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After 3 washes in the same buffer, cells were postfixed in 1% osmium tetroxide in 0.1 M cacodylate buffer, pH 7.2, for 1 hour at room temperature.
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After washing with 10% sucrose in the same buffer, the cells were postfixed with 1% OsO4 in the same buffer and block-stained with 1% aqueous uranyl acetate.
After a rinse in 0.1 M phosphate buffer, the cells were postfixed in ferrocyanide-reduced osmium tetroxide (Karnovsky, 1971) for two hours, then rinsed in distilled water.
Fixation was carried out for 2 hr at 4°C; after several washings in cold phosphate buffer (0.1 M, pH 7.4) the cells were postfixed for 1 hr in 1% phosphate-buffered osmium tetroxide.
Cells were washed and fixed using a cold buffer containing 3% glutaraldehyde in 0.11 M cacodylate for 30 min. After rinsing with phosphate-buffered saline (PBS), the cells were postfixed in osmium tetroxide (1%) and embedded in Epon resin.
After washing, cells were postfixed in 0.1 M sodium cacodylate buffer containing 1% OsO4 for an additional 1 hour at 4°C.
The cells were postfixed with 1% osmium tetroxide in the same buffer for 30 min, then washed three times with PBS, dehydrated through a series of alcohol concentrations (30%, 50 %, 70 %, 90 %, 100 embedded in Epon, and sliced to a thickness of 70 nm.
Cells were postfixed using a 1%(v/v) formaldehyde solution in PBS for 10 min and washed again with PBS.
The cells were postfixed in 1% osmium tetroxide for 1 h at room temperature and washed.
After washing twice with 0.1 M PBS, the cells were postfixed with 1% osmium tetroxide at temperature for 1 h.
The cells were postfixed with OsO4, dehydrated and embedded in Quetol-812 (Nisshin EM, #340).
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