Zen 2 blue software

Live imaging of embryos and larvae was performed using a 60× 1.42NA Plan‐Apochromat objective on a DeltaVision 2 Ultra microscope, equipped with 7‐Color SSI module and sCMOS camera and controlled by Acquire Ultra acquisition software (GE Healthcare). Worms were anesthetized using 10 mM tetramisole in M9 for 5 min before being mounted on 2% agarose pads and imaged at room temperature. Embryos were mounted directly on agarose pads. 0.2 μm GFP/mCherry z‐series as well as single plane DIC images were collected for the amphid and phasmid neurons closest to the objective. For each condition and worm stage, at least 10 worms/embryos were examined. Image stacks were deconvolved using Softworx and then imported into Fiji for postacquisition processing. For strains expressing OSM‐6:GFP, 0.5 μm z‐series were acquired on a Zeiss Axio Imager Z2 microscope (described above) using a Photometrics CoolSNAP‐HQ2 cooled CCD camera controlled by ZEN 2 blue software (Zeiss).
Quantitation of phasmid dendrite lengths (from cell body to ciliary base) and ciliary length (from base to tip) was performed in Fiji using the Segmented Line tool. For each condition, a minimum of 20 worms were examined.

Images and image stacks of sections were acquired using an Axio Imager. Z1 equipped with an ApoTome.2 slider for optical sectioning (Zeiss, Germany). For image processing the ZEN2 blue software (Zeiss, Germany) was used and either single images or extended depth of focus projection of z-stacks were displayed. Whole mount larvae were either imaged as described (α-NeuN staining) or with a light sheet microscope (α-Cldn5 staining, morpholino experiments). For light sheet microscopy, a Lightsheet Z1 (Zeiss) enabled for dual side illumination and equipped with a 20x detection objective (W Plan-Apochromat, numerical aperture = 1.0) and a sCMOS pco. edge 4.2 camera was employed. Image processing consisting of dual side fusion, brightness/contrast adjustment and unsharp masking was performed by using the Zen 3.1 (blue edition) software (Zeiss, Germany). For three-dimensional reconstruction the 3Dxl rendering module (powered by arivis, Germany) was used.

24-hour cultures of C. albicans strains in YPD medium (28°C; 120 rpm) were centrifuged (5 min, 2260 x g). The collected cells were washed with fresh YPD medium and subsequently resuspended in YPD medium to OD₆₀₀ = 0.4. The exposure to AMF was performed in 8-well culture chambers, as described in “Exposure of C. albicans to AMF”. The hyphal transition was induced by cells suspension treatment with FBS (final conc. = 10%) for 2h at 28°C. After incubation, the samples were observed under a Zeiss Axio Imager A2 microscope equipped with a Zeiss Axiocam 503 mono microscope camera (Zeiss, Oberkochen, Germany) for the assessment of cell morphology (n = 50–100 cells in three repetitions). Zeiss ZEN 2 Blue software was used for the measurement of the length (μm) of straight hyphae.

Differential interference contrast (DIC) and fluorescence images were taken by an inverted confocal microscope (LSM880; Carl Zeiss) using a 100× 1.46-NA oil objective lens. Imaging was performed on animals of the same developmental stage and on a similar body region of each animal, unless otherwise indicated. Images were processed and analyzed with ZEN 2 blue software (Carl Zeiss) or Image J (National Institutes of Health). All images were taken at 20 °C.