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The experimental results observed from the visualization cell are used to support the theoretically derived concept.
In each die design melt flow in the confluent region and die land to the die exit was observed through side windows of a visualization cell.
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To facilitate visualization, cells were assigned to one of nine broad planarian tissue-type classes (Fig. 2a), including both terminally differentiated cells and progenitors for those cells generated by neoblasts.
As for ceramide domain visualization, cells were incubated with antiCer, centrifuged at 1700 g for 10 min to remove supernatant (cells were resuspended with RBC buffer) and reincubated with Alexa Fluor 633 followed by an additional centrifugation step.
For actin and nuclei visualization, cells were cultured for 24 hours in 10% serum prior to cell staining and imaging.
For visualization, cells are positioned on a 2D lattice, where the coordinate of a cell is chosen according to its initial state of the genes x and y.
For cell morphology and actin cytoskeleton visualization, cells were fixed for 15 min in PBS supplemented with 4% formaldehyde, permeabilized with 0.1% Triton X-100 and stained with rhodamine-phalloidin (Molecular Probes) and DAPI (Sigma).
Prior to visualization, cells were concentrated 10-fold and 4′6-diamidino-2-phenylindole (DAPI) added to 0.02 mg/ml.
For nuclei visualization cells were incubated with Hoechst33342 (Sigma Aldrich) for 5 min and mounted with ProlongGold (Invitrogen).
For visualization cells were further incubated with fluorescence-labeled secondary antibody followed by analysis on LSM710 or LSM800 confocal microscopes (Zeiss).
For visualization, cells were washed, fixed, and incubated overnight at 37°C with X-gal chromogenic substrate at pH 6.0 as described [ 17].
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