Dmc2900 camera

The holotype of Conobregmabradpitti sp. n. is deposited in the Hymenoptera Institute Collection, Department of Entomology, University of Kentucky, Lexington, Kentucky. It was imaged using an Olympus SXZ16 microscope with automated multiple image capture at preset focal levels using an Olympus DP72 camera, and image combination using the Cell^D image processing system. The specimen was card-mounted and rather fragile but we successfully remounted it to enable more features to be seen.
The holotype of Facitorusnasseri sp. n. is deposited in the Department of Zoology, University of Calicut, Kerala, India. It was imaged using an Leica M205A stereomicroPageBreakscope with automated multiple image capture at preset focal levels using an Leica DMC 2900 camera, and image combination using the Leica Application Suite image processing system v4.7. All images were edited using Photoshop CS6 (Version 6.1) (Adobe Inc.).
Terminology follows van Achterberg (1988) except for wing venation nomenclature which follows Sharkey and Wharton (1997) ; see also Figure 2.2 in Quicke (2015) for comparison of wing venation naming systems.

Giemsa-stained culture smears were imaged on a DM2000 LED light microscope (Leica) equipped with a DMC2900 camera (Leica). Fluorescence microscopy was performed with an Eclipse Ni light microscope (Nikon) fitted with a C11440 camera (Hamamatsu) or with a D6B fluorescence microscope (Leica) equipped with a DFC9000 GT camera (Leica). For detailed microscopy protocols including quantitative analysis of Fluo-CQ uptake, please refer to SI Appendix.

Full necropsies were performed on all fetuses. Representative peripheral tissues, included in Supplemental Table S1, were collected and fixed in 10% neutral buffered formalin. Before removal from the cranium, the brain was flushed with normal saline followed by 4% paraformaldehyde (PFA), and then immersion-fixed in 4% PFA. Tissues were processed routinely, embedded in paraffin, sectioned at 4 µm (peripheral) or 5 µm (brain), and stained with hematoxylin and eosin for analysis by light microscopy. All peripheral fetal tissues were examined by ONPRC veterinary pathologists. Photomicroscopy was performed with a Leica DMC2900 camera and Leica LAS X microscopic imaging software. Slides were scanned using a Leica Aperio AT2 slide scanner. Histologic sections of the brain underwent routine histological examination by two neuropathologists (M.H.H. and M.R.G) blinded to exposure and outcomes.

We obtained paraffin blocks of all prostatic lobes containing WT and tumoral samples from GEMM from David Neal’s Uro-Oncology Group at CRUK Cambridge Institute (University of Cambridge, UK). At least 10 different urogenital complex paraffin blocks from the wild type and 20 from both knockout mice were sectioned. Histological sections of the prostate at various stages of development and progression were deparaffinized, hydrated, and washed in PBS (0.1 M, pH 7.4). Antigen retrieval was performed using 10 mM citrate buffer pH 6.0 for 35 min in a Dako Cytomatica pressure cooker. Subsequently, slices were submitted to endogenous peroxidase blockade with 3% H₂O₂ solution in methanol for 10 min, protein–protein interaction block with 3% BSA in PBS, and overnight incubation at 4 °C with primary antibodies against SDC-1 (AB128936), -2 (AB79978), -3 (AB191308), -4 (AB24511), or SDCBP (AB-19903), diluted 1:100 in 1% BSA solution in PBS. The five antibodies were purchased from Abcam (Cambridge, UK). After washing with PBS, the sections were exposed to the peroxidase-conjugated secondary antibody, developed using diaminobenzidine as a chromogen, and counterstained with hematoxylin. We analyzed slides using a Leica DM2500 microscope and acquired images with a Leica DMC2900 camera and Leica Qwin image analysis software version 3.1.

For confocal imaging, live zebrafish embryos were mounted in 0.02% tricaine and 1% low melt agarose, and imaged with a Zeiss LSM 710 or 800 confocal microscope. For in situ hybridization images, fixed and stained embryos were placed in 100% glycerol and imaged with a Leica M165 FC Stereomicroscope, with attached Leica DMC2900 camera.