Sc50 camera

Cytospin preparations of adult blood were stained with Giemsa (Sigma, St. Louis, MO, USA), imaged using a Leica DM E microscope with a 100× oil objective (NA = 1.25) and an Olympus SC50 camera and differential counts performed. Zebrafish that developed lymphocytic leukemia were fixed in formalin, followed by paraffin embedding, sectioning and staining with Hematoxylin and Eosin and imaged using an Olympus BX46 clinical microscope with a UPLFLN (PH) Plan Semi Apochromat (OFN 26.5) and DP22 camera, utilizing Olympus CellSens Entry 3.2 (Build 23706) software.

Mouse colons were flushed with cold PBS to remove all contents, and the entire colon was rolled into Swiss rolls. The tissues were fixed in 2% PFA for 12 h at 4 °C and then washed in 1x PBS and transferred to 70% ethanol until embedding. Swiss rolls were embedded in the same manner as described above for segments of mouse colon. Paraffin-embedded, PFA fixed tissues were sectioned at 4.5 μm. Tissue sections were de-paraffinized and hematoxylin and eosin stained (Mayer’s Hematoxylin, Thermo Scientific; Eosin 1%, Morphisto, Germany). Histopathological analyses were performed using a semi-quantitative scoring system^{78 (link)} that evaluated the severity of crypt damage and cellular infiltration, epithelial erosion and tissue thickening using a severity score from 0 to 3 (0 = intact, 1 = mild, 2 = moderate, 3 = severe), and those scores were multiplied by a score for percent involvement (0 = 0%, 1 = 1–25%, 2 = 26–50%, 3 = 50–100%). A trained and blinded scientist performed the scoring. Representative images were acquired using an Olympus CKX53 microscope and Olympus SC50 camera.

Cytospin preparations were prepared from embryonic and adult blood as well as adult kidney and stained with Giemsa (Sigma), and differential counts performed. These were imaged on a Leica DM E microscope with a 100 × oil objective (NA = 1.25) with an Olympus SC50 camera Alternatively, adult zebrafish kidney cells were prepared in ice-cold phosphate-buffered saline supplemented with 2 mM EDTA and 2% (v/v) fetal calf serum and passaged through a 40 μm sieve and analyzed using a FACSCantoII with cell populations identified in a side-scatter (SSC)/forward-scatter (FSC) plot as described [4 (link)]. Data were analyzed for significance with a Student’s t test.

T24 and TCCSUP cells were obtained from the American Type Culture Collection (ATCC) and cultured in Dulbecco’s modified Eagle’s medium (D6429, Sigma-Aldrich) with 10% fetal bovine serum (F0804, Sigma-Aldrich) and 100 U of penicillin/streptomycin (P0781, Sigma-Aldrich) in a humidified incubator at 37 °C in a 5% CO₂ atmosphere. Trypsin-EDTA (T4049; Sigma-Aldrich) was used for cell passage and division. For nerolidol treatment, nerolidol was mixed with the complete growth medium to obtain the indicated concentrations. For experiments involving the use of inhibitors, cells were treated with inhibitors dissolved in complete growth medium at the indicated concentrations for 1 h before nerolidol administration: H-89 (50 nM), z-Vad (20 or 40 μM), SB203580 (0.5, or 4 μM), DRN (10 μM), XSC (380 nM), KH7 (3 and 10 μM), and L755507 (50 pM or 600 nM). All chemicals were mixed with complete growth medium and treatments were applied by replacing the existing growth medium to ensure homogenous administration. For each treatment, an appropriate control was used, which was an equivalent amount of vehicle (usually DMSO). Microscopy was performed using an Olympus CHX41 microscope (Olympus Corporation, Tokyo, Japan) and cell photographs were taken using an SC50 camera (Olympus Corporation).

A segment of the left lung was fixed in Histofix (Histolab) and embedded into paraffin and sectioned (3 µm) with a microtome. The tissue sections were placed on slides (Superfrost Plus; Fisher Scientific) and deparaffinized in serial baths of xylene and ethanol followed by staining using Mayer hematoxylin and 0.2% eosin (Histolab) or picrosirius red staining kit (Abcam, Cambridge, United Kingdom). The stained slides were imaged using an Olympus BX60F microscope with an SC50 camera.

All samples were analyzed by an Olympus BX51 polarized optical microscope coupled to a Linkam hot-stage that uses nitrogen to control the temperature and manages the cooling rate. An Olympus SC50 camera linked to the microscope was employed to observe the samples and take micrographs. Samples were dissolved in acetone or cyclohexane, and drops of the solutions were placed on a glass substrate and dried at room temperature.

The specimens have been collected in the Dominican Republic during a field trip in summer 2021.
Nymphs were preserved in 100% ethanol. Nymphal habitus were photographed using a Canon EOS 6D camera and the Visionary Digital Passport imaging system (formerly available and distributed by Dun Inc., Virginia) and processed with Adobe Photoshop Lightroom and Helicon Focus version 5.3.
Two nymphs were dissected in Cellosolve (2-Ethoxyethanol) with subsequent embedding in Euparal medium and mounting on slides. Fore- and hind wingpads of a submature female nymph were dissected and subimaginal wings examined. Microscopic pictures were taken using an Olympus BX51 microscope coupled with an Olympus SC50 camera; photographs were enhanced with Olympus Stream Basic 2.3.2 stacking software and Adobe Photoshop version 21.2.2.
The material is deposited in the collections of the Museum of zoology, Lausanne (MZL) and the Museo National de Historia Natural "Prof. Eugenio de Jesús Marcano", Santo Domingo, Dominican Republic (MNHNSD).