OPTOGENETIC HARDWARE SETUP

   
  In Vitro Optical Stimulation
     
 

Optical control of cultured neurons and cultured or acute brain slices can be accomplished using a variety of light sources. Below are several examples of light sources that can be used to rapidly switch the wavelength of excitation.
     
  Light sources with shuttering mechanisms  
Sutter Instruments Lambda DG-4 [ Link ]
CAIRN LED Light Source [ Link ]
     
  Shutters for use with existing light sources  
UniBlitz Ultra-fast Shutters [ Link ]
   
  Fluorescence Filters  
Chroma Technology (51022X for ChR2 and NpHR co-excitation) [ Link ]
  Semrock (FF01-475/35-25 for ChR2, FF01-585/40-25 for NpHR) [ Link ]
  Omega Optical [ Link ]
     
  In vivo Optical Stimulation  
     
  Light can be delivered deep into the brain using either lasers or high-intensity LEDs.
 
Optical Fibers: We have developed a method using stereotactic targeting to place optical fibers to precise positions in the rodent brain. The methodology has been reported in Aravanis et al., Adamantidis et al. and Gradinaru et al.
   

To implement the fiber optics setup, a PlasticsOne cannula is surgically implanted on the skull of the test animal using stereotactic guidance. The cannula allows for precise targeting of the optical fiber. To minimize tissue damage, small diameter optical fibers can be ordered through ThorLabs. The optical fiber can then be connected to either a blue or yellow laser via a PC/PC connection. Blue or yellow lasers can be ordered through a number of manufacturers such as Crystal Laser.

The surgical implant is illustrated below, along with a list of parts and vendors:
   
 
   
 

Guide Cannula: C313G/SPC from PlasticsOne (used to guide optical fiber, mount with dental acrylic)
  Internal Cannula: C313I from PlasticsOne (used to virus injection through cannula guide)
  Dummy Cannula: C313DC from Plastics One (used to cover the cannula hole)
 
  Optical Fiber: BFL37-200-CUSTOM from ThorLabs (specify to have length=0.6m, EndA=FC/PC, and EndB=FLAT CLEAVE)
 
  Blue Laser for ChR2 Stimulation: BCL-050-M, 50mW, 473nm, TEMoo from Crystal Laser (specify to have TTL on/off, laser is off when no TTL input, and Fiber couple to multi mode fiber with FC/PC adaptor)
 
  To secure the fiber onto the guide cannula during behavioral experiments, the optical fiber from ThorLabs can be modified to screw onto the guide cannula's thread. See picture below:
 
 
   
  The following movies show optical stimulation of the motor cortex in Thy1::ChR2-EYFP transgenic and wildtype mice. [ Thy-1::ChR2-EYFP Mouse | Wildtype Mouse Control ]