Optogenetics is an emerging field. This technology can be used to up regulate or down regulate the cellular activity in multiple brain regions of many species and more and more other tissues in vitro or in vivo. It is a very flexible system. Experimental design often requires rich experience, and any incomplete consideration will lead to the failure of the experiment. Creative BioMart provides customized optogenetics experiment design services for customers.
The gene encoding opsin must be under the control of the promoter to stimulate its expression at the required time and place.
For in vitro experiments, we propose to achieve it by transfection (by electroporation, calcium phosphate or liposome).
For in vivo work, constructs are usually packaged as viruses and then injected into the target region (virus transduction). Alternatively, transgenic mice expressing opsin under the control of a specific promoter can be produced.
For in vivo experiments, this is usually coupling an LED or laser to a fiber optic cable to transmit light to a precise area.
The selected methods may include electrophysiological recording, calcium imaging, optical functional magnetic resonance imaging, molecular biology or behavioral testing, depending on the specific goal of the experiment.
When designing photogenetics experiments, opsin and delivery system need to be selected at the same time. The factors we will consider include but are not limited to
Please tell us whether your goal is to turn the switch on or off in the experiment. We will choose excitatory or inhibitory opsin according to your needs.
From purple to red, there are many different activation wavelengths that can be used in optogenetic systems. However, each wavelength of light has different characteristics. For example, if you plan to place an optical fiber in the brain of an experimental animal, we will recommend that you choose red light, because red light shows better tissue permeability, thus reducing the invasiveness of the experimental process. Different activation wavelengths also make it possible to combine multiple opsins in the same experiment.
Time accuracy is the key of photogenetics experiment. We design the time of light stimulation according to whether you need short-term or long-term neuronal activation / inactivation. These can range from milliseconds to "long-lasting".
Opsin expression and illumination regions determine the population of neurons to be manipulated in a given experiment. Generally, the most robust and stable expression is achieved in transgenic mouse lines. Opsin delivery using viral vectors leads to more local expression of opsin. Depending on the virus and promoter system used, an incubation time is required before reaching the peak expression of opsin.
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