- White Paper by Annik Yalnizyan-Carson et al. "Patterned Illumination Systems for Optogenetics".
- White Paper: "All-Optical Deep Brain Imaging & Stimulation Tools for In Vivo Neuroscience" has provided an overview of the existing all-optical imaging and stimulation tools, and compared the advantages and disadvantages of these techniques.
Applications in optogenetics
 Researchers at NYU use Mightex's Polygon patterned illuminator and OASIS system to study spatio-temporal codes in the olfactory bulb of awake and behaving mice. For more details, please read "Perceptual Invariance to the Olfactory Spatio-Temporal Code", by Edmund Chong and Dmitry Rinberg.
DMD-based illumination systems
Modifying a DMD-based projector for multiwavelength patterned illumination
Build your own DMD-based photostimulation device
The lab of Rainer Friedrich has published a Nature Protocols paper showing how to build a DMD-based photostimulation device from scratch. The device is particularly well suited for stimulating small isolated preparations such as zebrafish larvae.
- Download the paper here.
The lab of Cha-Min Tang has published a JOVE protocol showing the design and implementation of a DMD-based system and integration on an upright microscope. Optical design and implementation are discussed as well as examples of using the system to uncage glutamate on hippocampal pyramidal neurons.
- Access the JOVE protocol .
Commercially Available Systems
The "Mosaic" system by Andor Technology
The Andor Mosaic 3 system utilizes a DMD to allow for precision photon patterning and sequencing of complex regions of light in the spatial and temporal domains. The system has 4 GB of onboard memory (FPGA) that allows users to upload patterns directly to the DMD head allowing for physiological patterning of light and sequencing.
The system is adaptable to any current generation microscope and each microscope has a patented flat field corrected epi port ensuring very flat field of illumination. The Mosaic 3 can be integrated with most available light sources, including LEDs, traditional Hg lamps, and a variety of lasers can be incorporated.
The Andor Mosaic 3 Duet system allows for simultaneous two color illumination. For instance, Orion Weiner and colleagues have used this system to simultaneous activate PIF-2 (phytochrome) at 650nm in specific regions while at the same time delivering 750nm light to inactivate (Levskaya et al. 2009 - Nature) .
The "Polygon" system by Mightex Systems
Mightex's Polygon400 Patterned Illuminators are designed for advanced bioscience research applications that require precise spatial, temporal, and spectral control of light. The Polygon400 is particularly valuable for the highly-advancing field of optogenetics, as researchers have the need to turn on/off select cells with specific wavelengths of light (e.g. 470nm for ChR2, or 590nm for NphR). Unlike wide-field illumination that illuminates all cells, the Polygon400 allows researchers to precisely target select cellular or sub-cellular features such that researchers will have much better control of neuronal activity. Furthermore, this highly advanced illumination tool can also be used for applications such as FRAP, uncaging and photoactivation etc. The Polygon400's internal memory is capable of storing >1,000 patterns, making it possible for customers to perform sophisticated spatio-temporal sequencing.
Mightex offers three models of the Polygon400 (E-, G-, & DP-Series) that each provide their own unique strengths for specific applications. First, the Polygon400 E-series contains up to three built-in LEDs that range from 400-700nm. This product is the most cost-effective Polygon400 device and is commonly used for optogenetics. Second, the Polygon400 G-Series is compatible with any lightsource that accepts a 3mm core, 5mm ferrule lightguide and can illuminate from 400-700nm. This Polygon400 is useful for applications requiring higher power, such as photoactivation, and optogenetic stimulation across a large area. Lastly, the Polygon400 DP-Series can illuminate light in the UV range (350-700nm) and provides finer spatial resolution illumination. This Polygon400 is useful for applications requiring UV illumination and finer resolution, such as uncaging and photoconversion.
The Polygon400 can be easily mounted on a wide-range of upright and inverted microscopes (e.g. Leica, Nikon, Olympus, Zeiss) using available microscope adapters. Furthermore, the Polygon400 is compatible with Mightex’s OASIS Macro/Micro and OASIS Implant platforms, for performing patterned illumination in awake head-fixed or freely-moving animals, resepectively.
With a USB interface and user-friendly software PolyScan, Polygon400 is extremely easy to install and to operate. Polygon400 is also supported by Nikon's NIS Elements software and MicroManager. Furthermore, the Polygon400 can be easily synchronized with electrophysiological equipment and external TTL devices to synchronize experimental recordings and illumination.
For a list of customer references, please see here.
Here are some examples of the latest publications using the Polygon400:
(1) H. E. Johnson, Y. Goyal et al, “The Spatiotemporal Limits of Developmental Erk Signaling”, Developmental Cell, January 2017;
(2) M. Watanabe et al, “Optogenetic manipulation of anatomical re-entry by light-guided generation of a reversible local conduction block”, Cardiovascular Research (2017) 113, 354–366;
(3) A. Y. Malyshev et al. “Chloride conducting light activated channel GtACR2 can produce both cessation of firing and generation of action potentials in cortical neurons in response to light”, Neuroscience Letters, February 2017;
(4) James L. Butler, Philipe R. F. Mendonc, Hugh P. C. Robinson, and Ole Paulsen, "Intrinsic Cornu Ammonis Area 1 Theta-Nested Gamma Oscillations Induced by Optogenetic Theta Frequency Stimulation", The Journal of Neuroscience, April 13, 2016; and
(5) Natalia V De Marco García, et al "Sensory inputs control the integration of neurogliaform interneurons into cortical circuits", Nature, Feb. 2015.
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