For emission ratio imaging, the Dual-View Micro-Imager (Optical Insights) was used.
To avoid bleaching, a Multispec MicroimagerDualView beam splitter (Optical Insights, Tucson, AZ) was installed for simultaneous detection of two emission wavelengths on the CCD chip.
The Dual-View system (Optical Insights, Santa Fe, NM) was used for the simultaneous acquisition of two fluorescence images (donor and FRET).
Cyan and yellow fluorescence were simultaneously collected in a single image using an Optical Insights Dual-view system (Photometrics Headquarters, Tucson, AZ) with a CFP/YFP beamsplitter.
Two emission wavelengths were collected simultaneously by using a Dual-View image splitter (Optical Insights) with appropriate yellow (535 nm/30) and cyan emission filters (480 nm/480.
A Leica inverted microscope was integrated with a magnetic twisting device and a Dual-View system (Optical Insights, Tucson, AZ) to simultaneously acquire both CFP and YFP emission images in response to stress.
Cell fluorescence was split into YFP and CFP channels using a DualView (Optical Insights) equipped with 535/40 and 480/30 emission filters and monitored by the ORCA-ER CCD camera (Hamamatsu, Welwyn Garden City, UK).
All mirrors and filters were obtained from Chroma Technologies (Brattleboro, VT). Green and red emissions were collected simultaneously using a dual-view splitter (Optical Insights, Santa Fe, NM) equipped with a 515/30-nm band-pass filter to collect green emission, a 550-nm dichroic mirror to split the emission, and a 580-nm long-pass filter to collect red emission.
YFP and CFP images were acquired simultaneously using a Dual View image splitter (Optical Insights, Tucson, AZ) equipped with a 505 nm long-pass dichroic filter to separate the CFP and YFP signals, a CFP emission filter (480/30) and a YFP emission filter (535/40).
Simultaneous acquisition of the two colors is made with the DUAL-view beam splitter (Optical Insights, LLC, Tucson, AZ).
