Genetic Fluorescent Reporters
Genetic Fluorescent Reporters - Related Videos
03:04
Generation of a Fluorescent Reporter Strain in Pseudomonas aeruginosa
0 Views •
2025
Source: Rugjee, K. N. et. al., Establishment of a High-throughput Setup for Screening Small Molecules That Modulate c-di-GMP Signaling in Pseudomonas aeruginosa. J. Vis. Exp. (2016)This video demonstrates the generation of a cyclic-di-GMP-responsive reporter strain in Pseudomonas aeruginosa using electroporation. Successful transformation is confirmed through antibiotic selection and fluorescence-based...
13:14
Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications
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Cited by 4 •
2015
Since the discovery of the green fluorescent protein gene, fluorescent proteins have impacted molecular cell biology. This protocol describes how expression of distinct fluorescent proteins through genetic engineering is used for barcoding individual cells. The procedure enables tracking distinct populations in a cell mixture, which is ideal for multiplexed applications.
03:49
A Luciferase-Fluorescent Reporter Influenza Virus to Track Viral Infections
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2025
This video demonstrates a method to track viral infections in mouse models using a recombinant luciferase and fluorescence protein-expressing bi-reporter Influenza A virus. This method enables both in vivo and ex vivo studies, enabling researchers to gain insights into viral dynamics and the effects of interventions.
09:57
Imaging Membrane Potential with Two Types of Genetically Encoded Fluorescent Voltage Sensors
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Cited by 5 •
2016
A method for imaging changes in membrane potential using genetically encoded voltage indicators is described.
03:15
Screening of Cyclic di-GMP Modulators Using Bacteria Expressing a Fluorescent Reporter Protein
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2025
Source: Rugjee, K. N., et al. Establishment of a High-throughput Setup for Screening Small Molecules That Modulate c-di-GMP Signaling in Pseudomonas aeruginosa. J. Vis. Exp. (2016).This video demonstrates the screening of cyclic di-GMP modulators using genetically modified bacteria that express green fluorescent protein (GFP) under the control of a c-di-GMP-sensitive promoter. Fluorescence is measured in a multi-well plate containing different test compounds. Compounds that inhibit c-di-GMP...
FAQs
The promoter or regulatory sequence determines where and when fluorescence appears. When linked to a fluorescent protein or activity-sensitive indicator, it connects target-cell expression or a molecular change to an optical signal. This regulatory control gives the reporter biological specificity, allowing researchers to associate observed fluorescence with a selected cell population, gene, or activity-related process.
Fluorescent proteins primarily reveal expression in cells controlled by the chosen regulatory sequence, making them useful for identifying populations or tracking development. Activity-sensitive indicators instead report molecular changes associated with neural activity, such as calcium changes. The distinction determines whether fluorescence is interpreted mainly as a marker of cellular identity and gene expression or as a dynamic signal of activity.
Excitation and emission wavelengths provide the optical basis for detecting reporter signals. Researchers illuminate the engineered system at the appropriate excitation range and observe emitted fluorescence at its characteristic wavelength, while the biological meaning comes from the reporter's genetic placement or activity sensitivity. This connects an optical measurement to a defined neural population or process during live imaging.
Genetic specificity helps researchers follow the same selected neuronal population or biological process during live imaging over time. Because the reporter is linked to a chosen promoter or regulatory sequence, fluorescence can remain associated with the intended target rather than an undefined collection of cells. This supports developmental studies, circuit mapping, and longitudinal analysis of brain function.
A typical workflow begins by choosing the cell population, gene, or neural activity to monitor, then pairing the relevant promoter or regulatory sequence with a fluorescent protein or activity-sensitive indicator. After the engineered reporter is expressed, researchers use live imaging to detect fluorescence and relate its pattern or change to the selected biological target. Interpretation depends on the reporter design.
They are useful when researchers need to identify neuronal populations, trace cell development, monitor gene expression, or visualize signals associated with neural activity. Their compatibility with live imaging also supports circuit mapping, disease modeling, and longitudinal studies. The same general strategy can therefore connect cellular identity, molecular regulation, and changing brain function within living systems.
Fluorescence can reveal which cells express a selected reporter, where a target neuronal population is located, or how an activity-sensitive signal changes over time. In neuroscience, these observations may indicate developmental patterns, gene-expression states, or calcium changes associated with neural activity. Researchers can use the resulting spatial and dynamic information to study circuits and brain function.