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The specimen substrate was made from directionally solidified nickel base superalloy IN100 DS, and the coating system comprised a NiCoCrAlY bond coat and a ceramic top coat from partially stabilized zirconia.
The concentration of the cell seeded onto the specimen substrate was 2 × 104 cells per well.
The frozen specimen substrate was then retracted into the specimen isolation chamber and transferred directly onto the cryo-stage within the FIB/SEM.
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A 200 nm TiN-coated specimen with the same Ti6Al4V substrate was also built by PVD as a benchmark.
It was observed that on the non-boiled specimens, the contact between splat and substrate was poor, possibly due to the desorption of adsorbates/condensates present on the substrate surface, and no splats were formed on the boiled specimens, due to the presence of a thick hydroxide layer.
The substrate was then transferred out of the FIB/SEM using the customized specimen transfer shuttle device and loaded onto the liquid nitrogen-cooled specimen carousel of the ETH (Fig. 1d).
In all specimens, the substrate/coating interface was sharp and no Ti interlayer or modification of the substrate was observed.
The detection substrate was nitrophenylphosphatate (Sigma).
Cleavage of the substrate was measured using a microtiter plate reader (absorbance 405 nm) and normalized to the weight of the biopsy specimens used in each specimen.
Specimens of each substrate were prepared for microtensile bond strength test/μTBS (dentin and composite) or shear/SBS test (enamel and porcelain).
Perhaps the variance of different specimens of the same substrate is much larger than the difference between the 'with' and 'without' moth conditions?
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