Authors
Dario Campagner, Ruben Vale, Yu Lin Tan, Panagiota Iordanidou, Oriol Pavón Arocas, Federico Claudi, A. Vanessa Stempel, Sepiedeh Keshavarzi, Rasmus S. Petersen, Troy W. Margrie, Tiago Branco
Topic
Neuroscience
Extract
"For CRACM experiments, expression of ChR2-mCherrywas assessed based on fluorescence from mCherry expression using LED illumination (pE-100, CoolLED) at a wavelength of 565 nm. Target cells were identified based on fluorescence from EYFP expression using LED illumination (pE-100, CoolLED) at a wavelength of 490 nm. ChR2 was activated with wide-field 490-nm LED illumination (pe-100, CoolLED) using a train of five 1-ms long pulses at 20 Hz (maximum light intensity = 6 mW). The resting membrane potential was determined immediately after establishing the whole-cell configuration and experiments were continued only if cells had a resting membrane potential more hyperpolarized than −45 mV. For dual opsin-assisted circuit mapping and opto-tagging experiments, target putative vGluT2+ cells were identified based on the fluorescence from Gcamp7f expression using 470nm wide-field LED illumination (pE-800, CoolLED) filtered by a 466/40nm bandpass filter (FF01-466/40-25, Semrock). ChR2 was activated with wide-field 435-nm LED illumination (pe-800, CoolLED) filtered by a 430/40nm bandpass filter (86349, Edmund Optics) using a train of five 1-ms long pulses (maximum light intensity = 13.5 mW), that was preceded by a 250ms long pulse of 635-nm wide-field LED illumination (pe-800, CoolLED)50 filtered by a 624/40nm bandpass filter (67035, Edmund Optics), maximum light intensity = 13 mW,. ChrimsonR was activated with wide-field 635-nm LED illumination (pe-800, CoolLED) filtered by a 624/40nm bandpass filter (67035, Edmund Optics) using a train of five 1-ms long pulses (maximum light intensity = 13 mW)."
Product Type
Journal
Nature
Year of Publication
2023
