Fitc filter set

Expression of fluorescent marker genes (i.e., ZsGreen, DsRed, and tdTomato) in plant tissues was observed using a Stemi SVII stereomicroscope equipped with an HBO illuminator (Zeiss, Thornwood, NY, USA), a FITC filter set (λ_excitation = 480 nm, and λ_emission = 515 nm; Chroma Technology, Bellows Falls, VT, USA), and a DsRED filter (λ_excitation = 545/25 nm, dichroic 565LP, λ_emission = 605/70 nm; Chroma Technology). Expression of ZsGreen in isolated embryos was observed under a Zeiss Axioskop 2 plus fluorescence microscope equipped with an 89 North^® PhotoFluor LM-75 illumination system (Chroma Technology) and a FITC filter set. Images were taken using an AxioCam camera (Carl Zeiss, Oberkochen, Germany) and the AxioVision LE64 software.

Live-cell imaging to detect H188 subcellular localization was performed at 23 °C using a Zeiss Axio Observer.Z1 microscope equipped with 100x or 63x Plan-Apochromat Oil DIC objectives (Carl Zeiss, Inc., Thornwood, NY) and interfaced with a Hamamatsu Orca-Flash4.0 LT camera (Hamamatsu Photonics, Bridgewater, NJ), a PhotoFluor LM-75 light source (89 NORTH, Burlington, VT), and an FITC filter set (Chroma Technology Corp.). For co-detection of H188, DNA and F-actin in fixed and permeabilized HEK293-H188-C24 cells, the FITC filter set for detection of H188 was combined with a DAPI filter set for detection of DNA, and a TRITC filter set for detection of rhodamine phalloidin (Sigma-Aldrich, St. Louis, MO) that binds F-actin. MetaMorph imaging software (Molecular Devices) was used to control illumination shutters, camera exposure, and image acquisition. Image stacks were recorded with a Z-distance of 0.1 μm and subjected to deconvolution for construction of major projection overlays.