To specifically activate selected subset of neurons expressing soma-tagged ChR2 in multiple neurons, targeted pattern illumination was performed using a digital micromirror device (Mosaic 2, Andor Technologies UK) mounted on Axio Examiner D1 microscope (Zeiss) connected to an X-LED1 light source.
Nuclear fluorescent tag (mRuby) was used to identify ChR2-expressing neurons and corresponding circular spots (12 µm diameter) were placed on individual cells. To illuminate individual spots with 470 nm blue light, Andor iQ 3.0 software (Andor Technologies, UK) and X-Cite XLED1 software (Lumen Dynamics) were used to control the micromirror array and XLED light source, respectively. IPSCs in response to stimulated spots were recorded at 0 mV in the presence of excitatory blockers (1 µM NBQX, 50 µM L-APV).
Bicuculline methobromide was purchased from Tocris Bioscience (Bristol, UK). All other chemicals and drugs were obtained from Sigma-Aldrich (Missouri, USA).
Nuclear fluorescent tag (mRuby) was used to identify ChR2-expressing neurons and corresponding circular spots (12 µm diameter) were placed on individual cells. To illuminate individual spots with 470 nm blue light, Andor iQ 3.0 software (Andor Technologies, UK) and X-Cite XLED1 software (Lumen Dynamics) were used to control the micromirror array and XLED light source, respectively. IPSCs in response to stimulated spots were recorded at 0 mV in the presence of excitatory blockers (1 µM NBQX, 50 µM L-APV).
Bicuculline methobromide was purchased from Tocris Bioscience (Bristol, UK). All other chemicals and drugs were obtained from Sigma-Aldrich (Missouri, USA).