Cx43 light microscope

A total of 803 specimens of Syllinae were collected during August of 2014 in Bermuda by snorkelling or SCUBA diving in different substrates and depths (electronic supplementary material, table S2). Specimens were sorted at the Bermuda Institute of Ocean Sciences (BIOS) using a Nikon SM7 stereoscope and preserved in 96% ethanol for the subsequent molecular and morphological analyses. Further examination and identification were completed at Universidad Autónoma de Madrid (UAM) using an Olympus SZ 40 stereoscope and an Olympus CX 43 light microscope. Drawings were made to scale with a camera lucida attached to a Nikon Optiphot light microscope and from pictures taken with an Olympus CX41 light microscope. The width of specimens was measured at the level of the proventricle, excluding the parapodia.
For scanning electron microscopy, selected specimens were prepared on an Emitech K850 Critical Point Dryer, gold-coated with a Q150T-S Turbo-Pumper Sputter Coater and examined with a Hitachi S 3000N scanning electron microscopy (SEM) at the Interdepartmental Research Service (SIDI) of the UAM.
Vouchers for all newly collected specimens were deposited at the Museo Nacional de Ciencias Naturales (MNCN) in Madrid (Spain). Catalogue numbers, collection dates, locality and additional relevant information are listed in electronic supplementary material, table S2.

Four weeks after tumor implantation, nude mice were euthanized, and subcutaneous tumor tissues were collected. Concurrently, biopsy or postoperative samples of prostate cancer were obtained from clinical patients. The collected tissues were embedded in paraffin and sectioned into 4-µm slices. After deparaffinization using xylene, the sections underwent a series of ethanol washes (absolute ethanol, 95%, 85%, and 70% ethanol) for hydration. Subsequently, the sections were stained with hematoxylin for 10 min followed by eosin for 3 min. The resulting histological changes in prostate cancer tissues were observed under an Olympus CX-43 light microscope.

Transwell assay was conducted, as described previously [16 (link)]. Briefly, serum-free cell suspensions (2.5 × 10⁴ cells/mL) were made and 0.1 mL of the cell suspension was seeded to the top chamber of the transwell plates. Culture medium containing 10% FBS was added into the lower chamber. Cells were cultivated for 24 h at 37°C 5% CO₂. Membranes were cleaned using a cotton swab, followed by fixing with 4% polyformaldehyde (10 min). After washing twice, the top chamber was stained with 0.5% crystal violet (Sigma-Aldrich; St. Louis, USA) for 15 min at room temperature. The experiments were performed in triplicate. Cells were observed and counted under Olympus CX43 light microscope (×40 magnification). Then invasive cells in three fields were counted, and the average number was calculated.

Pancreatic tissue samples were collected, dissected, and immediately fixed in 10% formalin for 24 hours, dehydrated using a graded alcohol series, cleaned in xylene, and finally embedded in paraffin. Tissue sections were stained with hematoxylin and eosin (H&E) for histopathological analysis (23 ). All areas were viewed using an OLYMPUS CX43 light microscope using a 400x magnification and shot with an OLYMPUS SC52 camera. The area of the Islet of Langerhans was counted using ImageJ, and two blind histopathologists examined all histological anomalies. The histological state of the pancreas was evaluated and then compared across the various treatment groups for damage and regeneration of pancreatic islet cells.

Animals were sacrificed at the end of the experiment, and tissue samples from the pancreas and the adrenal glands were taken. These samples were then dissected, fixed immediately in 10% formalin for 24 h, dehydrated using a graded alcohol series, cleaned in xylene, and then embedded in paraffin. Sectioned tissues, 3 μm thick, were stained with hematoxylin and eosin (H&E) (35 ), for histopathological examination. All sections were examined using an OLYMPUS CX43 light microscope and photographed using an OLYMPUS SC52 camera from the microscopy unit of the Faculty of Veterinary Medicine, Sohag University, Department of Pathology and Clinical Pathology.