Dear all, The Ch1 from the Zeiss Exciter is not working anymore. This influences mainly the 2-colour imaging , and mainly your red dye or fluorophore. One colour imaging can be done by sending everything to the first detector (Ch2), meaning that reuse of the settings of a previous image could need some adaptation. Two colour imaging could be done by FRAME sequential imaging : both your dyes will be captured at the same channel, but one after the other. Keep in mind that there is more time-delay between imaging your 2 colours compared to the simultanous or line sequential mode, which is normally used. During live mode I would recommand to only use one channel at a time as it takes more time to see the effect in the second channel. This can be easily done in the software by activating-deactivating a channel (also explained in the ppt in attach). More information in attach and also in the help folder, including a picture with settings allowing to reload the settings for GFP - PI FRAME sequential imaging: S:\research\images\images_cameras\confocal\Help Zeiss Exciter\20191108_Frame sequential scanning GFP PI\ GFP and PI_Frame sequential.lsm Don't hesitate to contact Daniel or me if you would have problems with the lightpath setup. I will keep you update about the repair of this channel. Gr, Evelien Evelien Mylle - Labmanager/ microscopy technician Advanced Live Cell Imaging VIB-UGent Center for Plant Systems Biology Ghent University Technologiepark-Zwijnaarde 71 - 9052 Ghent - Belgium Tel. +32(0)9 331 39 74 [ http://www.psb.ugent.be/ | www.psb.ugent.be ] Genome editing, cutting-edge technology for a sustainable agriculture