Genetic Fluorescent Markers
Genetic Fluorescent Markers - Related Videos
08:45
Duplex Digital PCR for Simultaneous Quantification of Dual Genetic Markers
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2025
In this video, we demonstrate the duplex digital PCR (ddPCR) technique — a modification of the traditional PCR technique that is useful in detecting two different genetic markers simultaneously. A single PCR reaction is partitioned into nanoliter-sized emulsified droplets that are independently amplified, and the detection of differently colored fluorescence amplification signals from the fraction of droplets is used to compute the initial concentration of the target sequences.
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.
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.
03:19
Transduction to Label PDX Tumor Cells: Introducing Lentivirus Expressing Fluorescent Marker into the Tumor Cell In Vitro
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2023
The following video describes a technique of transduction to label (patient-derived xenografts) PDX tumor cells, which is used for introducing lentivirus expressing green-fluorescent protein and luciferase reporters into the tumor cell in vitro.
08:50
Genetic Screens
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2023
Genetic screens are critical tools for defining gene function and understanding gene interactions. Screens typically involve mutating genes and then assessing the affected organisms for phenotypes of interest. The process can be “forward”, where mutations are generated randomly to identify unknown genes responsible for the phenotypes, or it can be “reverse”, where specific genes are targeted for mutation to observe what phenotypes are produced.Here, JoVE reviews various types of genetic...
FAQs
These markers are read by illuminating the sample at a suitable excitation wavelength and detecting the resulting visible fluorescence. The emitted signal provides a measurable location or intensity associated with the labeled cell, protein, or pathogen. This optical readout allows researchers to follow biological events in living or cultured material without chemically staining every sample.
Because a promoter can control the reporter gene, fluorescence becomes associated with activity of the linked genetic program rather than simply with the presence of a tagged structure. Researchers can therefore use the signal to visualize where or when that activity is represented in cells or pathogens. This differs from using a fluorescent protein fusion to track a molecule directly.
A fluorescent protein fusion attaches the fluorescent output to a selected protein or biological entity, supporting direct tracking of that labeled target. A promoter-controlled reporter instead connects fluorescence with gene activity. The choice determines whether the experiment emphasizes movement or interaction of a labeled component, or the spatial and temporal pattern of a genetic program.
Selection should follow the biological question. A fusion is suited to following a labeled protein or pathogen, whereas promoter control is suited to reporting gene activity. Researchers also need to decide whether the output will be observed in cultured cells or tissues and whether the experiment requires live-cell imaging, cell sorting, or quantitative analysis.
Researchers first choose a DNA-encoded label that matches the target or activity of interest, then examine the resulting fluorescence under suitable excitation. They can image labeled cells, proteins, or pathogens in cultured cells or tissues, and subsequently use the signal for cell sorting or quantitative analysis. The workflow connects marker design to a measurable biological outcome.
In immunology, fluorescence provides a way to visualize immune-cell trafficking within cultured cells or tissues. Researchers can follow where labeled immune cells appear and compare their behavior with other marked components during host-pathogen studies. Because the signal is measurable, the same experiments can support both visual observation and quantitative analysis of immune-cell movement.
Marking a pathogen or a relevant host component allows investigators to observe infection-associated events such as pathogen entry, replication, and interactions with host cells. Fluorescence can be followed during imaging rather than relying only on chemical staining. This makes the markers useful for examining host-pathogen dynamics in cultured cells or tissues.
These signals can serve as inputs for cell sorting as well as imaging. Sorting separates cells according to their detectable fluorescence, while quantitative analysis uses measured signal to examine host-pathogen dynamics. Consequently, the approach supports more than visual localization: it can organize cell populations and provide numerical evidence about biological interactions observed in the experiment.