Fluorescent Markers
Fluorescent Markers - Related Videos
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.
09:57
Workflow for High-content, Individual Cell Quantification of Fluorescent Markers from Universal Microscope Data, Supported by Open Source Software
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Cited by 5 •
2014
Presented is a flexible informatics workflow enabling multiplexed image-based analysis of fluorescently labeled cells. The workflow quantifies nuclear and cytoplasmic markers and computes marker translocation between these compartments. Procedures are provided for perturbation of cells using siRNA and reliable methodology for marker detection by indirect immunofluorescence in 96-well formats.
06:05
Development of a Negative Selectable Marker for Entamoeba histolytica
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Cited by 3 •
2010
We report development of a negative selection system in E. histolytica based upon transgenic expression of a chimeric protein (FCU1) and selection with the prodrug 5-fluorocytosine. The FCU1 protein is a fusion of yeast cytosine deaminase and uracil phosphoribosyltransferase. Expression of FCU1 resulted in increased E. histolytica sensitivity towards 5-fluorocytosine.
13:28
Three-dimensional Confocal Analysis of Microglia/macrophage Markers of Polarization in Experimental Brain Injury
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Cited by 51 •
2013
A way to gain new insights into the complexity of the brain inflammatory response is presented. We describe immunofluorescence-based protocols followed by three-dimensional confocal analysis to investigate the pattern of co-expression of microglia/macrophage phenotype markers in a mouse model of focal ischemia.
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.
FAQs
A marker absorbs excitation light at a particular wavelength and then releases lower-energy light that can be detected as fluorescence. This difference allows microscopy systems to distinguish emitted signal from the illumination used to activate the marker. Selecting markers with different light responses helps researchers visualize multiple biological features within the same sample.
Target-specific attachment connects the fluorescent signal to a defined protein, nucleic acid, membrane, or other cellular component. As a result, the observed location or change in fluorescence provides information about that target rather than the entire cell. This specificity supports analysis of cell organization, molecular movement, gene expression, and signaling.
Combining markers allows different cellular components or molecular targets to be distinguished in one imaging experiment. Researchers can compare their locations, follow molecular movement, or assess changes in cellular conditions using separate fluorescence signals. This multiplex approach increases the amount of biological information obtained from a sample and contributes to more detailed cellular analysis.
A typical workflow begins by selecting a marker that can associate with the biological target of interest, followed by exposing the sample to appropriate excitation light. Fluorescence microscopy then detects the emitted signal and reveals where the target is located or how it changes. The resulting images support interpretation of cellular structures and processes.
Researchers choose these markers when they need to visualize structures or follow processes that are difficult to assess from overall sample appearance alone. They can examine cell organization, gene expression, signaling, development, or disease-related changes. Markers are especially useful when the experiment requires target-specific localization, molecular tracking, or comparison of several cellular features.
Fluorescent imaging can show the location of specific targets, distinguish cell or tissue types, track molecular movement, and reveal changes in cellular conditions. Depending on the targets selected, the same general approach can support studies ranging from intracellular organization to development and disease. Multiplex imaging further increases the detail and precision of these analyses.