Respirometric Assay
Respirometric Assay - Related Videos
08:11
Isolation of Mitochondria from Mouse Skeletal Muscle for Respirometric Assays
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Cited by 2 •
2022
Here, we describe a detailed method for mitochondria isolation from mouse skeletal muscle and the subsequent analysis of respiration by Oxygen Consumption Rate (OCR) using microplate-based respirometric assays. This pipeline can be applied to study the effects of multiple environmental or genetic interventions on mitochondrial metabolism.
08:03
Unveiling Xenobiotic Transport and Effects in Isolated Mitochondria: Insights from Respirometric and Enzymatic Assays
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Cited by 1 •
2025
This study introduces a methodology for studying xenobiotic effects on mitochondrial function via respirometric and enzymatic assays. It examines electron transport chain modulation in various species, offering insights for targeted biopesticide development against insect vector-borne diseases.
11:10
Respirometric Oxidative Phosphorylation Assessment in Saponin-permeabilized Cardiac Fibers
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Cited by 32 •
2011
Saponin-permeabilized fiber preparation in conjunction with respirometric oxidative phosphorylation analysis provides integrative assessment of mitochondrial function. Mitochondrial respiration in physiological and pathological states can reflect various regulatory influences including mitochondrial interactions, morphology and biochemistry.
11:27
Preparation and Respirometric Assessment of Mitochondria Isolated from Skeletal Muscle Tissue Obtained by Percutaneous Needle Biopsy
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Cited by 24 •
2015
Methods for biopsy of Vastus lateralis, preparation of purified mitochondria, and respirometric profiling are described. The use of small muscle volume makes this technique suitable for clinical research applications.
08:12
The TUNEL Assay
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2023
One of the hallmarks of apoptosis is the nuclear DNA fragmentation by nucleases. These enzymes are activated by caspases, the family of proteins that execute the cell death program. TUNEL assay is a method that takes advantage of this feature to detect apoptotic cells. In this assay, an enzyme called terminal deoxynucleotidyl transferase catalyzes the addition of dUTP nucleotides to the free 3’ ends of fragmented DNA. By using dUTPs that are labeled with chemical tags that can produce...
FAQs
Each signal provides a different measurement route for following respiration. Oxygen consumption reflects removal of oxygen from the assay chamber, whereas carbon dioxide production reflects gas released during substrate oxidation. Pressure changes provide an indirect signal of gas exchange. Tracking any of these variables over time allows researchers to calculate or compare respiratory activity under defined experimental conditions.
Time-resolved measurements show how respiratory activity changes during an experiment rather than providing only a single endpoint. This helps reveal differences in metabolic rate, substrate utilization, or responses to inhibitors and environmental conditions. The resulting trajectory can connect a biochemical change with its timing, making it useful for comparing dynamic respiratory behavior among samples.
Substrates provide oxidizable inputs whose use can be related to respiratory activity, while inhibitors test how respiration responds when a biochemical process is disrupted. Comparing gas exchange before and after these experimental changes can indicate whether metabolism depends on a particular substrate or pathway. This approach links measured respiratory behavior with underlying energy metabolism.
The biological sample is placed in a controlled chamber so that gas exchange can be monitored while experimental conditions are changed deliberately. Depending on the research question, the system may contain cells, microorganisms, mitochondria, or enzymes and may be tested with different substrates, inhibitors, or environmental conditions. Consistent control makes changes in respiratory activity easier to compare.
A general workflow places the selected biological material in the assay chamber, establishes the chosen experimental condition, and records oxygen consumption, carbon dioxide production, or pressure over time. Researchers then compare the resulting respiratory measurements across samples or treatments. This sequence connects the biological preparation and experimental change to quantitative metabolic outcomes.
The method is useful when researchers need quantitative information about energy metabolism in cells, microorganisms, mitochondria, or enzyme systems. Applications include evaluating metabolic rates, examining mitochondrial function, measuring microbial growth, and studying substrate utilization. It also supports tests of how respiratory pathways respond to inhibitors or environmental changes, providing biochemical context for observed activity.