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Methods: We acquired high-resolution structural MRI and DSI of the cerebellum in four healthy female subjects at 3T. DSI tractography based on a streamline algorithm was performed to identify the circuits connecting the cerebellar cortex with the deep cerebellar nuclei, selected brainstem nuclei, and the thalamus.
We acquired high-resolution structural MRI and DSI of the cerebellum in four healthy female subjects at 3T. DSI tractography based on a streamline algorithm was performed to identify the circuits connecting the cerebellar cortex with the deep cerebellar nuclei, selected brainstem nuclei, and the thalamus.
See [ 2, 36] for details on the connectivity of nuclei selected for analysis.
We measured FRET in early anaphase cells displaying dumbbell-shaped nuclei selected from the 120 min time point of the experiment shown in Figure 2B.
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For each of the nuclei, select the connected components that form positive feedback loops.
To quantitate NPC, confocal images were taken on a Zeiss LSM ZeissLSM510 Confocal laser-scanning microscope using 63× oil objective (N/A 1.4), and the section with the widest cross-section of the nucleus selected for further analysis.
For each hESC nucleus selected (N = 10) three regions of chromatin were tested, by zooming in on a single chromatin density region and then scanning a line across it rapidly in time (see Experimental Procedures).
As with the hESC experiments, for each HELA nucleus selected (N = 10) we measured chromatin movement in three regions and produced for each a time series of the Hoechst 33342 intensity (chromatin density) along the line scan.
As with the hESC and HELA experiments, for each NIH3T3 nucleus selected (N = 10), we measured chromatin movement in three regions and produced for each a time series of the Hoechst 33342 intensity (chromatin density) along the line scan.
ImageJ was then used to calculate the mean gray value (average signal intensity over the selected area) and integrated density (mean gray value multiplied by the selected area) for the DAPI signal in each of the 18 25 slices in each nucleus selected for our study.
Areas of optimal tissue digestion and no overlapping nuclei were selected in each core for counting.
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