Cotton leaves were sampled at 14 DAT and were subsequently sliced and prepared as described by Jiang et al. [62 (link)]. The leaf slices were subsequently examined using a transmission electron microscope (FEI Tecnai Spirit 120kv, FEI Company, Hillsborough, OR, USA).
Fei tecnai spirit 120kv
Manufactured by Thermo Fisher Scientific
Sourced in United States
The FEI Tecnai Spirit 120kV is a transmission electron microscope (TEM) that operates at an accelerating voltage of 120 kilovolts. It is designed to provide high-resolution imaging capabilities for a variety of sample types.
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4 protocols using fei tecnai spirit 120kv
1
Transmission Electron Microscopy of Cotton Leaves
2
Ultrastructural Analysis of Cfap70-KO Mouse Tissues
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Precipitation of sperm as well as tissues of testis and brain (∼1 mm3) from adult Cfap70-KO mice and their littermate wild-type mice were fixed with 2.5% (vol/vol) glutaraldehyde in 0.1 M phosphate buffer (PB) (pH 7.4), washed two times in PB and two times in ddH2O. Then, tissues were immersed in 1% (wt/vol) OsO4 and 1.5% (wt/vol) potassium ferricyanide aqueous solution at 4 °C for 2 h. After washing, the samples were dehydrated through graded alcohol (30%, 50%, 70%, 80%, 90%, 100%, 100%, 10 min each) into pure acetone (10 min twice). Samples were infiltrated in a graded mixture (3:1, 1:1, 1:3) of acetone and SPI-PON812 resin (21 mL SPO-PON812, 13 mL DDSA and 11 mL NMA), and the pure resin was changed. The specimens were embedded in pure resin with 1.5% BDMA and polymerized for 12 h at 45 °C and 48 h at 60 °C. The ultrathin sections (70 nm thick) were sectioned with a microtome (Leica EM UC6), double-stained with uranyl acetate and lead citrate, and examined by a transmission electron microscope (FEI Tecnai Spirit120kV). All reagents were purchased from Zhongjingkeyi Technology (Beijing, China).
3
Ultrastructural Analysis of WA Cells
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We assessed the surface morphological characteristics of WA using a scanning electron microscope (SEM) and observed the cellular ultrastructure using a transmission electron microscope (TEM). We collected the WA cells of the control, ethanol, and ethanol + Tre groups by centrifugation (4000 × g, 10 min), which were then rinsed 3 times with physiological saline. The cells were then suspended in 2.5% glutaraldehyde and underwent three washes with 0.1 M phosphate-buffered saline (PBS; pH 7.4). Subsequently, the cells were dehydrated with 0%, 50%, 70%, 85%, 95%, and 100% ethanol solutions. After drying with in a critical point dryer (Hitachi, Tokyo, Japan), WA cells were coated with a gold/palladium alloy, and then examined using an FEI Quanta FEG 450 SEM system (FEI, Hillsboro, OR, USA). The dehydrated WA cells were embedded in an epoxy resin and sectioned with a diamond knife (LKB-Nova, Elkin, NC, USA). The ultrathin sections were stained with lead citrate, viewed, and photographed with an FEI Tecnai spirit 120 kV (FEI).
4
Transmission Electron Microscopy Sample Preparation
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The cells were fixed with 2.5% (v/v) glutaraldehyde with phosphate buffer (PB) (0.1 M, pH 7.4) and then washed four times in PB. Then cells were postfixed with 1% (w/v) osmium tetraoxide for 2 h in PB at 4 °C, dehydrated through a graded ethanol series (30, 50, 70, 80, 90, 100, and 100%, 7 min each) into pure acetone (2 × 10 min). The samples were infiltrated in graded mixtures (3:1, 1:1, and 1:3) of acetone and SPI-PON812 resin (Beijing Zhongjingkeyi Technology Co., Ltd., Beijing, China) (16.2 g SPI-PON812, 10 g dodecanoyl succinic anhydride and 8.9 g N-methyl acrylamide), the changed pure resin. Finally, the cells were embedded in pure resin with 1.5% N, N-dimethylbenzylamine and polymerized for 12 h at 45 °C and for 48 h at 60 °C. The ultrathin sections (70 nm thick) were sectioned with a microtome (Leica EM UC6), double-stained with uranyl acetate and lead citrate and examined using transmission electron microscopy (FEI Tecnai Spirit 120kv, FEI Company, Hillsborough, OR, USA).
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