Mz16 stereomicroscope

Photographs of the ants including dorsal, lateral and full-face views of workers and queens were taken at the MCZC with an imaging system that consisted of a Leica MZ16 stereo microscope, a Leica DCF 420 digital camera, and the Auto-Montage Professional software Leica Application Suite 3.7 and Helicon Focus 5.1; and at the USNM with an imaging system that consisted of a Leica Z16APO microscope and a JVC KY-F75U digital camera with a Leica Motor-focus System attached to an IBM IntelM Pro computer, on which composite images were assembled using Auto-Montage Pro Version 5.03.0018 BETA (Synoptics Ltd.). Scanning electron micrographs were taken with a LaB6 electron source. Images were processed with Adobe Photoshop CS. The distribution map was created using the software ArcGIS v10.1 (Esri, Redlands, CA).

GFP fluorescence was monitored using a Leica MZ16 stereomicroscope equipped with a mercury lamp, a Leica GFP2 filter set, and a Leica DFC420 C digital camera.

A solution of GA, MeJA or SA was added uniformly onto the surface of ICM plates using sterile glass beads. Two‐centimetre roots were then added and grown on each plate. After 1 day of growth on the plate, each root was treated with 400 juvenile HG 2.5.7‐type nematodes. For GA₃ treatments, Peking unedited roots were treated with four different concentrations of GA₃ (0, 0.1, 1 and 10 μm). One day later, 400 J2 HG 2.5.7‐type nematodes were added around the roots. An image of the whole root was captured 12 days after inoculation using a Leica MZ16 stereomicroscope with an attached camera. The length of first‐order fine roots and the diameters of the first‐order fine roots and main root were measured from at least 10 randomly selected roots. Measurements were performed using Adobe Illustrator. After imaging, the whole root was stained, and nematode development was quantified as described above. For MeJA treatment, DELLA‐edited roots and unedited roots both from Peking were placed on ICM plates with two MeJA concentration treatments (0, 150 μm) and inoculated with HG 2.5.7‐type SCN followed by a nematode demographics assay and the root architecture measurement as described above. For SA treatments, DELLA‐edited roots were placed on ICM plates with 1 mm SA and inoculated with HG 2.5.7‐type SCN followed by nematode demographics assays as described above.

The fixed samples were dehydrated in a graded series of ethanol (60%, 80%, 96%, and absolute ethanol) using a dehydration system with agitation and vacuum. The blocks were infiltrated with Kulzer Technovit 7200 VLC-resin. Infiltrated specimens were placed into embedding molds, and polymerization was performed under blue- and white light. Polymerized blocks were sectioned in a mesio-distal direction and parallel to the long axis of each implant. The slices were reduced by microgrinding and polishing using an Exakt grinding unit to an even thickness of 60–70 µm. Sections were stained with RBS and counter-stained with acid fuchsin and examined using both a Leica MZ16 stereomicroscope and a Leica 6000DRB light microscope. Histomorphometric measurements were performed by using software (ImageAcess, Imagic, Switzerland) to calculate the percentage of osteoid, mineralized new bone, and remaining old bone along the bone–implant contact surface. BIC was calculated based on the sum of osteoid, mineralized new bone, and remaining old bone.

Five specimens were examined from 46°59'45"S, 38°00'39"E, at depths of 360–376 m, between Marion and Prince Edward Island south of South Africa. The holotype was stained with methyl green for photography using a Canon EOS 7D camera with a Macro EF 100 mm lens and a Spot Flex CCD 15.2 fitted on a Leica MZ16 Stereo microscope at the Australian Museum, Sydney. The software Helicon Focus 5.3 was used for focus stacking. Another specimen from the same sample was dehydrated in ethanol, critical point dried, coated with 20 nm of gold and examined under a JEOL JSM-6480 Scanning Electron Microscope (SEM) at Macquarie University, Sydney. Terminology follows that of Hutchings et al. (2019). Data on the holotype are given, with the variations of the other material, all designated as paratypes, given in parentheses in the case of complete specimens. All material is deposited in Iziko Museums of South Africa (formerly South African Museum, Cape Town).

Mice were kept in a controlled 12-hour light/12-hour dark cycle, where the light cycle began at 7 am. Embryos were generated by timed mating between heterozygous animals, with noon on the day of finding a vaginal plug designated E0.5. To obtain E8.0 stage embryos, mice were housed using a reversed light/dark cycle, with the light cycle beginning at 10 pm. Embryos were dissected from the uterus in phosphate buffered saline (PBS) containing 10% newborn calf serum (Invitrogen), examined and photographed on a Leica MZ16 stereomicroscope. Embryos were scored as achieving Closure 1 when there was evidence of contact between the neural folds in the mid-region of the embryo, such that the neural folds could no longer be distinguished and a continuous surface ectoderm was observed.