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Grids were placed in 10 µl of virus sample for 1 minute, washed three times in sterile filtered water, and stained in 10 µl drops of 1% uranyl acetate three times, allowing the grids to soak in the last drop of stain for 2 min. After drying grids, samples were examined under an electron microscope (Phillips CM100) at 52K magnification.
A drop of stain solution was placed on each section and left to absorb for 1 min before washing the slides with sprayed water.
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After cooling, the cover slip was removed and two drops of stain were applied.
A coated grid with sample adsorbed to the surface is floated on a drop of negative stain for 0.5 2 min, excess stain wicked away with a piece of filter paper, air dried for 1 3 min, and examined by electron microscopy.
A total of 10 μL of peptide sample was pipetted on to grid and allowed to sit for 10 min. The excess was blotted away and finally a drop of PTA stain was added on to grid and allowed to sit for another 3 min. Excess stain was wicked away and grid was kept to dry overnight before TEM imaging.
A drop of the staining solution was added on to the film and the excess of the solution was drained off.
Mosquito guts were dissected on microscopic slides in several drops of the solution, mounted on other microscopic slides in a drop of the staining solution and covered with cover slips.
Staining of the filamentous actin cytoskeleton was carried out with TRITC conjugated phalloidin (Sigma), at a 1/500 dilution in 1X PBS by inverting the coverslip onto a 60 µl drop of the staining solution and incubating at room temperature in the dark for 1 h.
The sample was washed once with a drop of water, stained with a drop of 2% uranyl acetate and air dried.
Similarly for acridine orange staining, one drop of 0.1% the stain was added onto the section and a coverslip placed on it.
Subsequently, the cover slip was carefully separated from the agar, placed on a slide with a drop of cotton blue stain and observed under the microscope.
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