Bx51m microscope

Attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) (Interspec 200-X Spectrophotometer, Interspectrum, Tõravere, Estonia) was used to characterize the samples before and after the addition of BPA.
To determine the structure and morphology of the eco-materials, Optical Microscopy (OM) (OLYMPUS BX51 M microscope, Tokyo, Japan) and Scanning electron microscopy (SEM) coupled with energy-dispersive spectra EDS (QUANTA 450 FEG microscope, Eindhoven, The Netherlands), equipped with a field emission gun and a 1.2 nm resolution X-ray energy dispersive spectrometer, with a resolution of 133 eV) were used.

CNU080 strain (GFP-Cys3) was incubated in several nutritional conditions depending on experimental design. Cells were grown overnight in 50 mL YEPD at 30 °C with 150 rpm rotation, wash with PBS three times, diluted to OD_600nm = 1 (50 mL), and incubated in various nutritional conditions at 30 °C for 1 and 2 h. A 100 µL aliquot was withdrawn for microscopy analysis. The cells were fixed with 4% formaldehyde (Sigma) (v/v) for 10 min at room temperature, washed twice with PBS and stained with 50 ηg/µL of DAPI (4′,6-Diamidine-2′-phenylindole dihydrochloride) for 10 min at room temperature. Cells were washed to remove the excess dye, and slides were prepared with 4 µL of ProLong antifade and 6 µL of the processed sample. Cells were viewed by direct fluorescent microscopy using an Olympus BX51M microscope and analysis was performed using Olympus CellSens, PhotoShop CS6, and ImageJ software. Percentages of nuclear GFP-Cys3 were obtained by counting the cells in which GFP and DAPI were overlaid (n > 100 cells/replicate). All microscopy experiments were done in triplicates.

AFM images were captured with Bruker Dimension Icon in ScanAsyst mode. The optical images were taken with an Olympus BX 51 M microscope. Raman and PL spectra were collected with Horiba-Jobin-Yvon Raman system under 532 nm laser excitation with a power of 2 mW. The Si peak at 520.7 cm⁻¹ was used for calibration. SHG measurements were conducted under an 820 nm femtosecond laser with excitation power of 0.4 mW (Spectra-Physics® Inspire™ ultrafast OPO system with tunable wavelength of 345–2500 nm, pulse duration of 100 fs and repetition rate of 80 MHz). Linearly polarized excitation light was focused on the sample by an Nikon objective (100X, N.A. = 0.95). SHG signal was detected with a spectrometer (Princeton Instruments, Acton SpectraPro® SP2500 PYLON). Atomically resolved high-angle annular dark-field STEM images were taken with an ARM-200CF (JEOL) transmission electron microscope operated at 200 kV and equipped with double spherical aberration (Cs) correctors. The attainable resolution of the probe defined by the objective pre-field is 78 picometers.

An Olympus BX51M microscope, with oculars at 10x fixed magnitude and a set of objectives with 5×, 10×, 20×, 50×, and 100× magnifications, equipped with a filter U25LBD Olympus for the white light and a UV source Olympus U-RFL-T, was used for image acquisition, supported with PrimoPlus software connected to the microscope by means of the Olympus DP70 scanner. The Ponceau S (3-hydroxy-4-(2-sulfo-4-[4-sulfophenylazo] phenylazo)-2,7-naphthalenedisulfonic acid sodium salt), a sodium salt of a diazo dye of a deep red color, was employed as the contrast marker to evaluate the penetration and migration on the adhesive into the mock-ups.

Optical microscopy (OM; BX51M microscope, Olympus, São Paulo, Brazil) and scanning electron microscopy (SEM; model EVO-LS15, Carl Zeiss, Jena, Germany) were used to reveal the microstructural characteristics. The samples were first placed through the usual metallographic processes, including SiC waterproof sandpaper grinding until #2000, polishing with colloidal alumina suspensions (0.25 m), and etching in Kroll’s solution.