Ph-sensitive Gfp Marker
Ph-sensitive Gfp Marker - Related Videos
06:19
In Vivo Imaging of Cx3cr1gfp/gfp Reporter Mice with Spectral-domain Optical Coherence Tomography and Scanning Laser Ophthalmoscopy
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Cited by 5 •
2017
This protocol describes how high-resolution imaging techniques such as spectral domain optical coherence tomography and scanning laser ophthalmoscopy can be utilized in small rodents, using an ophthalmic imaging platform system, to obtain information on retinal thickness and microglial cell distribution, respectively.
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.
13:47
TIRFM and pH-sensitive GFP-probes to Evaluate Neurotransmitter Vesicle Dynamics in SH-SY5Y Neuroblastoma Cells: Cell Imaging and Data Analysis
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Cited by 8 •
2015
This paper provides a method for investigating neurotransmitter vesicle dynamics in neuroblastoma cells, using a synaptobrevin2-pHluorin construct and Total Internal Reflection Fluorescence Microscopy. The strategy developed for image processing and data analysis is also reported.
09:16
High-sensitivity Detection of Micrometastases Generated by GFP Lentivirus-transduced Organoids Cultured from a Patient-derived Colon Tumor
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Cited by 10 •
2018
To allow highly sensitive detection of the disseminating human colorectal cancer (CRC) cells colonizing tissues, we herein show a protocol for efficient transduction of green fluorescent protein (GFP) lentiviral particles into PDX-derived CRC organoid cells prior to their injection into recipient mice, with stereo-fluorescence microscopic observation.
07:30
Electrophysiological Characterization of GFP-Expressing Cell Populations in the Intact Retina
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Cited by 5 •
2011
This article depicts the recording of individual cells from fluorescently tagged neuronal populations in the intact mouse retina. By using two-photon infrared excitation transgenetically labeled cells were targeted for patch-clamp recording to study their light responses, receptive field properties, and morphology.
FAQs
The GFP chromophore, the light-absorbing part of the reporter, becomes protonated in the acidic synaptic-vesicle lumen. This chemical state suppresses fluorescence while the probe remains inside the vesicle. When exocytosis exposes the reporter to a near-neutral environment, deprotonation restores brightness, converting a local pH change into an optical signal that can be measured in living neurons.
Exocytosis temporarily connects a synaptic vesicle with the extracellular space, replacing its acidic lumenal environment with near-neutral medium. The attached reporter therefore changes brightness quickly rather than requiring a slower downstream cellular response. This rapid transition allows fluorescence changes to mark the timing of vesicle fusion and helps resolve presynaptic events with high temporal resolution.
A sudden fluorescence increase indicates that a previously quenched reporter has encountered near-neutral conditions during vesicle fusion. Subsequent changes can be followed as the vesicle is retrieved, trafficked, and recycled, although the optical pattern must be interpreted in relation to these stages. This makes the marker useful for examining both the initial release event and later presynaptic membrane handling.
The reporter signal depends primarily on the acidity surrounding its chromophore. Acidic conditions suppress fluorescence, whereas near-neutral conditions increase it, so changes in vesicle lumenal pH or exposure to extracellular medium can alter brightness. Neural activity is relevant because it can drive vesicle fusion and thereby change the reporter’s chemical environment, producing measurable fluorescence dynamics.
Researchers genetically express the fluorescent reporter in living neurons, observe fluorescence at presynaptic sites, and monitor brightness changes over time. Optical recordings can then be related to neural activity to identify vesicle fusion and follow retrieval or recycling. The resulting time-dependent signal provides a way to examine presynaptic behavior without relying only on fixed-cell observations.
Fluorescence recordings can reveal when synaptic vesicles fuse, how signals change during retrieval and trafficking, and how efficiently vesicles participate in recycling. Because the measurements occur in living cells with high temporal resolution, they can connect presynaptic membrane events to neuronal communication and activity-dependent changes. This makes the approach valuable for studying dynamic synaptic function.
These reporters link a biochemical property of synaptic vesicles to optical measurements in living neurons. Their signals can expose changes in presynaptic communication and vesicle recycling that may accompany altered neural activity or disease-associated dysfunction. By tracking dynamic events rather than only endpoint structures, researchers can investigate how neuronal signaling changes over time.