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Spleens were disaggregated through a 70 μm cell strainer into RPMI 1640.
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Nuclei were disaggregated then filtered through a 40 μm mesh prior to flow sorting with an Influx cytometer (Becton-Dickinson, San Jose, CA, USA) with ultraviolet excitation and DAPI emission collected at >450 nm.
Spleens were disaggregated by forcing tissue through a steel mesh and then filtered through a 40 μm filter to produce a single-cell suspension.
The tumor biopsies were disaggregated into single cells, then run through a Percoll gradient to enrich for cancer cells (methods).
Tumor cells were disaggregated by gentle pipetting then filtered through a 40 μm filter and kept on ice at 10 cells/mL.
Cells were disaggregated by vigorous pipetting and were passed through a 30-µm nylon mesh to remove any remaining clumps of tissue.
Dechorionated embryos were disaggregated on ice in 1×PBS and serially passed through 105- and 40-µm filters.
Nuclei were disaggregated subsequently with 20G and 25G needles and filtered through a 50- and a 30-μm mesh.
The pooled organoids were disaggregated by pipetting in 0.25% trypsin-EDTA (Sigma), followed by filtration through a 40μm mesh (Becton Dickinson, San Jose, CA, USA) to yield a predominantly single-cell suspension.
Next, EBs were disaggregated and re-plated in piPSCs growth conditions for 7 days.
Through an iterative process of coding and team discussion, superfluous codes were eliminated and overpopulated single themes were disaggregated as part of a second stage of coding.
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CEO of Professional Science Editing for Scientists @ prosciediting.com