Channelrhodopsin2-yfp
Channelrhodopsin2-yfp - Related Videos
08:39
Whole-cell Patch-clamp Recordings for Electrophysiological Determination of Ion Selectivity in Channelrhodopsins
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Cited by 26 •
2017
This article describes how the ion selectivity of channelrhodopsin is determined with electrophysiological whole-cell patch-clamp recordings using HEK293 cells. Here, the experimental procedure for investigating chloride selectivity of an anion-selective channelrhodopsin is demonstrated. However, the procedure is transferable to other channelrhodopsins of distinct selectivity.
10:03
Multiphoton Microscopy of Cleared Mouse Brain Expressing YFP
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Cited by 11 •
2012
Multiphoton microscopy of whole mouse organs is possible by optically clearing the organ before imaging, but not all protocols preserve the fluorescent signal of fluorescent proteins. Using an optical clearing method with ethanol-based dehydration and benzyl alcohol:benzyl benzoate clearing, we show high-resolution multiphoton images of whole mouse brain expressing YFP.
07:04
Long-range Channelrhodopsin-assisted Circuit Mapping of Inferior Colliculus Neurons with Blue and Red-shifted Channelrhodopsins
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Cited by 4 •
2020
Channelrhodopsin-assisted circuit mapping (CRACM) is a precision technique for functional mapping of long-range neuronal projections between anatomically and/or genetically identified groups of neurons. Here, we describe how to utilize CRACM to map auditory brainstem connections, including the use of a red-shifted opsin, ChrimsonR.
08:31
FM Dye Cycling at the Synapse: Comparing High Potassium Depolarization, Electrical and Channelrhodopsin Stimulation
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Cited by 7 •
2018
Synaptic vesicle (SV) cycling is the core mechanism of intercellular communication at neuronal synapses. FM dye uptake and release are the primary means of quantitatively assaying SV endo- and exocytosis. Here, we compare all the stimulation methods to drive FM1-43 cycling at the Drosophila neuromuscular junction (NMJ) model synapse.
01:13
meta-Directing Deactivators: –NO2, –CN, –CHO, –CO2R, –COR, –CO2H
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2023
All meta-directing substituents are deactivating groups. These substituents withdraw electrons from the aromatic ring, making the ring less reactive toward electrophilic substitution. For example, the nitration of nitrobenzene is 100,000 times slower than that of benzene because of the deactivating effect of the nitro group. The first step in an electrophilic aromatic substitution is the addition of an electrophile to form a resonance-stabilized carbocation. The energy diagrams for the...
FAQs
Blue-light activation changes the membrane's electrical state through ion-channel gating. When Channelrhodopsin-2 opens, positively charged ions cross the membrane, producing depolarization in neurons that express the fusion protein. This shift can influence neuronal electrical activity, giving investigators a direct way to examine how stimulating a defined population alters downstream neural function.
YFP supplies a fluorescence readout rather than the light-gated activity itself. By visualizing YFP, investigators can identify cells expressing Channelrhodopsin2-YFP and determine where the tagged protein is located with fluorescence microscopy. This distinction helps connect an optical manipulation to the cells actually targeted, strengthening interpretation of neural activity experiments.
Optical stimulation turns the construct into an intervention, while YFP identifies the cells receiving that intervention. Researchers can therefore relate a controlled change in activity to a measured consequence, rather than only correlating naturally occurring neural signals with behavior or circuit state. This design is especially useful when testing whether a defined neuronal population contributes to a function.
A basic workflow begins by locating YFP-positive cells through fluorescence microscopy. Researchers then use blue light to activate Channelrhodopsin-2 in the identified population and examine how neuronal electrical activity or a relevant neural outcome changes. The linked visualization and stimulation steps help confirm that the experimental manipulation is directed at cells expressing the intended construct.
A response is most informative when considered alongside evidence that the stimulated cells express the construct. YFP fluorescence provides that expression and localization context, while blue-light exposure tests whether activating those cells changes neuronal electrical activity or a related neural function. Together, these observations connect targeting, stimulation, and outcome in one experiment.
This construct is suited to experiments asking how defined neuronal activity affects neural circuits, behavior, sensory processing, or disease-related network function. Its value lies in combining cell identification with optical control, so investigators can manipulate a selected expressing population and evaluate the resulting change in a specific biological context.
Fluorescent labeling can show where expressing cells are, but it does not by itself provide the optical activation described here. Channelrhodopsin2-YFP adds a light-responsive route for changing activity in those identified neurons. That combination supports experiments that move from locating a population to testing its contribution to circuit or behavioral outcomes.