Gfp Detection Method
Gfp Detection Method - Related Videos
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.
03:24
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.
09:33
The Tomato/GFP-FLP/FRT Method for Live Imaging of Mosaic Adult Drosophila Photoreceptor Cells
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Cited by 26 •
2013
The Tomato/GFP-FLP/FRT method involves visualizing mosaic photoreceptor cells in living Drosophila. It can be used to follow individual photoreceptor cell fates in the retina for days or weeks. This method is ideal for studies of retinal degeneration and neurodegenerative diseases or photoreceptor cell development.
04:55
CRC Organoid Cell Labeling: A Method to Generate GFP Lentivirus-transduced Colorectal Cancer Organoid Cells
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2023
This video describes the technique of generating CRC organoids transduced by GFP lentiviral particles for enabling fluorescent imaging. These GFP labeled organoids, when placed in animal models, help to study tumor invasion and metastasis.
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
The method relies on a two-stage optical process. GFP absorbs energy from blue or ultraviolet excitation light, then releases part of that energy as green emitted light. Instruments detect this emitted signal rather than the incoming illumination. The measured fluorescence therefore indicates where GFP is present and supports visualization or quantification in biological samples.
GFP fluorescence can report several related but distinct events, depending on how the reporter is used. It can indicate gene expression, show protein localization within cells, or reveal changes in cellular behavior. In immunology and infection studies, these readouts help connect fluorescence patterns with immune responses, pathogen activity, or interactions between host cells and infectious agents.
These platforms measure GFP fluorescence at different levels of biological organization. Fluorescence microscopy provides visual information about individual cells, structures, or locations. Flow cytometry measures fluorescent properties across cells, supporting cellular analysis. Fluorescence-based plate readers provide measurements from samples in wells, making them useful for quantifying biological activity across experimental conditions.
A typical workflow begins with a sample containing a GFP-tagged pathogen, immune cell, or reporter construct. The sample is then examined using blue or ultraviolet excitation, and the emitted green fluorescence is collected with microscopy, flow cytometry, or a fluorescence-based plate reader. Researchers interpret the resulting signal according to the biological activity being monitored.
In infection research, GFP-tagged pathogens can make infection and pathogen spread observable in cells or tissues. GFP-labeled immune cells or reporter constructs can also help follow cellular responses during the interaction. These measurements allow researchers to examine how infectious agents behave alongside host cells and to relate spatial or quantitative fluorescence patterns to immune activity.
GFP-based measurements can support several outcomes, including tracking infection, monitoring immune activation, assessing pathogen spread, and quantifying biological activity. Researchers may examine living or fixed samples, depending on the experimental design and measurement platform. The resulting fluorescence data can also be used to evaluate the apparent effects of experimental treatments on host or pathogen behavior.