull the Filter-Cap straight from the sensor. The Filter-Cap Guide prevents rotational movements.
- Remove all filters and store them in the filter box labeled for this Filter Set.
- Insert the filters form the selected Filter Set: The round filters of each Filter Set fit to the round window of the Filter-Cap and cover the LED, the rectangular filters fit into the rectangular window of the Filter-Cap and cover the photodiode.
- Mounting: Hold sensor and filter cap in a vertical position above you. Align the Filter-Cap with the Filter-Cap Guide (small steel rod) protruding from the sensor. Press the Filter-Cap onto the sensor without rotational movements.
Control of LED-intensity
- The light intensity of the LED (LED-intensity) and the signal amplification (Gain) can be adjusted in a wide range. The table suggests initial values, which can be optimised for specific applications.
- The settings depend on the concentration of the fluorophore, which vary between different applications. Therefore, only recommendations for specific fluorophore concentrations are given. In the Amplex UltraRed assay the fluorophore is formed during the experiment.
- The recommendations apply to experiments at 37 °C. The Fluorescence intensity increases strongly at lower temperatures. Then the light intensity is reduced to avoid off-scale signals.:::: The light intensity of the LEDs is set by the current control, independent for each fluorescence sensor (O2k-Chamber A and B). The current is controlled by DatLab in a very wide range for optimization according to sample and fluorophore requirements. At higher LED-intensity the optical sensitivity is increased, i.e. the signal change per concentration change is enhanced. However, even moderately intensive light may exert negative effects: (i) Damage to the sample reducing the biological activity. (ii) Damage to fluorophores catalyzing degradation and various side reactions. Therefore, the LED-intensity should be kept as low as compatible with a smooth signal, i.e. when the resolution is just not limited by noise or disturbances. The values indicated in the table application specific settings are only suggestions to start with. It is recommended to optimize the light intensity specifically for each application.
- The light intensity of the LED (LED-intensity) and the signal amplification (Gain) can be adjusted in a wide range. The table suggests initial values, which can be optimised for specific applications.
Gain, amplification
- The gain for the Amp channel can be set in the Control Table of the Oxygraph Control window, the section “Amp” to 1, 10, 100, or 1000. The gain setting will influence the Amp raw signal recorded in Volt. See Observing the Fluorescence Signal. The amplified signal can be recorded in the range -10 to +10 V.
- The gain setting should be chosen to obtain a maximum signal well below 10 V. If the maximum observed raw signal was 9 V in an initial experiment, then the gain should be reduced to avoid "off scale" (>9.99 V). On the other hand, if the maximum recorded raw signal was considerable lower than 1 V (e.g. 0.2 V), the gain can be increased to avoid limitation of resolution by digital noise.
- Note for advanced users: At gain 1, a current of 1 nA is recorded as a voltage of 1 mV (0.001 V). At gain 100, 1 nA corresponds to 100 mV (0.1 V), as can be seen form the table below:
1
|
0.001 | 1000
|
+-10000
|
333
|
10
|
0.01 | 100
|
+-1000
|
33
|
1000
|
1 | 1
|
+-10
|
0.3
|
The fluorescence signal
- O2k signal: The O2k-Fluo LED2-Module is operated through the amperometric (Amp)-Channel of the O2k, with electric current (ampere [Amp]) as the primary signal.
- O2k output: type A, B, or C, or combinations.
Graph layout: Three plots are available in DatLab based on the recorded signal: Amp Raw Signal, Amp Calibrated, and Amp Slope. These plots can be selected from the drop-down lines and displayed with their check boxes either on the Y1 or Y2 [Graph layout / Select Plots].
- Amp Raw Signal displays the raw voltage (including amplification) as recorded by the O2k at a given gain setting.
- Amp Calibrated is the signal after calibration with the parameters set in the O2k-MultiSensor Calibration window.
- Amp slope is the time derivative of the calibrated signal, multiplied by 1000, in units [mV(conc. Unit during calibration)/s], so if the signal was calibrated in µM [nmol/ml] the unit of the slope is pmol/(s ml). To obtain the slope of the raw signal check the appropriate box in the calibration window (DatLab 5.1.0.130 and above).
- Graphs can be generated to display oxygen and fluorescence data, or several graphs can be added to display oxygen and fluorescence data separately. Layout templates are provided, which can be modified and saved as appropriate. All graph settings can be saved as user-defined layouts, see MiPNet19.01C DatLab Guide.
O2k-Fluorometry and the TIP2k
- TiP2k: Our tests indicated that the fluorometric signal is not affected by the TIP2k needle inserted into the O2k-Chamber.
O2k-Fluorometry Workshops
Troubleshooting
Defective Fluo-Sensor or O2k or O2k-LED2 Fluo-Module
- If the raw fluorescence signal is 0 V with and wihtout fluoresence dyes, it might be possible that your Fluo-Sensor is defective. How can you localize the problem?
- 1. Check that the illumination is switch off in the ´O2k control´window.
- 2. Check the settings for the Amperometric channel in the ´O2k control´ window, in the Amperometric,Amp tab. Gain for Fluo-Sensor and the Fluo intensity cannot be 0.
- 3. The Fluo-Sensors are connected to the O2k and inserted into the O2k-Chamber in the right position (see above in the 1.3 Connect Smart Fluo-Sensor to O2k section).
- 4. If all the above-mentioned steps are checked and the detected signal with your Fluo-Sensor is 0,negative or very high, in order to localise the problem, perform the following test. Add distilled water or respiration medium into the O2k-Chamber, insert the black stopper and change the settings for fluorometric measurements (see above):
- 1. run: Fluo-Sensor A in chamber A and Fluo-Sensor B in chamber B
- 2. run: Fluo-Sensor A in chamber B and Fluo-Sensor B in chamber A
- If your Fluo-Sensor A is defective, the fluorescence signal should be 0 V both in chamber A and B. In this case, the Fluo-Sensor should be shipped to us in order to repair it.
- If the O2k or the O2k-LED2 FLuo-Module are defective, 0 V or negative fluorescence signal can be observed just in one chamber with two different Fluo-Sensors (see example below). In this case, the O2k or the Fluorescence- Control Unit (Series D-G) should be shipped to us in order to repair it.
- Please contact our customer support to analyse your problem before shippment. Please always send us original DatLab files with the above-mentioned tests to evaluate the defective Fluo-Sensor performance.
- Problem: Defective O2k or O2k-LED2 Fluo-Module
- Provided by Hazirah Hassan, University Kebangsaan Malaysia, [email protected]
- Analysing these tests after changing the Fluo-Sensors between the O2k-Chambers, it is obvious that the problem is related to the O2k or O2k-LED2 FLuo-Module, since the fluorescence signal is negative and unstable in chamber B with both Fluo-Sensors. In the third figure it is visible that the fluoresnce signal is very unstable and it makes any measurment unpossible. Both the O2k and the LED2 Fluo-Module should be shipped to our electronic workshop.
- 4. If all the above-mentioned steps are checked and the detected signal with your Fluo-Sensor is 0,negative or very high, in order to localise the problem, perform the following test. Add distilled water or respiration medium into the O2k-Chamber, insert the black stopper and change the settings for fluorometric measurements (see above):
Negative fluorescence signal
- Problem: The fluorescence value for FLuoSensor in chamber B is negative, while the other FluoSensor has ~0 V. The same result occur when the O2k-LED2 Fluo-Module is disconnected from the the O2k/switched off.
- Provided by Anne Laure Charles, FR_Strasbourg_Zoll J,anne laure charles <[email protected]
- Answer: Connacting our electronic workshop, the problem might be related to wet plug of the FluoSensors. The solution is to dry it out somehow (which gives you negative signal) e.g. hair dryer and let it run connected to the O2k for a couple of days in dry and warm environment. If these treatments cannot solve the problem and you get still negative fluorescence values, you need to send us the Fluo-Sensor back and our workshop can check it and change it if it is needed.
- » References
Sort in ascending/descending order by a click on one of the small symbols in squares below. Default sorting: chronological. Empty fields appear first in ascending order.
Was published in year | Has title | Organism | Tissue;cell | |
---|---|---|---|---|
MiPNet18.10 O2k-Specifications | 2024-03-26 | O2k-specifications for respirometry and comprehensive OXPHOS analysis. | Human | Fibroblast |
Lhuissier 2024 iScience | 2024 | Lhuissier C, Desquiret-Dumas V, Girona A, Alban J, Faure J, Cassereau J, Codron P, Lenaers G, Baris OR, Gueguen N, Chevrollier A (2024) Mitochondrial F0F1-ATP synthase governs the induction of mitochondrial fission. iScience 27:109808. https://doi.org/10.1016/j.isci.2024.109808 | Mouse | Fibroblast |
Balmaceda 2024 Biochim Biophys Acta Mol Basis Dis | 2024 | Balmaceda V, Komlodi T, Szibor M, Gnaiger E, Moore AL, Fernandez-Vizarra E, Viscomi C (2024) The striking differences in the bioenergetics of brain and liver mitochondria are enhanced in mitochondrial disease. Biochim Biophys Acta Mol Basis Dis 1870:167033. https://doi.org/10.1016/j.bbadis.2024.167033 | Mouse | Nervous system Liver |
Ravasz 2024 Sci Rep | 2024 | Ravasz D, Bui D, Nazarian S, Pallag G, Karnok N, Roberts J, Marzullo BP, Tennant DA, Greenwood B, Kitayev A, Hill C, Komlódi T, Doerrier C, Cunatova K, Fernandez-Vizarra E, Gnaiger E, Kiebish Michael A, Raska A, Kolev K, Czumbel B, Narain NR, Seyfried TN, Chinopoulos C (2024) Residual Complex I activity and amphidirectional Complex II operation support glutamate catabolism through mtSLP in anoxia. Sci Rep 14:1729. https://doi.org/10.1038/s41598-024-51365-4 | Mouse | Heart Liver |
Fitzgerald 2024 J Cachexia Sarcopenia Muscle | 2024 | Fitzgerald LF, Lackey J, Moussa A, Shah SV, Castellanos AM, Khan S, Schonk M, Thome T, Salyers ZR, Jakkidi N, Kim K, Yang Q, Hepple RT, Ryan TE (2024) Chronic aryl hydrocarbon receptor activity impairs muscle mitochondrial function with tobacco smoking. https://doi.org/10.1002/jcsm.13439 | Mouse | Skeletal muscle |
Al-Sabri 2024 Sci Rep | 2024 | Al-Sabri MH, Ammar N, Korzh S, Alsehli AM, Hosseini K, Fredriksson R, Mwinyi J, Williams MJ, Boukhatmi H, Schiöth HB (2024) Fluvastatin-induced myofibrillar damage is associated with elevated ROS, and impaired fatty acid oxidation, and is preceded by mitochondrial morphological changes. https://doi.org/10.1038/s41598-024-53446-w | Drosophila | Skeletal muscle |
Donnelly 2024 Redox Biol | 2024 | Donnelly C, Komlódi T, Cecatto C, Cardoso LHD, Compagnion A-C, Matera A, Tavernari D, Campiche O, Paolicelli RC, Zanou N, Kayser B, Gnaiger E, Place N (2024) Functional hypoxia reduces mitochondrial calcium uptake. Redox Biol 71:103037. https://doi.org/10.1016/j.redox.2024.103037 | Human Mouse | Heart Skeletal muscle |
Cefis 2024 Acta Physiol (Oxf) | 2024 | Cefis M, Dargegen M, Marcangeli V, Taherkhani S, Dulac M, Leduc-Gaudet JP, Mayaki D, Hussain SNA, Gouspillou G (2024) MFN2 overexpression in skeletal muscles of young and old mice causes a mild hypertrophy without altering mitochondrial respiration and H2O2 emission. Acta Physiol (Oxf) [Epub ahead of print]. https://doi.org/10.1111/apha.14119 | Mouse | Skeletal muscle |
Taylor 2024 Pilot Feasibility Stud | 2024 | Taylor MK, Burns JM, Choi IY, Herda TJ, Lee P, Smith AN, Sullivan DK, Swerdlow RH, Wilkins HM (2024) Protocol for a single-arm, pilot trial of creatine monohydrate supplementation in patients with Alzheimer's disease. Pilot Feasibility Stud 10:42. https://doi.org/10.1186/s40814-024-01469-5 | Human | Lymphocyte Platelet |
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MiPNet24.10 H2O2 flux analysis | 2021-10-22 | Hydrogen peroxide flux analysis using Amplex UltraRed assay in MiR05-Kit with DatLab 7.4 | ||
MiPNet26.10 MgG data analysis | 2021-09-16 | Measurement of mitochondrial ATP production using Magnesium Green: DL-Protocols and data analysis with DatLab 7.4 | ||
MiPNet17.05 O2k-Fluo LED2-Module | 2021-06-22 | O2k-Fluo LED2-Module. | ||
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MiPNet24.08 Safranin Analysis Template | 2020-05-13 | Excel template for safranin data analysis. | ||
MiPNet25.14 TPP Analysis Template | 2020-##-## | Excel template for TPP data analysis. | ||
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Spinazzi 2019 Proc Natl Acad Sci U S A | 2019 | Spinazzi M, Radaelli E, Horré K, Arranz AM, Gounko NV, Agostinis P, Maia TM, Impens F, Morais VA, Lopez-Lluch G, Serneels L, Navas P, De Strooper B (2019) PARL deficiency in mouse causes Complex III defects, coenzyme Q depletion, and Leigh-like syndrome. Proc Natl Acad Sci U S A 116:277-86. 10.1073/pnas.1811938116 | Mouse | Nervous system |
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Timpani 2016 Neurotherapeutics | 2016 | Timpani CA, Trewin AJ, Stojanovska V, Robinson A, Goodman CA, Nurgali K, Betik AC, Stepto N, Hayes A, McConell GK, Rybalka E (2016) Attempting to compensate for reduced neuronal nitric oxide synthase protein with nitrate supplementation cannot overcome metabolic dysfunction but rather has detrimental effects in Dystrophin-deficient mdx muscle. Neurotherapeutics 14:429-46. | Mouse | Skeletal muscle |
Schiffer 2016 Am J Physiol Cell Physiol | 2016 | Schiffer TA, Peleli M, Sundqvist ML, Ekblom B, Lundberg JO, Weitzberg E, Larsen FJ (2016) Control of human energy expenditure by cytochrome c oxidase subunit IV-2. Am J Physiol Cell Physiol 311:C452-61. | Human | Skeletal muscle HEK |
Power 2016 Am J Physiol Heart Circ Physiol | 2016 | Power AS, Pham T, Loiselle DS, Crossman DH, Ward ML, Hickey AJ (2016) Impaired ADP channeling to mitochondria and elevated reactive oxygen species in hypertensive hearts. https://doi.org/10.1152/ajpheart.00050.2016 | Rat | Heart |
Shirey 2016 Biochemistry | 2016 | Shirey K, Stover KR, Cleary J, Hoang N, Hosler J (2016) Membrane-anchored cyclic peptides as effectors of mitochondrial oxidative phosphorylation. Biochemistry 55:2100-11. | Rat | Heart Liver |
Larsen 2016 FASEB J | 2016 | Larsen FJ, Schiffer TA, Ørtenblad N, Zinner C, Morales-Alamo D, Willis SJ, Calbet JA, Holmberg HC, Boushel R (2016) High-intensity sprint training inhibits mitochondrial respiration through aconitase inactivation. FASEB J 30:417-27. | Human | Skeletal muscle |
Du 2015 J Biol Chem | 2015 | Du J, Rountree A, Cleghorn WM, Contreras L, Lindsay KJ, Sadilek M, Gu H, Djukovic D, Raftery D, Satrustegui J, Kanow M, Chan L, Tsang SH, Sweet IR, Hurley JB (2015) Phototransduction influences metabolic flux and nucleotide metabolism in mouse retina. J Biol Chem 291:4698-710. | Mouse | Nervous system Islet cell;pancreas;thymus |
Kluckova 2015 Cell Death Dis | 2015 | Kluckova K, Sticha M, Cerny J, Mracek T, Dong L, Drahota Z, Gottlieb E, Neuzil J, Rohlena J (2015) Ubiquinone-binding site mutagenesis reveals the role of mitochondrial complex II in cell death initiation. Cell Death Dis 6:e1749. | Other mammals | Lung;gill Other cell lines Fibroblast |
Makrecka-Kuka 2015 Biomolecules | 2015 | Makrecka-Kuka M, Krumschnabel G, Gnaiger E (2015) High-resolution respirometry for simultaneous measurement of oxygen and hydrogen peroxide fluxes in permeabilized cells, tissue homogenate and isolated mitochondria. https://doi.org/10.3390/biom5031319 | Human Mouse | Heart Nervous system HEK |
Krumschnabel 2015 Methods Mol Biol | 2015 | Krumschnabel G, Fontana-Ayoub M, Sumbalova Z, Heidler J, Gauper K, Fasching M, Gnaiger E (2015) Simultaneous high-resolution measurement of mitochondrial respiration and hydrogen peroxide production. Methods Mol Biol 1264:245-61. | Mouse | Nervous system |
Elustondo 2014 Biochim Biophys Acta | 2014 | Elustondo PA, Negoda A, Kane CL, Kane DA, Pavlov EV (2014) Spermine selectively inhibits high-conductance, but not low-conductance calcium-induced permeability transition pore. Biochim Biophys Acta 1847:231–40. | Rat | Liver |
Kiss 2014 FASEB J | 2014 | Kiss G, Konrad C, Pour-Ghaz I, Mansour JJ, Németh B, Starkov AA, Adam-Vizi V, Chinopoulos C (2014) Diaphorases provide NAD+ in anoxia. FASEB J 28:1682-97. | Mouse | Liver |
Krumschnabel 2014 Methods Enzymol | 2014 | Krumschnabel G, Eigentler A, Fasching M, Gnaiger E (2014) Use of safranin for the assessment of mitochondrial membrane potential by high-resolution respirometry and fluorometry. Methods Enzymol 542:163-81. https://doi.org/10.1016/B978-0-12-416618-9.00009-1 | Mouse | Nervous system |
Chinopoulos 2014 Methods Enzymol | 2014 | Chinopoulos C, Kiss G, Kawamata H, Starkov AA (2014) Measurement of ADP-ATP exchange in relation to mitochondrial transmembrane potential and oxygen consumption. Methods Enzymol 542:333-48. doi:10.1016/B978-0-12-416618-9.00017-0 | Human | HEK |
Pham 2014 Am J Physiol | 2014 | Pham T, Loiselle D, Power A, Hickey AJ (2014) Mitochondrial inefficiencies and anoxic ATP hydrolysis capacities in diabetic rat heart. Am J Physiol 307:C499–507. | Rat | Heart |
Iftikar 2014 Thesis University of Auckland | 2014 | Iftikar FI (2014) Testing the role of heart mitochondrial stability and function in heart failure of ectotherms exposed to heat stress. Thesis University of Auckland:167pp. | Fishes | Heart |
Soltysinska 2014 PLoS One | 2014 | Soltysinska E, Bentzen BH, Barthmes M, Hattel H, Thrush AB, Harper ME, Qvortrup K, Larsen FJ, Schiffer TA, Losa-Reyna J, Straubinger J, Kniess A, Thomsen MB, Brüggemann A, Fenske S, Biel M, Ruth P, Wahl-Schott C, Boushel RC, Olesen SP, Lukowski R (2014) KCNMA1 encoded cardiac BK channels afford protection against ischemia-reperfusion injury. PLoS One 9:e103402. | Mouse | Heart Skeletal muscle |
Hunter 2014 PhD Thesis | 2014 | Hunter FW (2014) Old target; new paradigm: Determinants of sensitivity to hypoxia-directed anticancer prodrugs. PhD Thesis:1-353. | Other mammals | CHO |
Kukat 2014 PLoS Genet | 2014 | Kukat A, Dogan SA, Edgar D, Mourier A, Jacoby C, Maiti P, Mauer J, Becker C, Senft K, Wibom R, Kudin AP, Hultenby K, Flögel U, Rosenkranz S, Ricquier D, Kunz WS, Trifunovic A (2014) Loss of UCP2 attenuates mitochondrial dysfunction without altering ROS production and uncoupling activity. https://doi.org/10.1371/journal.pgen.1004385 | Mouse | Heart |
Gnaiger 2014 MitoPathways | 2014 | Gnaiger E (2014) Mitochondrial pathways and respiratory control. An introduction to OXPHOS analysis. 4th ed. Mitochondr Physiol Network 19.12. Oroboros MiPNet Publications, Innsbruck:80 pp. — see 5th edition: Gnaiger 2020 BEC MitoPathways. | Human Mouse | Heart Skeletal muscle Fibroblast |
Reilly 2013 J Exp Biol | 2013 | Reilly BD, Hickey AJ, Cramp RL, Franklin CE (2013) Decreased hydrogen peroxide production and mitochondrial respiration in skeletal muscle but not cardiac muscle of the green-striped burrowing frog, a natural model of muscle disuse. J Exp Biol 217:1087-93. | Amphibians | Heart Skeletal muscle |
Jelenik 2013 Eur Heart J | 2013 | Jelenik T, Floegel U, Phielix E, Kaul K, Nowotny P, Partke HP, Schrader J, Roden M, Szendroedi S (2013) Non-alcoholic fatty liver disease and insulin resistance are associated with increased cardiac oxidative stress in mice. Eur Heart J 34:P5045. | Mouse | Heart |
Hara 2013 Eur J Anaesthesiol | 2013 | Hara N, Karlsson M, Sjövall F, Hansson Magnus J, Elmér E, Uchino H (2013) Early brain mitochondrial dysfunction in a mouse model of sepsis: 7AP4‐9. Eur J Anaesthesiol 30,112-112. | Mouse | Nervous system |
Iftikar 2013 PLoS One | 2013 | Iftikar FI, Hickey AJ (2013) Do mitochondria limit hot fish hearts? Understanding the role of mitochondrial function with heat stress in Notolabrus celidotus. PLoS One 8:e64120. | Fishes | Heart |
Hickey 2012 J Comp Physiol B | 2012 | Hickey AJ, Renshaw GM, Speers-Roesch B, Richards JG, Wang Y, Farrell AP, Brauner CJ (2012) A radical approach to beating hypoxia: depressed free radical release from heart fibers of the hypoxia-tolerant epaulette shark (Hemiscyllum ocellatum). J Comp Physiol B 182:91-100. | Fishes | Heart |
Gnaiger 2012 Mitochondr Physiol Network Bioblast 2012 | 2012 | Gnaiger E, Meissner B, Laner V, eds (2012) Bioblast 2012: Mitochondrial Competence. Mitochondr Physiol Network 17.12. Oroboros MiPNet Publications, Innsbruck:96 pp. ISBN 978-3-9502399-5-9 | ||
Hunter 2012 Biochem Pharmacol | 2012 | Hunter FW, Wang J, Patel R, Hsu HL, Hickey AJ, Hay MP, Wilson WR (2012) Homologous recombination repair-dependent cytotoxicity of the benzotriazine di-N-oxide CEN-209: Comparison with other hypoxia-activated prodrugs. Biochem Pharmacol 83:574–85. | CHO |
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