Polygon400 Patterned Photostimulators for Optogenetics

* Free demo is ONLY available to US/Canada based universities and government research labs, through an official 'sale or return' order.

Optogenetics is a technology that allows one to investigate behaviours of neurons and their circuits, by precisely delivering light - of different colors and with high spatial and temporal resolutions - to stimulate/silence selected neurons. In addition, there may also be a need to control neural activity independently and/or simultaneously at different locations. For example, ChR2 activates neurons stimulated by blue light (e.g. 470nm), whereas NphR inactivates neurons illuminated by yellow light (e.g. 590nm). By precisely delivering 470nm to specific neurons with millisecond temporal resolution to evoke synaptic transmission between neurons, one can map synaptic circuitry and neurotransmitter receptors. Mightex’s Polygon Digital Patterned Illuminators are specifically designed for such applications.

For more details and/or to place an order
, please click here: Polygon400 Multiwavelength Patterned Illuminators .
   
     This video shows a Polygon400 dynamic spatial illuminator executing a grid pattern sequence stimulation for brain circuit mapping using ChR2 activation.

 

Customer Testimonials and Application Examples -



1. In vivo optogenetic control of zebrafish larva using Polygon 400 illuminator.


A) Light-evoked response of a head-fixed larva expressing channelrhodopsin (right). Photostimulation site was indicated by a blue circle (470 nm).

B) A schematic diagram of the Polygon400 patterned illumination in in-vivo optogenetic mapping system.

(Courtesy Dr. Sachiko Tsuda, Graduate School of Science and Engineering, (Research and Development Bureau) Saitama University, Saitama, Japan.)



2.  Optogenetic activation of channelrhodopsin in transfected hippocampal neurons using Mightex's Polygon400.
Polygon400 for Optogenetics

A) Patch-clamp recording of the current generated by the channelrhodopsin when activated with the blue LED light from the Polygon400 illuminator as indicated by the blue bar.



B) Activation of an action potential (upper trace) and the corresponding current under voltage-clamp conditions (lower trace). The action potential could be evoked by a 0.5ms stimulation of the blue LED light from the Polygon400 illuminator at 60% intensity.

(Courtesy Dr. Hans van Hooft, University of Amsterdam.)



3. Optogenetic stimulation of synaptic inputs using Mightex's Polygon400.
Polygon400 for Optogenetics


Light-evoked inhibitory postsynaptic current (IPSC) recorded in a transgenic mice expressing ChR2 in GABAergic neurons. The postsynaptic cell is non-GABAergic (ChR2 negative) and blue light stimulates GABAergic afferents expressing ChR2. Blue bars indicate the time of light illumination.

Spot 1 illuminated by Polygon400 evoked reliable IPSCs whereas Spot 2 caused no response.

(Courtesy Dr. Wataru Inoue, University of Western Ontario, Canada.)



4. Light-modulated protein-protein interactions on a coverslip surface.



The video shows a protein-protein interaction that is inhibited by blue light exposure. One component is coupled to the coverslip through biotin surface chemistry, the other is labeled with mCherry, and binds to the coverslip in the dark. Patterned illumination with the Polygon 400 results in reversible dissociation of the mCherry-tagged protein, which rebinds within minutes of turning off blue light exposure. The Wittmann lab is working on developing this into a cell adhesion surface that can be controlled by light.

(Courtesy Dr. Torsten Wittmann, University of California San Francisco.)



5. Using Polygon to stimulate hippocampal slices acutely prepared from mice transgenically expressing ChR2 in the dentate gyrus.



The first figure is a repetitive stimulation with a larger circular pattern which elicited increasing responses until I action potential is generated. The second figure is increasing the intensity of stimulation from 30% to 100% and keeping the size pattern the same. Here we're driving the cell at 10 Hz at 100%, and one can see that the stimulation is sufficient to drive doublets of action potentials during each bout of depolarization.


(Courtesy Dr. Geoffrey G. Murphy, Molecular & Behavioral Neuroscience Institute, Department of Molecular & Integrative Physiology, University of Michigan. Recordings made by Dr. Shannon Moore.)



6. Local photostimulation of channelrhodopsin-2 using Polygon400 illuminator.




A) Acute brain slice from a YFP-channelrhodopsin-2 (ChR-2) mouse depicting its expression in cortical L5 pyramidal neurons. B) L5 pyramidal neuron from the somatosensory cortex filled with Alexa 594 to allow visualization of neuronal compartments without stimulating ChR-2. Blue dots indicate on the illuminated areas (Blue LED - 470 nm) along the apical dendrite. Expansion of the marked areas depicting the delicate dendrites that were stimulated. C) Electrophysiological (patch clamp) current recordings from the soma, corresponding to local photostimulation of ChR-2 by blue light. The numbers in the bottom of the trace corresponds to the stimulated dot numbers as indicated in B.

(Courtesy Dr. Yossi Buskila, MARCS Institute, Bioelectronics and Neuroscience, University of Western Sydney, Australia.)



7. Mapping optically induced depolarization in ChR2-expressing hippocampal neurons using Polygon400.



E18 Sprague-Dawley rat neurons were transduced with CamKII-ChR2-GFP lentivirus. Somatic activity was recorded via whole cell patch-clamp electrophysiology. Each field was illuminated by the Polygon400 at 100% power and 20ms exposure time. Intensity of magenta pattern represents depolarization with respect to the instantaneous resting potential prior to stimulation with the Polygon's 470nm LED. In the image, green represents the magnitude of GFP signal and black represents the fluorescence intensity of AlexaFluor 594 backfilled by the patch pipette.


(Courtesy Dr. Jacob T. Robinson, Departments of ECE and BioE, Rice University, Houston, Texas. Data collected and prepared by Dan Murphy, Joel Dapello and Ben Avants.)



8. Photopatterning of an acrylate film.


A 50 μm film of liquid tetra (ethylene glycol) diacrylate, containing 1 wt% Irgacure 819 photoinitiator, was irradiated through a glass coverslip, using the Polygon400 and a 400 nm LED light source to project a pattern onto the film surface. (A) Projection of CU logo image using the 400 nm LED and a 4X objective. (B) Brightfield image of the resulting pattern in the film. Photopolymerization causes a large change in refractive index in the resin, allowing immediate visualization of the pattern. Standard development techniques could subsequently be performed by washing the film in solvent to remove the unexposed areas of liquid resin.

(Courtesy Gayla Berg, Bowman Research Group, Department of Chemical and Biological Engineering, University of Colorado, Boulder, Colorado, USA.)



9. Achieving subcellular resolution of input activation in a 250μm-thick acute cortical section using the Polygon400.



NMDA-receptor mediated synaptic currents recorded at a +40mV membrane potential were elicited by light activation of Channelrhodopsin-expressing terminals of thalamocortical afferents onto an upper layer cortical GABAergic interneuron. Moving the 470nm rectangular illumination to the right or the left of the cell activates inputs of different strength innervating different somatodendritic domains of the cell. In black and red are the individual and the average traces respectively.


(Courtesy Dr. Theofanis Karayannis, Fishell Research Group, Department of Neuroscience & Physiology, New York University, New York, New York, USA.)

 

Polygon400 installation guide
[Note: In general, one may simply skip Step-1, as all Polygon units are pre-aligned to focus at infinity in factory.]

 

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Please call (USA) +1-925-218-1885 or (Canada) +1-416-840-4991,
or email sales@mightexsystems.com or sales@mightex.com.