With Sub-Cellular Resolution
Photostimulation:
Cell Biology Optogenetics
Overview

Optogenetic toolkits have expanded to offer cell biologists unprecedented spatiotemporal molecular and cellular control. From cell migration to gene regulation or developmental processes, cell biologists now have the ability to elicit specific biological responses with an incredible amount of precision in specific cells within organisms or subcellular regions of individual cells using optogenetics. Specifically, researchers are able to use the methodology depicted below to investigate multiple spatiotemporal biological responses.

single-colour optogenetics cell biology
dual-colour optogenetics cell biology

Key Requirements

sub-cellular resolution to stimulate small regions within cells
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synchronization with imaging equipment and cameras

System Configurations

(Click below to view the relevant system configuration)

(1) Single-Colour Optogenetics

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This configuration based on Mightex’s Polygon400 provides:

1) Single-colour (e.g.CRY2) sub-cell resolution optogenetics of any optogenetic probe.

2) Synchronization with imaging equipment and camera.

3) Integrates into any commercial microscope (inverted or upright).

Components: (1) Polygon400, (2) Microscope Adapter, (3) Any Light Source, (4) Light Source Controller(s), & (5) Any Third-Party Camera.

(2) Dual-Colour Optogenetics

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This configuration based on Mightex’s Polygon400 provides:

1) Dual-colour (e.g. PHYB/PIF, Dronpa) sub-cell resolution optogenetics of any optogenetic probe.

2) Stimulate independent regions with different colours.

3) Synchronization with imaging equipment and camera.

4) Integrates into any commercial microscope  (inverted or upright).

Components: (1) Polygon400 x2, (2) Microscope Adapter, (3) Any Light Source, (4) Light Source Controller(s), & (5) Any Third-Party Camera.

Main Component

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Additional Components

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Examples

Cellular-Resolution Optogenetics

This is a video of an NIH-3T3 expressing a novel optogenetic system termed PixELLs. In the dark PixELLs undergo protein phase separation forming liquid-like clusters as can be seen in the beginning of the movie. However, upon 450 nm light stimulation PixELLs dissolve and become diffuse. We used this PixELL system and the unique capabilities of Mightex’s Polygon400 to test an outstanding theory in the field of protein phase separation. We drew an ROI on the cell to stimulate with a gradient of blue light intensities from Mightex’s Polygon400 for 30 min. This type of control over the intensity and spatial range of illumination would not be possible using other forms of illumination available for confocal microscopy. This work was published in Cell Systems (Courtesy of Elliot Dine and Dr. Jared Toettcher from Princeton University).

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