Zen blue image processing software

Ten beetles of each sex, collected in traps during field bioassays in 2014, were examined using an Olympus SZX7 stereomicroscope to assess presence/absence of pores in males and females. Images were taken using a Canon EOS 70D camera attached to a light microscope digital SLR adapter on a Nikon SMZ1000 stereomicroscope equipped with a Plan Apo 1X WD70 objective lens (8–80x total magnification).
Specimens for histological sectioning were collected in live traps (described below), placed adjacent to the Adams Produce site (see below for details about field site). Specimens were submerged in ethanol-based hand sanitizer as a fixative. Fixation, sectioning, and imaging were contracted to Laudier Histology (New York, NY). Images were acquired from microscope slides with a Carl Zeiss AxioImager Z1 microscope, running ZEN Blue image processing software (Zeiss, Oberkochen, Germany).

Murine hair follicles for IF assays were obtained from HsdWin:NMRI mice (Envigo, UK). All samples were fixed in 10% neutral buffered formalin (Sigma-Aldrich) for 20 min and permeabilized with 0.5%TritonX-100 (Carl Roth GmbH + Co. KG, Karlsruhe, Germany) in phosphate-buffered saline (PBS) for 10 min. After every step, samples were washed in PBS (2 x 2 min). Then they were blocked in PBS containing 1% bovine serum albumin (Sigma-Aldrich) for 20 min. The blocking solution was also used to dilute the antibodies. Samples were incubated in primary antibody solution overnight at 4° C. After washing with PBS, samples were incubated in secondary antibody solution for one hour. The primary and secondary antibodies are listed in Supplementary Tables 2, 3. Samples were washed in PBS and mounted using Vectashield antifade medium with DAPI (Vector Laboratories, Burlingame, CA, USA). Images of mouse hair follicles were obtained using an Axio Observer.Z1/Cell Observer Spinning Disc microscopic system (Zeiss, Oberkochen, Germany) with a 63x oil objective. All images were processed using ZEN Blue Image processing software (Zeiss).