The LYUOR-3480-96RB in vitro optogenetics light source has 96 built-in 470nm LEDs, which are suitable for 96-well plate irradiation, and can provide irradiation power with an irradiation intensity of 0-9MW, and the light intensity is adjusted by a knob to meet the in vitro optogenetics experiments of CHR2 photosensitive protein.
The LYUOR-3480-150RB in vitro optogenetics light source has 256 built-in 470nm LEDs, which are suitable for irradiation of 150mm Petri dishes, and can provide irradiation power with an irradiation intensity of 0-9MW, and the light intensity is adjusted by a knob to meet the in vitro optogenetics experiments of CHR2 photosensitive protein.
The 470nm narrow band LED meets CHR2 irradiation requirements.
0-9MW irradiation power adjustable.
Irradiation for a long time, the temperature is controlled below 37 degrees Celsius.
A choice of 96-well plates and 15 cm plates can be illuminated as light sources.
The irradiation light source and controller line are 2 meters long, which can meet the irradiation requirements of placing the irradiation light source in the cell culture incubator.
The waterproof design of the irradiation light source can meet the high humidity environment irradiation of the incubator.
256 LEDs are irradiated, and the light intensity in the irradiated surface is uniform.
Common photosensitizing proteins:
1. Activated photosensitive protein:
1) CHR2 (H134R): A mutant of Channelrhodopsin-2 (CHR2), which belongs to the photocontrolled cation channel, which opens the cation channel under the excitation of blue light (preferably at the activation wavelength of 450nm), causing the influx of extracellular cations. Compared with CHR2, CHR2 (H134R) has a higher light sensitivity and a slower channel closure speed, thus increasing the photoelectric signal, but this also makes it less accurate in time than CHR2.
2) CHTA: Another mutant of CHR2, CHR2 (E123T), is a cation channel protein. The channel dynamics of CHETA are faster than those of CHR2, reducing the amplitude of photocurrent. However, the light sensitivity is greatly reduced, it is suitable for high-frequency light stimulation, and the absorption spectrum is red-shifted (preferably at 490 nm) and can be synchronized with calcium ion imaging experiments.
3) C1V1(T T): It is a chimeric mutant of CHR1 from Chlamydomonas and VCHR1 from Chlamydomonas, containing two amino acid mutations—E122T and E162T, so it is called C1V1(T T). It is a cation channel protein driven by yellow light (preferably at a wavelength of 535 nm).
2. Inhibitory photosensitive protein:
1） enphr3.0: ** for the halophilic bacterium natronomonas halorhodopsin (nphr) isolated from saline-alkali archaea, which is a chloride ion pump and belongs to the inhibitory photosensitive protein. NPHR was overexpressed in mammalian cells and localized to the endoplasmic reticulum, and the upper endoplasmic reticulum output element and KIR2 were added after modification1. The upper membrane element of the potassium ion channel, so that it can be anchored to the neuronal cell membrane, and enhance the photocurrent, upgraded to Enphr30。Stimulation of yellow light (preferably at a wavelength of 590 nm) opens ion channels, causing Cl- influx, resulting in an intracellular hyperpolarization response, which in turn inhibits nerve cell activity. enphr3.0 is one of the most widely used inhibitory photosensitizing proteins.
2）arch3.0: An upgraded version of archaerhodopsin (archaerhodopsin, arch) produced by Rhododobacterium salinus, a proton pump that mediates intracellular H+ efflux in response to yellow or green light (preferably at a wavelength of 566 nm). Compared with equivalent versions of other inhibitory photosensitizing proteins, ARCH has higher photocurrent and light sensitivity, and can be expressed in large quantities in the axonal plasma membrane. However, by adding endoplasmic reticulum output elements and neuronal axon-targeting sequences, its functionality was further enhanced and upgraded to ARC30, which can be applied to the fast suppression of axon signal output. This also makes arch30 enables robust, rapid, and reversible synaptic inhibition.