Orca ag ccd camera

CC2-DMPE and DiBAC4(3) ratiometric voltage reporter dyes were obtained from Invitrogen and used as per the standard protocol (Adams and Levin, 2013 (link)). Briefly, CC2-DMPE stock (5 mM) was dissolved 1:1,000 in 0.1× MMR and the embryos were incubated in the dark in this solution for at least 1 h followed by five washes with 0.1× MMR. DiBAC₄(3) stock (1.9 mM) was dissolved 1:1,000 in 0.1× MMR and the CC2-DMPE-stained embryos were then incubated in the dark in this solution for at least 30 min washed thoroughly in 0.1× MMR followed by visualization under the microscope. An Olympus BX-61 microscope equipped with a Hamamatsu ORCA AG CCD camera and controlled by MetaMorph software (Molecular Devices), was used to collect images. ImageJ was used to quantify the fluorescence intensities of the CC2-DMPE: DiBAC signal along the red line across the image as indicated in the illustrations in Figures 4G, 9E). Fluorescence values at each point along this line were used to plot graphs.

CC2-DMPE and DiBAC₄(3) voltage reporter dyes were obtained from Invitrogen and used as per the standard protocol, including dark-field and flat-field correction (Adams and Levin, 2012 (link)). Briefly, the use of two dyes with opposite emission profiles simultaneously provides an internal control and allows ratiometric normalization. CC2-DMPE stock (5 mM) was dissolved 1:1000 in 0.1× MMR and the embryos were incubated in dark in this solution for at least 1 h followed by washes with 0.1× MMR. DiBAC₄(3) stock (1.9 mM) was dissolved 1:4000 in 0.1× MMR and the CC2-DMPE-stained embryos were then incubated in dark in this solution for at least 30 min followed by visualization under the microscope. An Olympus BX-61 microscope equipped with a Hamamatsu ORCA AG CCD camera, and controlled by Metamorph software (Molecular Devices), was used to collect signal. NIH Image J software was used to quantify the fluorescence intensities of the CC2-DMPE:DiBAC signal.

CC2-DMPE and DiBAC₄(3) voltage reporter dyes were obtained from Invitrogen and used as per the standard protocol, including dark-field and flat-field correction^{63 (link)}. Briefly, the use of two dyes with opposite emission profiles simultaneously provides an internal control and allows ratiometric normalization. CC2-DMPE stock (5 mM) was dissolved 1:1000 in 0.1 × MMR and the embryos were incubated in the dark in this solution for at least 1 h followed by 5 washes with 0.1 × MMR. DiBAC₄(3) stock (1.9 mM) was dissolved 1:1000 in 0.1 × MMR and the CC2-DMPE-stained embryos were then incubated in the dark in this solution for at least 30 min washed thoroughly in 0.1 × MMR, followed by visualization under the microscope. An Olympus BX-61 microscope equipped with a Hamamatsu ORCA AG CCD camera, and controlled by MetaMorph software (Molecular Devices), was used to collect signal. NIH ImageJ software was used to quantify the fluorescence intensities of the CC2-DMPE:DiBAC₄(3) signal.