High Throughput Respirometry
High Throughput Respirometry - Related Videos
10:25
Isolation of Intact Mitochondria from Skeletal Muscle by Differential Centrifugation for High-resolution Respirometry Measurements
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Cited by 26 •
2017
Here, a quadriceps muscle specimen is taken from an anaesthetized pig and mitochondria are isolated by differential centrifugation. Then, the respiratory rates of mitochondrial respiratory chain complexes I, II and IV are determined using high-resolution respirometry.
09:44
High-throughput Screening for Chemical Modulators of Post-transcriptionally Regulated Genes
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2015
Here we describe a cell-based reporter gene assay as a valuable tool to screen chemical libraries for compounds modulating post-transcriptional control mechanisms exerted through 3’ UTR.
06:19
High-Throughput Screening to Obtain Crystal Hits for Protein Crystallography
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Cited by 16 •
2023
This protocol details high-throughput crystallization screening, ranging from the 1,536 microassay plate preparation to the end of a 6 week experimental time window. Details are included about the sample setup, the imaging obtained, and how users can perform analyses using an artificial intelligence-enabled graphical user interface to quickly and efficiently identify macromolecular crystallization conditions.
10:08
High-Resolution Respirometry in a Small-Volume Chamber
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Cited by 2 •
2025
Here, we show that the 0.5-mL chamber for high-resolution respirometry shows results that are consistent with the 2.0-mL chamber if appropriate instrumental O2 background correction is applied. It requires less sample, which is advantageous for studies with limited availability or low respiratory capacities.
04:31
Measurement of Metabolic Rate in Drosophila using Respirometry
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Cited by 49 •
2014
Metabolic disorders are among one of the most common diseases in humans. The genetically tractable model organism D. melanogaster can be used to identify novel genes that regulate metabolism. This paper describes a relatively simple method which allows studying the metabolic rate in flies by measuring their CO2 production.
FAQs
The combination of oxygen and extracellular acidification provides complementary metabolic information. A change in oxygen concentration reflects respiratory activity, whereas extracellular acidification provides a signal associated with glycolytic activity. Examining both measurements under the same experimental condition helps determine whether a response is primarily linked to mitochondrial respiration, glycolysis, or shifts involving both processes.
Sequential additions create a controlled perturbation series rather than a single endpoint measurement. Substrates can support metabolism, while inhibitors or other compounds alter the measured response. Comparing signals before and after each addition helps reveal how cells use energy and how their respiratory behavior changes when a pathway or experimental condition is modified.
The method can apply the same measurement sequence to immune cells, infected cultures, and pathogen-exposed samples across many wells. Differences in oxygen use or extracellular acidification can then be related to immune activation or infection-associated metabolic shifts. This comparative design evaluates multiple biological conditions in parallel under controlled experimental settings.
Controlled conditions help distinguish changes caused by the biological state of a sample from changes produced by an added compound. Keeping the measurement environment and addition sequence consistent supports comparisons among activated, infected, or drug-exposed samples. The resulting profiles can reveal altered bioenergetic pathways rather than merely showing that overall metabolic activity differs.
A microplate workflow begins by placing multiple cell or culture samples into parallel assay wells, then monitoring oxygen concentration and, where included, extracellular acidification. Substrates, inhibitors, or other compounds are added sequentially while sensors track changing signals. The resulting time-dependent measurements support direct comparison of respiratory behavior across the tested samples.
Key components include a microplate format, sensors for oxygen concentration, and compounds that perturb metabolism. Substrates probe how cells generate energy, whereas inhibitors or other compounds help distinguish contributions from mitochondrial respiration and glycolysis. Including extracellular-acidification measurements adds a second metabolic signal, strengthening interpretation of how energy use changes across conditions.
In immunology and infection research, the approach can compare immune cells with infected cultures or pathogen-exposed samples. These comparisons help investigators examine metabolic changes associated with immune activation and host-pathogen interactions. The method can also test how drug responses differ between biological conditions, supporting studies of cellular metabolism during infection and immune challenge.
Measurements can produce metabolic signatures that characterize altered bioenergetic pathways rather than only a single respiration value. Researchers may use these profiles to compare disease-related samples, evaluate responses to compounds, and investigate patterns linked to disease progression. Interpretation requires considering oxygen and acidification signals together with the experimental additions that generated each response.