Bioluminescence Assay
Bioluminescence Assay - Related Videos
08:32
The ATP Bioluminescence Assay
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2023
In fireflies, the luciferase enzyme converts a compound called luciferin into oxyluciferin, and produces light or “luminescence” as a result. This reaction requires energy derived from ATP in order to proceed, so researchers have exploited the luciferase-luciferin interaction to gauge ATP levels in cells. Given ATP’s role as the cell’s currency of energy, the ATP bioluminescence assay can provide insight into cellular metabolism and overall cell health.In this video, JoVE discusses cellular...
02:37
In Vivo Assay to Correlate Bacterial Bioluminescence with Cell Density
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2025
Source: Brodl, E., et al. In Situ Measurement and Correlation of Cell Density and Light Emission of Bioluminescent Bacteria. J. Vis. Exp. (2018).This video demonstrates the measurement and correlation of bioluminescence with cell density in transformed bacteria. A single bacterial colony is used to initiate a liquid culture. The culture is placed in a well at the desired cell concentration. An antibiotic and inducer are added to express the gene required for bacterial bioluminescence. Readings...
11:43
A Screenable In Vivo Assay for Mitochondrial Modulators Using Transgenic Bioluminescent Caenorhabditis elegans
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Cited by 20 •
2015
A protocol is described for in vivo detection of effects of mitochondrial inhibitors in the model organism Caenorhabditis elegans and for identification of potential enhancing compounds. This protocol can be used to screen drug libraries for compounds modulating mitochondrial function.
11:56
In Vitro Bioluminescence Assay to Characterize Circadian Rhythm in Mammary Epithelial Cells
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Cited by 12 •
2017
An in vitro bioluminescence assay to determine cellular circadian rhythm in mammary epithelial cells is presented. This method utilizes mammalian cell reporter plasmids expressing destabilized luciferase under the control of the PERIOD 2 gene promoter. It can be adapted to other cell types to evaluate organ-specific effects on circadian rhythm.
03:17
Cellular Bioluminescence Based Assay: A High Throughput Method for Combinatorial Drug Screening
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2023
In this video, we describe a method highlighting the utility of bioluminescence assay for sensitive and rapid screening of drugs. These drugs can be tested alone or in combination to assess the synergism in their mode of action.
FAQs
The luciferase reaction depends on the presence of its substrate, such as luciferin, and required cofactors. These components enable substrate oxidation and photon emission, so their availability directly supports measurable light production. In practice, the assay signal is meaningful only when the reaction conditions allow luciferase activity to proceed, making substrate and cofactor handling central to quantitative measurements.
Light intensity represents the amount of biological target linked to the luciferase reaction, rather than a single universal measurement. Depending on assay design, the signal can correspond to enzyme amount, viable cells, ATP, or another target. Researchers therefore interpret photon output according to the biological component being measured, such as pathogen abundance or host-cell viability.
The same light-producing principle can report different biological states because luciferase may be linked to distinct targets. A viability-focused assay can indicate living host cells, whereas a reporter-expressing microbe can provide information about infection burden. Identifying the target prevents researchers from treating every increase or decrease in signal as the same biological outcome.
A typical workflow connects the biological target to luciferase activity, supplies luciferin or another appropriate substrate with required cofactors, and records the emitted photons using a luminometer. The resulting signal is then related to the selected target, such as viable cells or a reporter-expressing pathogen. This sequence supports rapid quantitative measurements in cell cultures or animal models.
In infection research, microbes engineered or selected to express a luciferase reporter can produce a signal associated with their presence. Measuring emitted light provides a way to monitor pathogen burden in cell cultures and animal models. Changes in signal can then support evaluation of infection progression or the effects of treatments that alter microbial activity or abundance.
Bioluminescence assays can compare signal levels under different immune or treatment conditions. A reporter-expressing microbe can reveal immune-mediated inhibition, while measurements linked to host-cell viability can indicate cellular effects during infection or drug exposure. This approach gives researchers a rapid quantitative readout for assessing pathogen control, host-cell outcomes, and drug activity.