Samples were fixed immediately with 2.5% glutaraldehyde in 0.1 M sodium cacodylate buffer (both Serva) for 30 min at RT and stored at 4 °C. Postfixation was performed with 1% osmium tetroxide (Electron Microscopy Sciences) and 0.8% potassium ferrocyanide II (Roth) in 0.1 Mol/L cacodylate buffer for 1.5 h followed by the dehydration of the samples in a graded ethanol series and the embedding of the samples in Epon resin (Roth). Finally, ultrathin sections with a thickness of 70 nm were stained with uranyl acetate and lead citrate. Samples were examined using a Zeiss EM 906 electron microscope at 80-kV acceleration voltage (Carl Zeiss).
0.1 m sodium cacodylate buffer
Manufactured by Serva Electrophoresis
Sourced in Germany
0.1 M sodium cacodylate buffer is a commonly used buffer solution in various laboratory applications. It provides a consistent pH environment to maintain the stability and function of biological samples during analysis or experiments. The buffer solution has a pH of approximately 7.4, making it suitable for a wide range of applications where a physiologically relevant pH is required.
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Lab products found in correlation
Epon resin, by Carl Roth (1 mentions)
Potassium ferrocyanide 2, by Carl Roth (1 mentions)
Em 906 electron microscope, by Zeiss (1 mentions)
Osmium tetroxide, by Electron Microscopy Sciences (1 mentions)
Potassium ferrocyanide, by Merck Group (1 mentions)
Em906, by Zeiss (1 mentions)
Ultracut s ultramicrotome, by Leica (1 mentions)
Reynold s lead citrate, by Merck Group (1 mentions)
2 protocols using 0.1 m sodium cacodylate buffer
1
Ultrastructural Analysis of Biological Samples
2
Ultrastructural Analysis of Mitotane-Treated Cells
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For transmission electron microscopy, two resistant and two nonresistant clones were thawed and cultured to confluence without mitotane. Cells were seeded on a 6-well plate (2 × 106 cells per well). After 24 h, cells were treated with 50 µM mitotane or vehicle control (DMSO). After 72 h, cells were washed once with ice cold PBS and fixed with 2.5% glutaraldehyde in 0.1 M sodium cacodylate buffer (both from Serva, Heidelberg, Germany) at room temperature for 30 min. Afterwards, fixation buffer was changed, and cells were stored at 4°C for 2–14 days. Cells were postfixed in 1% OsO4 (Science Services, Munich, Germany) and 0.8% potassium ferrocyanide (Merck) in 0.1 M sodium cacodylate buffer for 1.5 h and then progressively dehydrated in ethanol, followed by embedding in Epon (Serva). Ultrathin sections (70 nm) were prepared using an Ultracut S Ultramicrotome (Leica), stained with uranyl acetate and Reynold’s lead citrate (Merck), and microphotographs were taken using an electron microscope EM906 (Carl Zeiss).
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