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To ensure attachment when plating cells, dishes were treated with fibronectin (10 µg/ml) for 30 minutes then rinsed with warm, sterile PBS before adding cells.
The cells dishes were positioned on this phantom with 2 cm of a water-equivalent phantom above.
For Eed +/+, Eed −/−, and ES-ERT2 Ring1A −/− cells, dishes were coated with 0.1% gelatine and mitomycin-C inactivated primary embryonic fibroblasts (PEFs).
The cells dishes were placed on a water-equivalent phantom of 5 cm thickness to generate an enough back scattered radiation.
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After 3 h of infection, cell dishes were chilled on ice and rinsed twice with cold PBS.
HepG2.2.15 cells were grown on 10-cm cell dishes.
The uniformity of dose among cells in a dish is achieved by keeping the STD and the size of cell dish in certain ranges.
Those are precise position control of a cell dish holder, design of the cell dish, data acquisition of the microscopic image of a cell organelle (cell nucleus), data processing, reliable particle detection, software and hardware to integrate all these related data, and a system to control and irradiate a targeted spot with an exactly determined number of particles.
Those are precise position control of a cell dish holder, design of the cell dish, data acquisition of microscopic image of a cell organelle (cell nucleus), data processing, reliable particle detection, soft and hard wares to integrate all these related data and system to control and irradiate a targeted spot with exactly determined number of particles.
However, the inhomogeneous Am-241 distribution in a disc source did not affect the radial distribution of fluence rate at the inner bottom of cell dish when the dish is apart from the source sufficiently.
The treated cell dish was placed on the stage of a temperature-controlled microscope.
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