Applicationsuite x software las x

The samples were analyzed by laser scanning confocal microscopy (Olympus,
FluoView 500) using argon laser 488 nm excitation. Fluorescence emission
of GFP was recorded at 500–520 nm. For 3D images, acquisition
used a Leica SP8 laser scanning microscope (Leica, Wetzlar, Germany)
equipped with a solid state laser with 488 nm light, HC PL APO CS
20×/0.75 objective (Leica, Wetzlar, Germany) and Leica Application
Suite X software (LAS X, Leica, Wetzlar, Germany). Imaging of the
GFP signal was done using the argon laser, and the emission was detected
in a range of 500–525 nm. Autofluorescence of the chloroplasts
was detected in a range of 650–700 nm. For Metarhizium conidia counting, image stacks were first projected using a Z projection
(as maximum intensity) to find all the fluorescent conidia, then counted
by a cell counter using Fiji software.^{54 (link)} The droplet average diameter was measured for every sample by the
particles analysis tool of Fiji software based on confocal microscopy
images. 12 droplets were sampled from each image and plotted as a
3D graph with Origin (OriginLab, Northampton, MA).