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Sorted plates were immediately frozen on dry ice.
Sorted plates were immediately frozen on dry ice and stored at –80°C.
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LSK HSCs were FACS-sorted, plated for osteoclastogenesis assay at 2000 cells per well and stimulated with serial concentrations of M-CSF.
In vitro hematopoietic progenitor analysis was performed on dilutions of sorted cells plated in triplicates in methylcellulose medium (StemCell Technologies).
On page 6. "HNK-1+ve sorted cells plated on fibronectin or laminin surfaces showed neural immunoreactivity, including SoxE (recognizes Sox9 and Sox10).
The brightest GFP-positive cells were sorted and plated directly onto gelatin-coated dishes, ready for incubation with OSK lentiviral vectors (Supplementary Figure 2).
HNK-1 +ve sorted cells plated on fibronectin or laminin surfaces showed neural immunoreactivity, including SoxE (recognizes Sox9 and Sox10), HNK-1, HuC/D, Tuj1 and E-C8.
When sorted and plated separately, 5 to 10% of the CD49f+/EpCAM-/low SUM149PT cells differentiated into CD49f+/high/EpCAM+ basal-like cells, whereas the CD49f+/high/EpCAM+ basal-like cells maintained their differentiated status during in vitro culture.
The alternative option suggested by the reviewers involving the use of immunocytochemistry would be technically challenging given that freshly sorted and plated TGFP+ cells require a few hours to attach properly thus precluding antibody staining at the very start of the experiment.
The sorted cells were plated on 96-well plate for single cell colony selection.
Cells were infected at a multiplicity of infection (MOI) of 1.0 in the presence of acyclovir and at 10 h post-infection, either the GFP+ or GFP− cells were sorted by FACS, and the sorted cells were plated on fresh cells.
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