Phase contrast, epifluorescence, and Total Internal Reflection
Fluorescence Microscopy (TIRFM) images were collected on a Nikon Ti-E microscope
using a Nikon CFI Plan Apochromat DM Lambda 100X Oil objective, 1.45 NA, phase
ring Ph3. Cameras used were an ORCA-Flash4.0 V2 sCMOS (Hamamatsu) and an iXon
Ultra 897 EMCCD (Andor). Fluorescence excitation was achieved using a MLC4008
laser launch (Agilent), with a 488 nm laser used for mNeonGreen imaging and a
561 nm laser used for JF549 imaging. For fluorescence emission, a C-NSTORM QUAD
filter cube was used, along with an additional ET525/50m filter for green
emission and ET600/50m filter for red emission (Chroma). The microscope was
enclosed in a chamber heated to 37°C. Images were acquired using
NIS-Elements version 5.02.01.
Fluorescence Microscopy (TIRFM) images were collected on a Nikon Ti-E microscope
using a Nikon CFI Plan Apochromat DM Lambda 100X Oil objective, 1.45 NA, phase
ring Ph3. Cameras used were an ORCA-Flash4.0 V2 sCMOS (Hamamatsu) and an iXon
Ultra 897 EMCCD (Andor). Fluorescence excitation was achieved using a MLC4008
laser launch (Agilent), with a 488 nm laser used for mNeonGreen imaging and a
561 nm laser used for JF549 imaging. For fluorescence emission, a C-NSTORM QUAD
filter cube was used, along with an additional ET525/50m filter for green
emission and ET600/50m filter for red emission (Chroma). The microscope was
enclosed in a chamber heated to 37°C. Images were acquired using
NIS-Elements version 5.02.01.