TEM was conducted to investigate AuNP internalization in BeWo cells. Therefore, cells were cultivated on inserts for 3 days as described above and treated with 50 µg/mL Au-4-COONa or 25 µg/mL Au-3-PEG for 24 h at 37 °C and 5% CO2. Cells were detached from 11 inserts per condition (without NPs, Au-3-PEG, Au-4-COONa) using 0.5% Trypsin–EDTA, pelleted by centrifugation (200 g, 5 min) and sucked up into a capillary tube (Leica-Microsystems). Cells enriched in the capillaries were immediately fixed in 3% glutaraldehyde in 0.1 M sodium cacodylate buffer and washed in 0.2 M sodium cacodylate buffer. After a post-fixation step in 2% osmium tetroxide in 0.1 M sodium cacodylate buffer, samples were dehydrated through a graded ethanol series followed by acetone and finally embedded in Epon resin (Sigma-Aldrich, Buchs, Switzerland). Ultrathin sections were contrasted with 2% uranyl acetate and lead citrate (Reynolds 1963) before imaged in a Zeiss EM 900 (Carl Zeiss Microscopy GmbH, Germany) at 80 kV.
Capillary tube
Manufactured by Leica
A capillary tube is a small, narrow glass or plastic tube with a narrow internal diameter, typically between 0.1 and 1.1 millimeters. It is used to handle and transport small volumes of liquids through the process of capillary action.
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Lab products found in correlation
2 protocols using capillary tube
1
TEM Analysis of AuNP Internalization in BeWo Cells
2
Ultrastructural Analysis of Au-NP Treated A549 Cells
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A549 cells were seeded into 6-well plates at a density of 4 × 10 5 cells per well in a total volume of 2.5 ml complete cell culture medium. After 24 h, cells were treated with 100 μg ml -1 of Au-NP in 2.5 ml complete cell culture medium for 24 h. Cells were detached from wells using 0.5% Trypsin-EDTA, pelleted by centrifugation (200g, 5 min) and drawn up into a capillary tube (Leica-Microsystems). Therein, cells were fixed in 3% glutaraldehyde in 0.1 M sodium cacodylate buffer. After a post-fixation step in 2% osmium tetroxide in 0.1 M sodium cacodylate buffer, cells were dehydrated through a graded ethanol series, followed by acetone, and finally embedded in Epon resin (Sigma-Aldrich). Ultrathin sections were contrasted with 2% uranyl acetate and lead citrate 58 before observation in a Zeiss EM 900 TEM (Carl Zeiss Microscopy, GmbH, Germany) at 80 kV.
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