Cells grown in the Lab-Tek II chamber slide (Thermo ScientificTM, 154941) were fixed with 5% glutaraldehyde (Sigma-Aldrich, G5582) for 2 h at 4°C and washed with PBS for three times and proceeded for dehydration. Dehydration was performed in graded concentration of ethanol (Sigma-Aldrich, 1.07017) from 25%, 50%, 75%, 95% to 100% for 10 min and a final 10 min incubation with 100% acetone (Sigma-Aldrich, 179124). Cells are infiltrated in 100% acetone: resin (1:6) overnight at room temperature and incubated at 80°C oven for 4 h. The embedded cells were trimmed, sectioned, and imagined by TEM (JEOL 1400FLASH TEM).
1400 flash tem
Manufactured by JEOL
The JEOL 1400-Flash TEM is a transmission electron microscope (TEM) that provides high-resolution imaging capabilities. The core function of the 1400-Flash TEM is to generate detailed images of specimens by transmitting a beam of electrons through a thin sample, allowing for the examination of the internal structure and composition of materials at the nanoscale level.
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Lab products found in correlation
4 protocols using 1400 flash tem
1
Glutaraldehyde Fixation and Embedding for TEM
2
Ultrastructural Analysis of B. distachyon Leaves
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The middle third of fully expanded leaves from 3-week-old B. distachyon plants were collected for TEM analysis. Samples (1 mm2 size) were fixed, stained, and sectioned as described in Lundquist et al. (2013) (link). The images were captured by a JEOL 1400 Flash TEM. Measurements of plastoglobule densities in chloroplast areas were made using ImageJ.
3
Nanoemulsion Surface Morphology Analysis
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Surface morphology of the optimized CIP-NE formulation was observed through TEM [55 (link),66 (link)]. The sample was analyzed through negative staining protocol. The grid is placed over a 20 µL drop of the sample solution for 1 min. The excess sample is drawn-off the grid with the help of a filter paper. The grid is washed briefly by dipping it in distilled water and removing the excess water from grid by using filter paper. The grid is stained immediately using UranyLess for 1 min and then allowed to dry for a few minutes, followed by imaging using JEOL 1400-Flash TEM. All the images are taken at 25K times magnification.
4
Ultrastructural Imaging via EPON Embedding
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Samples were processed as described above with the exception that after the dehydration the samples were stepwise infiltrated with EPON resin (2 h 2:1, overnight 1:1, 3 h 1:2, 3 h 1:3 acetone:EPON, overnight 100% EPON) followed by embedding in freshly prepared EPON resin. After curing for 48 h at 60 °C the resin blocks were trimmed by hand using a razor blade. Ultrathin sections (50 nm) sectioned on a diamond knife (Diatome ultra 45) using a Reichert-Jung ultracut E ultramicrotome and mounted on copper grids (100# hexagonal) with a carbon coated formvar film and post-stained using 2% uranyl acetate and Reynolds lead citrate before imaging in a JEOL 1400-Flash TEM operating at 120 kV.
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