Smz800 stereomicroscope

Grass leaves (two grams/dish) were dipped in LC₃₀ (18.2 ppm) and LC₅₀ (27.7 ppm) concentrations of Imunit insecticide solution for 10 s. Distilled water served as a control. The treated leaves were air-dried for one hour at room temperature and placed in plastic Petri dishes (12 cm in diameter) with a hole (2 cm in diameter) capped with a muslin cloth. Then, 60 third-instar larvae were released separately into each Petri dish and kept under experimental conditions for 48 h. After 48 h, the surviving larvae were separately transferred to untreated leaves in plastic Petri dishes. The Petri dishes were checked daily until pupation, and developmental time was recorded for all immature stages sing a Nikon SMZ800 stereomicroscope (Japan). After the moth imago emergence, females were coupled with males obtained from the selfsame treatment in separate plastic containers (11 cm length × 9 cm width × 4 cm height) containing 20% honey solution as food. The adults were transferred to new containers every day, and the total number of eggs laid was counted daily. This procedure was continued until all the adults died. All experiments were performed in the growth chamber set at 25 ± 1 °C, 60 ± 5% RH, and 16:8 h (L:D) photoperiod.

Fertilized chicken eggs (EARL Les Bruyères, FR) (less sentient model according to 3R rule) were incubated in egg-racks at 37 °C and a 65% humidified atmosphere in a rotatory incubator allowing rotations of 25 degrees every 6 h. On day 3 of embryonic development, a window was made in the eggshell and sealed with adhesive film (Durapore tape). On embryonic day 10, once the CAM was fully formed, a gentle laceration of its surface was performed using a scalpel, and plastic rings (made from Nunc Thermanox coverslips) were put on the surface of the CAM [21 (link)]. Then, 4 × 106 of shLRP-1 or shCtrl MDA MB-231 cells were deposited as a thin layer on the surface of the fertilized chicken eggs. On day 17, CAMs were fixed in vivo (4% paraformaldehyde, RT, 30 min) and included. Digital photographs were taken under a Nikon SMZ800 stereomicroscope. A quantitative analysis was performed using a MATLAB routine developed by Dr El-Hadi Djermoune [22 (link)] in which the vascular structures’ segmentation leaned on the vesselness probability map of the 2D images [23 (link)].

Forty adult A. amphalodes were collected at Amanzi in order to investigate the prey-capture behaviour of the spider. After transfer to the laboratory, adult spiders of both sexes were placed separately into an arena consisting of 250 ml plastic bottles (8 cm in diameter, 15 cm tall) containing a 3 cm layer of sand in the bottom. They were left for three days to settle down at room temperature (~25 °C) and a natural LD (12:12) regime, during which time they were starved. Spiders usually dug themselves into the sand immediately after being released into the arena. Hodotermes mossambicus termites from one nest were collected in a suburban grassland in Langenhoven Park, Bloemfontein and kept together in plastic containers (40 cm in diameter) filled with soil. Trials began when a termite was introduced to the arena occupied by a spider. The hunting sequence was observed and recorded using handycam Canon Legria HF G10. If the spider did not start hunting within one hour, the prey was removed. We measured the paralysis latency as the time between successful attack and termite immobilization. After the trial the whole body length of termites and prosoma length of the spider were measured with a Nikon SMZ800 stereomicroscope.

Culture and morphological observations:
A mold isolate was incubated in petri dishes with potato dextrose agar (PDA; Becton Dickinson, USA) at 15 °C for 1–3 weeks in dark condition. Microscopic slides were prepared from the portions of the colonies grown on the PDA plates by mounting them in lactophenol (with/without cotton blue). Microscopic examinations were performed with a SMZ800 stereomicroscope (Nikon, Tokyo, Japan) and a BX51 microscope (Olympus, Tokyo, Japan) with Nomarski interference contrast at magnifications of up to × 1500. All micrographs were captured with a digital camera (DS-Fi2-L3; Nikon, Tokyo, Japan).

A behavioral scan of each colony was completed once each day for the duration of the
month-long study by recording the instantaneous behavior and location observed for
every visible paint-marked worker. Each behavioral scan was performed at 20×
magnification with a Nikon SMZ800 stereomicroscope. We recorded 30 distinct
behaviors, but only 15 were observed more than 15 total times during the study period
(Supplementary file
1). We defined an individual as foraging if it was observed on a food or
water source or actually carrying food (i.e., foraging included the behaviors
‘on honey’, ‘on liver’, ‘on water’, or
‘carrying food’; Supplementary file 1). Each experimental colony contained four
identifiable locations that were redefined prior to each behavioral scan: brood area,
brood periphery, remaining nest area, and foraging area. The brood area was defined
as the central area within the nest containing all brood and queens (Edwards, 1991 (link)). The nest periphery was defined
as the region directly adjacent to the brood area, where workers were dense in
aggregation but not in contact with any of the brood. The nest area was defined as
the sparsely occupied remainder of the space within the nest, not including the brood
area and nest periphery. The foraging area included all areas outside of the nest.
Analyses of behavioral data were conducted in R (www.r-project.org).

CHV-1 strain V777 [8 (link)] was generously provided by Dr. Andrew Davison (University of Glasgow). Virus stocks were prepared and titrated by plaque assay on MDCK cells essentially as described previously [7 (link)]. MDCK cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) containing 8% fetal bovine serum (FBS), 50 U/mL penicillin, and 50 µg/mL of streptomycin. Cells were propagated at 37 °C in a humidified incubator with 5% CO₂. For the plaque assays, cells were plated in 12-well plates, and the next day the confluent monolayers were infected with a serial dilution of the virus stock in duplicate, and then overlayed with DMEM containing 2% FBS and 0.4% of methylcellulose. Three days post-infection, the cells were fixed and stained with crystal violet. Plaques were visualized using a Nikon SMZ800 stereomicroscope and counted manually.