Gfp Signal Detection
Gfp Signal Detection - Related Videos
02:42
RNA CISH Signal Detection: A Technique to Detect Chromogenic Signals During RNA In Situ Hybridization in Intact Tissue Specimens
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2023
This video presents the protocol for signal detection during RNA chromogenic in situ hybridization. This technique can be used to diagnose an active oncogenic infection visually in an intact tissue specimen.
03:24
Establishing Experimental Metastases Mouse Model: Implanting GFP Expressing CRC Organoid Cells in PDX Model to Detect Micrometastases
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2023
This video describes the protocol for generating metastases by the PDX derived CRC organoids labeled with GFP lentivirus into the spleen of an immunodeficient mouse. Cancer micrometastases colonizing other organs can be detected using a fluorescence microscope to assess the GFP expression.
07:59
Tracking Cells in GFP-transgenic Zebrafish Using the Photoconvertible PSmOrange System
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Cited by 2 •
2016
We established the photoconvertible PSmOrange system as a powerful, straight-forward and cost inexpensive tool for in vivo cell tracking in GFP transgenic backgrounds. This protocol describes its application in the zebrafish model system.
03:38
Measuring Mycobacterium smegmatis Mistranslation Rates Using a GFP/Luciferase Reporter System
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2025
Source: Chen, Y., et al., Measurement of Specific Mycobacterial Mistranslation Rates with Gain-of-function Reporter Systems. J. Vis. Exp. (2019)This video demonstrates a method to quantify mistranslation in Mycobacterium smegmatis using an inducible GFP and secreted mutant luciferase reporter system. It shows how luminescence resulting from mistranslation is measured and compared to GFP fluorescence to evaluate the impact of a test molecule on translational accuracy.
07:55
Imaging the Intracellular Trafficking of APP with Photoactivatable GFP
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Cited by 6 •
2015
While the transport of cell surface proteins is relatively easily studied, visualizing the trafficking of intracellular proteins is much more difficult. Here, we use constructs incorporating photoactivatable GFP and demonstrate a method to accurately follow the amyloid precursor protein from the Golgi apparatus to down-stream compartments and follow its clearance.
FAQs
Detection depends on illuminating the sample at an appropriate excitation wavelength and collecting the green light emitted at a longer wavelength. This separation allows a fluorescence microscope or related imaging system to distinguish the reporter’s signal from the incoming illumination. The resulting measurement identifies locations where matured GFP is present within cells or tissues.
The GFP chromophore must form and mature before illumination produces the measurable green fluorescence used for detection. Consequently, an observed signal reflects reporter molecules that have reached this fluorescent state, rather than simply every newly produced reporter molecule. This condition is important when relating fluorescence patterns to gene expression or protein localization during development.
The meaning of a GFP pattern depends on how the reporter is designed and what it marks. Signals can indicate gene expression, reveal where a protein is localized, or label cells for tracking. These distinctions let researchers connect molecular activity with developmental patterns, including growth, differentiation, morphogenesis, cell movements, and lineage relationships.
A basic workflow is to examine the reporter-containing cells, tissue, embryo, or model organism with illumination at an appropriate excitation wavelength, then capture the longer-wavelength green emission using fluorescence microscopy or a related imaging system. Researchers can compare the resulting signal location and timing with developmental structures or cellular behaviors to interpret the reporter pattern.
It is especially useful when researchers need to observe developmental events while they unfold in living embryos or model organisms. Repeated fluorescence measurements can show where reporter activity appears, how labeled cells move, and how tissues form. This approach links changing molecular or cellular patterns with the progression of growth, differentiation, and morphogenesis.
GFP reporter measurements can show spatial and temporal patterns associated with genes that regulate development. Researchers may use those patterns to examine when activity appears, where it occurs, and how it relates to tissue formation or cell behavior. The observations help connect gene regulation with larger developmental outcomes such as differentiation, growth, and morphogenesis.