Real-time Metabolic Analysis
Real-time Metabolic Analysis - Related Videos
07:45
Metabolic Characterization of Polarized M1 and M2 Bone Marrow-derived Macrophages Using Real-time Extracellular Flux Analysis
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Cited by 275 •
2015
Metabolic reprogramming is a characteristic and prerequisite for M1 and M2 macrophage polarization. This manuscript describes an assay for the measurement of fundamental parameters of glycolysis and mitochondrial function in mouse bone marrow-derived macrophages. This tool can be applied to investigate how particular factors affect the macrophage’s metabolism and phenotype.
08:53
Real Time Analysis of Metabolic Profile in Ex Vivo Mouse Intestinal Crypt Organoid Cultures
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Cited by 18 •
2014
Small intestinal crypt organoids cultured ex vivo provide a tissue culture system that recapitulates growth of crypts dependent on stem cells and their niche. We established a method to assay the metabolic profile in real time in primary mouse crypt organoids. We found organoids maintain physiological properties defined by their source.
04:41
Real-Time Monitoring of Energy Metabolism in Human NK Cells Using an Extracellular Flux Analyzer
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2025
This video demonstrates the method to analyze energy metabolism in activated human NK cells using an extracellular flux analyzer. Different inhibitors of the electron transport chain complexes impact ATP synthesis, which is measured through the oxygen consumption rate or OCR, providing insights into cellular metabolism and bioenergetics.
05:41
Metabolic Profile Analysis of Zebrafish Embryos
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Cited by 42 •
2013
Zebrafish represent a powerful vertebrate model that has been under-utilised for metabolic studies. Here we describe a rapid way to measure the in vivo metabolic profile of developing zebrafish that allows the comparison of different mitochondrial function parameters between genetically or pharmacologically manipulated embryos, thereby increasing the applicability of this organism.
09:05
Real-Time Metabolic Detection in Living Cells Using Hyperpolarized 13C NMR
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Cited by 1 •
2025
This protocol integrates a custom-built bioreactor system with hyperpolarized ¹³C NMR spectroscopy to measure real-time pyruvate metabolism in live cells, improving reproducibility and kinetic accuracy for metabolic studies and drug screening applications.
FAQs
Measurements of oxygen consumption and extracellular acidification provide complementary readouts of changing cellular metabolism. Oxygen consumption reports activity associated with oxidative phosphorylation, whereas extracellular acidification helps reveal glycolytic activity. Tracking both signals over time allows researchers to identify transitions between these energy-producing modes and determine how cellular metabolism responds as experimental conditions change.
Continuous monitoring adds information that a single endpoint measurement cannot provide. By recording cells before and after a defined exposure, investigators can determine when a metabolic response begins and whether it persists, changes, or reverses. This temporal pattern supports more precise comparisons of responses to nutrients, inhibitors, pathogens, or immune stimuli.
A shift between oxidative phosphorylation and glycolysis indicates that cells have altered how they generate energy under the tested conditions. Comparing oxygen consumption with extracellular acidification helps identify that change rather than relying on one signal alone. In immunology and infection studies, the pattern can be related to cellular responses after immune stimulation or pathogen exposure.
Metabolic measurements connect changes in energy use with cellular function during immune activation or infection. Researchers can examine whether immune stimuli or pathogens alter the balance between oxidative phosphorylation and glycolysis, then compare those patterns across conditions. This approach helps characterize pathogen-driven remodeling and supports analysis of interactions between microbial activity and host-cell metabolism.
A typical workflow begins by monitoring living cells to establish their metabolic signal, followed by exposure to a selected nutrient, inhibitor, pathogen, or immune stimulus. Oxygen consumption and extracellular acidification are then tracked continuously before and after that intervention. Researchers compare the resulting time-dependent profiles to determine how the treatment changes metabolic activity.
This approach is useful when investigators need to evaluate immune-cell responses, characterize how pathogens remodel host or microbial metabolism, or examine microbial-host interactions dynamically. It can also support testing of candidate therapeutics by showing how treatment changes metabolic activity under the selected experimental conditions, rather than assessing only a final cellular measurement.
The method can show whether a candidate therapeutic changes oxygen consumption, extracellular acidification, or the balance between oxidative phosphorylation and glycolysis. Because measurements are collected as conditions change, investigators can assess the timing and direction of the metabolic response. Linking these profiles to cellular function helps determine how treatment affects immune or infection-related processes.