PB-Module

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Oroboros Open Support

PB-Module



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MitoPedia O2k and high-resolution respirometry: Oroboros Open Support 



Description

PB-Module.jpg

The PB-Module has been developed for conducting measurements of PhotoBiology, including photosynthesis. It consists of the PB Light Source and electronic components which are an integral part of the NextGen-O2k. Measurements are recorded and evaluated with the DatLab 8 software.


Reference: Product details and purchase information

The PB-Module of the NextGen-O2k and O2k-J and higher

The PB-Module was developed for monitoring real-time the O2 flux from dark respiration to net photosynthesis and light-enhanced dark respiration. The light intensity provided by blue, red or white PBLS inserted in the window of the O2k-chambers can be set accurately by the DatLab 8 software.

Additional resources

ยป MiPNet26.11 NextGen-O2k: PB-Module
ยป Went 2021 MitoFit PB

Support

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Supported by project NextGen-O2k which has received funding from the European Unionโ€™s Horizon 2020 research and innovation programme under grant agreement No. 859770.

MitoPedia: NextGen-O2k

ยป NextGen-O2kยซ
TermAbbreviationDescription
Hydrogen peroxideH2O2
Hydrogen peroxide
Hydrogen peroxide, H2O2 or dihydrogen dioxide, is one of several reactive oxygen intermediates generally referred to as reactive oxygen species (ROS). It is formed in various enzyme-catalyzed reactions (e.g., superoxide dismutase) with the potential to damage cellular molecules and structures. H2O2 is dismutated by catalase to water and oxygen. H2O2 is produced as a signaling molecule in aerobic metabolism and passes membranes more easily compared to other ROS.
Light-enhanced dark respirationLEDRLight-enhanced dark respiration LEDR is a sharp (negative) maximum of dark respiration in plants in response to illumination, measured immediately after switching off the light. LEDR is supported by respiratory substrates produced during photosynthesis and closely reflects light-enhanced photorespiration (Xue et al 1996). Based on this assumption, the total photosynthetic oxygen flux TP is calculated as the sum of the measured net photosynthetic oxygen flux NP plus the absolute value of LEDR.
Mitochondrial membrane potentialmtMP, ฮ”ฮจp+, ฮ”elFep+ [V]The mitochondrial membrane potential difference, mtMP or ฮ”ฮจp+ = ฮ”elFep+, is the electric part of the protonmotive force, ฮ”p = ฮ”mFeH+.
ฮ”elFep+ = ฮ”mFeH+ - ฮ”dFeH+
ฮ”ฮจp+ = ฮ”p - ฮ”ยตH+ยท(zH+ยทF)-1
ฮ”ฮจp+ is the potential difference across the mitochondrial inner membrane (mtIM), expressed in the electric unit of volt [V]. Electric force of the mitochondrial membrane potential is the electric energy change per โ€˜motiveโ€™ charge or per charge moved across the transmembrane potential difference, with the number of โ€˜motiveโ€™ charges expressed in the unit coulomb [C].
NADH-Module
NADH-Module - 12700-01.jpg
The NADH-Module, is a component of the NextGen-O2k for simultaneous measurement of oxygen consumption and NAD(P)H autofluorescence. NAD(P)H autofluorescence is used to evaluate the redox state of the NAD(P)H-pool. The NADH-Module incorporates an UV light and NADH-Sensors which include a photodiode and specific filters.
NADH-Sensor
NADH sesnor 003.png
The NADH-Sensor has been developed as a part of the NADH-Module for simultaneous monitoring of oxygen consumption and NADH redox state. The NADH-Sensor is composed of a photodiode and equipped with three supergel R370 Italian blue filters (Rosco, US).
NextGen-O2k
NextGen-O2k Technical developments
Oxygen kineticsOxygen kinetics describes the dependence of respiration of isolated mitochondria or cells on oxygen partial pressure. Frequently, a strictly hyperbolic kinetics is observed, with two parameters, the oxygen pressure at half-maximum flux, p50, and maximum flux, Jmax. The p50 is in the range of 0.2 to 0.8 kPa for cytochrome c oxidase, isolated mitochondria and small cells, strongly dependent on Jmax and coupling state.
PB Light Source
PB-Sensor.png
The PhotoBiology Light Source (PBLS) has been designed as a part of the PB-Module to provide with an external source of light. This enables experiments for evaluating the production of O2 in the presence of light. The PBLS consists of one LED and one photodiode mounted on the PBLS head protected by a PMMA plastic cover. Three pairs of PBLS (white, blue, and red) are provided with the PB-Module. The light intensity can be regulated from 0 to 2750 ยตmolยทs-1ยทm-2 (red PBLS), from 0 to 3000 ยตmolยทs-1ยทm-2 (blue PBLS), from 0 to 3500 ยตmolยทs-1ยทm-2 (white PBLS). An integrated photodiode provides real-time measurement of the light intensity allowing for continuous adjustment to the desired value.
PB-Module
PB-Module.jpg
The PB-Module has been developed for conducting measurements of PhotoBiology, including photosynthesis. It consists of the PB Light Source and electronic components which are an integral part of the NextGen-O2k. Measurements are recorded and evaluated with the DatLab 8 software.
PhotoBiologyPBPhotoBiology is the science of the effect of light on biological processes. This includes photosynthesis, photochemistry, photophysics, photomorphogenesis, vision, bioluminescence, circadian rhythms and photodynamic therapy. Phototoxicity results from non-ionizing radiation (i.e. ultraviolet, visible and infrared radiation). Non-ionizing radiation is any type of electromagnetic radiation that does not carry enough energy per quantum (photon energy below 10 eV) to completely remove an electron from an atom or molecule. When photons interact with molecules, the molecules can absorb the photon energy and become excited, reacting with surrounding molecules and stimulating "photochemical" and "photophysical" changes. Respiration may be affected by light during photosynthesis or in dark respiration, with the transient response of light-enhanced dark respiration.
PhotodecompositionPDPhotodecomposition or photodegradation is the process of decay of organic material induced by increasing light intensity. Under aerobic conditions, the enhancement of photodecomposition by light intensity can be quantified by oxygen consumption in a controlled light regime.
PhotosynthesisPSPhotosynthesis is the process that converts light energy into chemical energy which is subsequently transformed to the physiological energy demand. Photosynthesis has a light-dependent and light-independent (dark) phase. In plants, algae, and cynobacteria, light energy is absorbed during the light phase by the pigment chlorophyll and used to split water and generate adenosine triphosphate (ATP) and reducing power - nicotinamide adenine dinucleotide phosphate (NADPH), with the net production of O2 as a waste product. During the dark phase ATP and NADPH are used to synthesize carbohydrates from CO2 through the metabolic pathway called Calvin-Benson cycle. Oxygenic photosynthesis is responsible for producing and maintaining the oxygen concentration of the Earthโ€™s atmosphere. In bacteria such as cyanobacteria, photosynthesis involves the plasma membrane and the cytoplasm. In eukaryotic cells (plants and algae), photosynthesis takes place in the chloroplasts.
Q-ModuleQ-Module
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The Q-Module, developed for measuring the Q redox state and cyclic voltammetry, is supported by the NextGen-O2k and consists of the Q-Sensor, integrated electronic components in the O2k, and the DatLab software.
Q-Sensor
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The Q-Sensor has been designed as a part of the Q-Module for measurements with cyclic voltammetry and voltammetry, allowing for analysis of the Q redox state. The Q-Stopper with the reference electrode is called Q-Sensor, which is plugged in the NextGen-O2k. A three-electrode system is used to detect the Q redox state. Two of the three electrodes (glassy carbon and platinum electrode) are built into the Q-Stopper, while the reference electrode is removable (Reference-Electrode\2.4 mm).
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Bioblast links: Q - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>
Coenzyme Q
ยป Coenzyme Q
ยป Quinone, Ubiquinone Q; oxidized
ยป Quinol, Ubiquinol QH2; reduced
ยป Semiquinone
ยป Coenzyme Q2
ยป Q-redox state
ยป Q-pools
Mitochondrial pathways, respiratory Complexes, and Q
ยป Q-cycle
ยป Q-junction
ยป Convergent electron flow
ยป NS-pathway
ยป FNS
ยป FNSGp
ยป N-pathway
ยป Reverse electron flow from CII to CI
ยป CI
ยป Rotenone
ยป Amytal
ยป Piericidin
ยป S-pathway
ยป CII
ยป Malonate
ยป F-pathway
ยป CETF, Electron-transferring flavoprotein complex
ยป Gp-pathway
ยป CGpDH, Glycerophosphate dehydrogenase complex
ยป CIII
ยป Myxothiazol
ยป Choline dehydrogenase
ยป Dihydro-orotate dehydrogenase
NextGen-O2k and Q-Module
ยป NextGen-O2k
ยป Q-Module
ยป Q-Sensor
ยป Cyclic voltammetry
ยป Three-electrode system
General
ยป Categories of SUIT protocols
ยป Electron transfer pathway
ยป Electron-transfer-pathway state
ยป F-junction
ยป N-junction



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Bioblast links: NADH - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>


Mitochondrial NADH and respiration

ยป Nicotinamide adenine dinucleotide
ยป NADH electron transfer-pathway state
ยป N-junction
ยป Complex I
ยป Rotenone
ยป Amytal
ยป Piericidin

Mitochondrial NADH production

ยป Tricarboxylic acid cycle
ยป Fatty acid oxidation

NextGen-O2k

ยป NextGen-O2k




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Bioblast links: Hydrogen peroxide - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>



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Bioblast links: ATP production - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>



Phosphorylation pathway

ยป Phosphorylation pathway
ยป ATP synthase
ยป Adenine nucleotide translocase
ยป Phosphate carrier
Phosphorylation pathway substrates
ยป ADP
ยป ATP
ยป Inorganic phosphate
Phosphorylation pathway inhibitors
ยป Oligomycin
ยป Carboxyatractyloside
ยป Atractyloside
ยป Bongkrekik acid

Coupling control

ยป Oxidative phosphorylation
ยป OXPHOS capacity
ยป P-L control efficiency
ยป ROUTINE respiration
ยป R-L control efficiency

Respiratory complexes and coupling

ยป Complex I
ยป Complex III
ยป Complex IV
ยป Proton pump
ยป Electron-transfer-pathway state

ATP production measurement

ยป Magnesium Green



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Bioblast links: pH and protons - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>
pH and protons
ยป pH
ยป hydrogen ion H+
ยป hydron H+
ยป hydronium ion H3O+
ยป hydride H-
ยป proton p+
ยป pH buffering capacity
ยป proton flux
ยป proton pump versus hydrogen ion pump
ยป proton leak
ยป proton slip
ยป protonmotive force
Oroboros pH
ยป Oroboros Catalogue: O2k-pH ISE-Module
ยป Oroboros Manual pH electrode: MiPNet23.15 O2k-pH ISE-Module
ยป MiPNet08.16 pH calibration
ยป File:PH-Calibration-List.xls
ยป NextGen-O2k, ratiometric: Carboxy SNARF 1
ยป NextGen-O2k, ratiometric: HPTS
ยป pH calibration buffers
Oroboros Publications
ยป O2k-Publications: O2k-pH ISE-Module
HRR - general
ยป Oroboros Manual: MiPNet22.11 O2k-FluoRespirometer manual
ยป O2k signals and output
ยป MiPNet14.06 Instrumental O2 background
ยป MiPNet19.18A O2k-Series G: Start
ยป ESD
ยป O2k configuration
ยป O2k control
ยป Oroboros FluoRespirometer
ยป Oroboros Main Unit#O2k-Series
ยป Titration-Injection microPump
ยป Compare: O2k-TPP+_ISE-Module
DatLab
ยป DatLab 7
ยป MiPNet26.06 DatLab 7: Guide
ยป DatLab 6
ยป MiPNet19.18C DatLab 6: Guide
ยป MiPNet19.18D O2k-Series G and DatLab 6: Calibration
ยป Reference layouts for DatLab graphs



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Bioblast links: Calcium - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>



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Bioblast links: PhotoBiology and plant physiology - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>

PhotoBiology: photosynthesis

ยป PhotoBiology
ยป Photosynthesis

Plant physiology: respiration

ยป Photorespiration
ยป Chlororespiration
ยป Light-enhanced dark respiration

NextGen-O2k and PB-Module

ยป NextGen-O2k
ยป PB-Module
ยป PB-Sensor



MitoPedia methods: Respirometry 


MitoPedia O2k and high-resolution respirometry: Oroboros hardware, O2k-Respirometry 


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MitoPedia:NextGen-O2k 


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