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As the measured TAMRA fluorescence intensity of spindles varied from extract to extract, we used the total average of TAMRA intensities from spindle structures in all extracts divided by the average TAMRA intensity of spindle structures per extract to correct the measured TAMRA values of each spindle in order to be able to compare experiments performed in different extracts.
(F ) Frequency of each spindle class among the progeny of XXX wild-type mothers.
The contour length of each spindle is noted in the right-hand column.
The pole-to-pole length of each spindle is noted in column to the right of each plot.
The simulations were designed to reproduce the morphology and biophysical characteristics of each spindle sufficiently closely to estimate the spindle's critical force.
Black arrowheads mark the three anaphase B spindles depicted in B. Circle-and-stick diagrams on the right represent cross-sections near to the poles and midzone of each spindle.
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We also used a number of different parameters in Imaris to calculate spindle volume, in case that was the source of variability, and we also performed a set of calculations where we manually set a region of interest around each spindle by eye, to rule out the possibility that the program was not accurately calculating spindle volume.
Diagrams on the left show the contour length and number of microtubules within each spindle.
We next considered whether the number and length of microtubules within each spindle maximise the spindles' critical force.
The lack of staining of a narrow band within each spindle suggested the presence of a midzone that excluded anti-tubulin antibodies (Fig. 1C; a and d, arrows) [27].
To test this hypothesis, we used a spindle tilt assay to measure the angle of each mitotic spindle relative to the substratum ((Delaval et al., 2011; Hehnly and Doxsey, 2014) Figure 7A).
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