Ckx41 inverted

Glass slide samples dyed with acridine orange were observed under a CKX41 inverted microscope (Olympus; 20x objective lens) equipped with LED fluorescence modules LED-FL-BG/MI and URFLT50 and a light source. The cells were observed under blue light and photographed using a microscopic digital camera DP80 equipped with the cellSens software (Olympus) set with a sensitivity of ISO 400 and an exposure time of 40 ms. Each slide was photographed over 50 times at various microscope fields of view, and each image was serially numbered and saved in the RGB-tiff format.

A total of 240 samples were screened under microscopy, for filamentous/colonial cyanobacteria and other phytoplankton. Water samples were preserved in 2% Lugol solution and stored in the dark at room temperature until further examination. Subsamples were transferred into sedimentation chambers (10 ml) for approximately 24 h before counting with an Olympus CKX 41 inverted light microscope. In each sample a minimum of 300 cells were counted (SD ≤ 11%). Phytoplankton cells including filamentous and colonial cyanobacteria were identified to genus and species level whenever possible. Morphological criteria for cyanobacteria were filament (trichome) length and shape, width and length of cells, presence of heterocysts, and presence and shape of akinetes. Taxonomical confirmation was achieved by consulting the database Nordic Microalgae¹ validated by the HELCOM Phytoplankton Expert Group. Cell biomass was calculated from biovolume (Olenina et al., 2006 ) and carbon content (Edler, 1979 ).

The differentiated 3T3-L1 adipocytes were fixed with 10% formalin, washed with 70% ethanol and phosphate-buffered saline (PBS), and finally stained with ORO solution. Stained lipid droplets were visualized using an Olympus CKX41 inverted microscopy (Olympus, Tokyo, Japan). We calculated the number of ORO stained lipid droplets for each group using Metamorph offline (Molecular Devices Co., Sunnyvale, CA). Cells were exposed to OYSGS for 8 days during adipocyte differentiation.

MDA-MB-231 cells (6.0 × 10⁵, 2.5 mL) suspended in culture media were seeded into a 6-well plate and allowed to grow to 80% confluence. Then, a straight scratch was made using a 200 μL pipette tip, and cells were washed followed by treatment with different concentrations of NLSPE5 in media with 0.5% FBS. Images were captured at 0 h, 16 h, 24 h, 40 h and 48 h after wounding using an Olympus CKX41 inverted microscope. The wound area was quantified using the ImageJ software (National Institutes of Health, (NIH), Bethesda, MD, USA), with a wound healing tool macro (Montpellier RIO Imaging, Montpellier, France). Treatment effects were described as the mean ± SE of three independent experiments.

The stage of the oestrous cycle was assessed daily by visual inspection28 (link) or from vaginal smears28 (link)29 (link). Oestrous smears were mounted on slides and imaged immediately using an Olympus CKX41 inverted microscope with a × 20/0.4 NA objective and a DP21 camera.