Ex Vivo Optogenetics
Ex Vivo Optogenetics - Related Videos
09:37
In vivo Optogenetic Stimulation of the Rodent Central Nervous System
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Cited by 25 •
2015
Optogenetics has become a powerful tool for use in behavioral neuroscience experiments. This protocol offers a step-by-step guide to the design and set-up of laser systems, and provides a full protocol for carrying out multiple and simultaneous in vivo optogenetic stimulations compatible with most rodent behavioral testing paradigms.
11:13
Ex Vivo Optogenetic Dissection of Fear Circuits in Brain Slices
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Cited by 8 •
2016
Optogenetic approaches are widely used to manipulate neural activity and assess the consequences for brain function. Here, a technique is outlined that upon in vivo expression of the optical activator Channelrhodopsin, allows for ex vivo analysis of synaptic properties of specific long range and local neural connections in fear-related circuits.
06:06
Optogenetic Functional MRI
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Cited by 15 •
2016
This protocol describes the steps and data analysis required to successfully perform optogenetic functional magnetic resonance imaging (ofMRI). ofMRI is a novel technique that combines high-field fMRI readout with optogenetic stimulation, allowing for cell type-specific mapping of functional neural circuits and their dynamics across the whole living brain.
12:00
Selective Viral Transduction of Adult-born Olfactory Neurons for Chronic in vivo Optogenetic Stimulation
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Cited by 7 •
2011
Adult-born neurons of the olfactory bulb can be optogenetically controlled using Channelrhodopsin2-expressing lentiviral injection in the rostral migratory stream and chronic photostimulation with an implanted miniature LED.
13:44
A Method for High Fidelity Optogenetic Control of Individual Pyramidal Neurons In vivo
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Cited by 2 •
2013
The recent development of neuroscience tools that combine genetics and optics, termed "optogenetics", enables control over neural circuit activity with an unprecedented level of spatial and temporal resolution. Here we provide a protocol for integrating in vivo recording with optogenetic manipulation of genetically-defined subsets of prefrontal cortical and subicular pyramidal neurons.
FAQs
The opsin determines how illuminated neurons respond to light. When activated by an appropriate wavelength, these light-sensitive proteins open or close ion channels, changing the neuron’s membrane potential. Researchers can then measure the resulting electrical or synaptic response, allowing optical stimulation to be linked to activity in defined cells within the excised tissue.
Genetic targeting restricts opsin expression to selected neuronal populations rather than stimulating every cell near the light source. This cellular specificity helps investigators associate a response with a defined circuit element. By comparing stimulation with electrical or synaptic recordings, they can examine connectivity and distinguish population-specific contributions to circuit activity.
The ex vivo approach removes the complexity of the intact organism and provides a controlled setting for examining brain tissue directly. Researchers can combine optical stimulation with electrophysiology, imaging, or pharmacology while focusing on cellular and synaptic mechanisms. Findings from this preparation complement in vivo studies by clarifying circuit properties that may contribute to behavior or disease.
Researchers first introduce light-sensitive opsins into selected neurons, then excise the relevant brain tissue and maintain it outside the body. They illuminate the preparation with a chosen wavelength while recording electrical or synaptic responses. The resulting measurements reveal how activating the targeted population influences local circuit function under controlled experimental conditions.
Optical stimulation can be paired with electrophysiology to measure electrical activity or synaptic responses, with imaging to observe associated circuit signals, and with pharmacology to test how chemical manipulation changes those effects. Combining methods gives investigators complementary evidence about connectivity, synaptic function, and the cellular processes responsible for an observed response.
This preparation helps characterize how defined neuronal populations connect, how synapses function, and how circuit activity arises from targeted cellular elements. It is also useful for investigating cellular mechanisms associated with behavior and disease. Because the tissue is studied outside the body, researchers can isolate these mechanisms and test them alongside controlled optical, recording, or pharmacological manipulations.