The PET thin film coated onto a microscope slide was visually inspected using a polarized light microscope (Nikon Eclipse LV100N POL). Optical micrographs were taken using a Nikon DS-Fi2 camera through the NIS Elements imaging software (v 4.30). The PET-coated substrates were observed after the drying process to evaluate the homogeneity of the PET surface. This confirms that a uniform PET coating was achieved (Figure S1 in Supporting Information ).
Ds fi2
Manufactured by National Instruments
The DS-Fi2 is a digital camera module designed for microscopy and imaging applications. It features a 5.0-megapixel CMOS sensor with a maximum resolution of 2592 x 1944 pixels. The camera supports various exposure and gain controls, as well as color adjustment options. It connects to a computer or other device via a USB 2.0 interface.
Automatically generated - may contain errors
Lab products found in correlation
Eclipse ts100, by Nikon (2 mentions)
Eclipse lv100n pol, by Nikon (1 mentions)
Smz18 microscope, by Nikon (1 mentions)
Matrigel basement membrane matrix growth factor reduced, by BD (1 mentions)
Eclipse ti u, by Nikon (1 mentions)
Fluorescent brightener 28, by Merck Group (1 mentions)
D 4.50.00 64 bit, by National Instruments (1 mentions)
6 protocols using ds fi2
1
Uniform PET Thin Film Characterization
2
Nodal Loss-of-Function Phenotype Assay
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Aliquots of 25 pg of either lft1-gfp RNA or lft1-ΔDLE-gfp RNA were co-injected with rhodamine dextran (injection control) into 1 cell stage AB wild-type embryos. GFP and rhodamine signals were analysed for a subset of injected embryos. The embryos were incubated at 28°C until 2.5 days post fertilization and imaged using a Nikon SMZ18 microscope equipped with a Nikon DS-Fi2 color camera controlled by NIS-Elements F 4.0 software. Imaged embryos were scored by the following categories of Nodal loss-of-function phenotypes: wild type, mild, moderate, or severe. Phenotypes were independently scored by two researchers.
3
In Vitro Angiogenesis Assay
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Capillary-like endothelial tube formation was evaluated by an angiogenesis in vitro angiogenesis assay. 130 μL Matrigel basement membrane matrix growth factor reduced (BD Biosciences) was added to each well of precooled 24-well tissue culture plates. Pipette tips and Matrigel solution were kept cold during the procedure to avoid solidification. The plates were incubated for 1 h at 37°C to allow matrix solution to solidify. 4 × 104 cells in a final volume of 500 μL EBM-2 were seeded onto the surface of each well containing the polymerized matrix. Cells were pretreated with the pharmacological inhibitors indicated or with vehicle alone and stimulated with the specific agonists 100 ng/mL Ang-1 or 200 ng/mL Ang-2 for 4-5 hours at 37°C. Tube formation was inspected under an inverted microscope (Nikon Eclipse TS100) at 20x magnification and images were acquired by a digital camera (Nikon Ds Fi2, Nis elements F 4.00.00 software). The closed polygons formed in five random microscopic fields per well were counted and values averaged.
4
Quantification of Harmful Algal Blooms
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The samples (1 L) were taken manually in clean polyethylene bottles at a depth of 1, 7, and 14 m from the water surface and fixed with Lugol’s iodine solution in order to preserve them for further analysis, carried out at NRL on Marine Biotoxins (Cesenatico) according to the EU reference method UNI EN ISO 15204:2006 (Utermöhl’s method, 1958) [42 ]. The PSTs producing microalgal species were counted using settling chambers (25–50 mL) under an inverted Nikon microscope (Eclipse Ti- U, objectives: 20× CFI planapo, 40× CFI planapo, and 100× CFI planfluor oil) equipped with digital cameras (Nikon DSFi2) and NIS-Elements imaging software (version n. 4.20.00). The morphology of fixed cells was also analyzed with an Ultraviolet 100 W Mercury lamp. Plate patterns were studied according to Balech [43 ] after staining with Calcofluor white (Fluorescent Brightener 28, Sigma, Steinheim, Germany) and was observed under UV epifluorescence (the UV filter arrangement was for 330–380 nm excitation and 420 nm emission wavelength). Microalgal abundance was expressed as cell numbers per liter (cells/L). The quantitative detection limit for Alexandrium species count by Utermöhl’s method was 120 cells/L for 25 mL subsamples or 60 cell/L for 50 mL subsamples, and the level of significance was 0.05 for both.
5
Transmission Electron Microscopy of Developing Worms
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The measurements, images, and videos of live larvae were taken on a Nikon Eclipse TS100 inverted microscope equipped with a Nikon DS-Fi2 camera controlled by the NIS Elements version 4.3 Windows based imaging program.
For transmission electron microscopic analyses of the developing worms, samples of OvL4 (n = 10) on days 48–50, molting OvL4 (n = 10) on days 50–60, OvL5 (n = 10) on days 60–75 or 120 were fixed with 2.5% glutaraldehyde and 2% paraformaldehyde in sodium cacodylate buffer (0.1M) for 2 h. After fixation, worms were washed three times in sodium cacodylate buffer (0.1M) and post-fixed with 1.5% OsO4 for 1 h. Worms were then washed and dehydrated using a series of increasing ethanol concentrations (50–100%) with a final wash with propylene oxide. Samples were then embedded in plastic resin (Epon 812, EMS USA) and prepared for sectioning. Ultrathin sections were contrasted with UranyLess (EMS, USA) and lead citrate and then were analyzed under the Tecnai G2 Spirit transmission electron microscope.
For transmission electron microscopic analyses of the developing worms, samples of OvL4 (n = 10) on days 48–50, molting OvL4 (n = 10) on days 50–60, OvL5 (n = 10) on days 60–75 or 120 were fixed with 2.5% glutaraldehyde and 2% paraformaldehyde in sodium cacodylate buffer (0.1M) for 2 h. After fixation, worms were washed three times in sodium cacodylate buffer (0.1M) and post-fixed with 1.5% OsO4 for 1 h. Worms were then washed and dehydrated using a series of increasing ethanol concentrations (50–100%) with a final wash with propylene oxide. Samples were then embedded in plastic resin (Epon 812, EMS USA) and prepared for sectioning. Ultrathin sections were contrasted with UranyLess (EMS, USA) and lead citrate and then were analyzed under the Tecnai G2 Spirit transmission electron microscope.
6
Morphological Analysis of Aphid Embryos
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After the end of each experiment, females from each generation tested were collected directly from their host plants and placed into Eppendorf tubes containing 2.5% glutaraldehyde for further morphological investigations. All embryos were dissected from whole insects (n = 5 for each generation and for each amine), treated with tris-buffered saline and stored in glycerol, examined using a Nikon SMZ 25 stereoscopic microscope, photographed using a Nikon DS-Fi2 camera and measured with NIS-Elements D 4.50.00 64-Bit. The length of embryos and the length of the largest mature embryo were measured for each dissected female. The total number of embryos and the ratio of the number of mature to immature embryos were recorded. Embryos were considered as mature if the eyes of the embryo were pigmented. In addition, before dissection the size of all viviparous females from the control aphids and those treated with amines were measured.
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