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To elucidate whether the 3D aspect of the channel walls is the important factor, or only the imposed linear one-dimensional (1D) directionality of cellular movement, we performed migration experiments with microcontact printed adhesive lines.
To confirm the inhibitory activity of S100A14 knockdown on cell movement, we performed a dual-color wound healing assay.
To estimate the velocity of endosomal movement, we performed time-sequential observations at 1-sec intervals for 30 sec by spinning-disc confocal laser microscopy.
To characterize the route of sucrose movement, we performed dye transport studies, examining elongated internodes at the stage when sucrose begins to significantly accumulate within stems.
Measurment of inter-trial spatial movement: We performed FFT-based correlation between two images, one is a target/template reference and the other is the current image data.
To ensure that the detected vibrations in hESC chromatin were not the result of nuclear or even whole cell movement, we performed control line scan experiments across the nuclear envelope to understand any baseline hESC dynamics.
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To quantify this chromatin movement, we perform an autocorrelation analysis of the derived track and reveal two discrete peaks of positive correlation between 0.01 and 0.1 s, which correspond to the characteristic time(s) at which the chromatin density region shifts back and forth along the line (10 to 100Hz).
To test whether Hipk1 plays a role in regulating these and other cell movements, we performed over-expression and loss-of-function phenotypic experiments in whole embryos.
To compare movement duration and deceleration time for the different types of movements we performed a repeated measure analysis of variance with gestational week (14th vs. 18th) and type of movement (uterine wall, self-directed, other-directed) as within-subjects factors.
To determine whether PPK-1 has a role in spindle movements, we performed ppk-1 RNAi) ppk-1 RNAitubulin strain and imaged the spindle under a spinning disk microscope.
Finally, in order to ensure that our analysis did not miss potential Z → A or A → Z movements, we performed an additional analysis that examined cases in which there were multiple potential daughter genes on different chromosomes and families in which the parent gene may be on the Z or the autosomes.
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