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Proliferation was assessed by proliferating cell nuclear antigen (PCNA) staining along with counting of cells harvested from confluent monolayer.
Three methods of measuring cell proliferation, viz., cellular H-thymidine uptake, counting of cells in suspension, and counting of colonies of cells grown in agar contained in glass capillaries, were compared by studying cell growth kinetics using the L 1210 cell line.
The number of cells at the lower side of each membrane was quantified by counting of cells from three randomly chosen sections (3.7 mm2) using an Olympus IX 1 fluorescence microscope (Olympus, Planegg, Germany).
Apoptosis induction was quantified by counting of cells with a characteristic apoptotic morphology after DNA staining with Hoechst 33342.
The percentage of Ki-67 labeled cells was determined after counting of cells in random, 200× microscopic fields.
Trypan blue stain, in which blue dye incorporating cells were scored as being dead was performed by counting of cells using a light microscope and a hemacytometer.
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Microscopy counts of cells with undetectable levels for each of the three proteins showed an average of 8% invasion compared to control treatments (Figure 3d spectrin and S22 adducin/p4.1).
Microscopy counts of cells with undetectable levels of spectrin/adducin/p4.1 showed 17%/15% /6%invasion compared to control treatments (Fig 4c spectrin and S24 adducin/p4.1).
To enable counting of cell clusters, cells were examined under 400× and 1000× magnifications.
Cell growth was monitored by daily counting of cell numbers with a hemacytometer.
Colorization and manual counting of cell numbers were performed using Metamorph (Universal Imaging, Sunnyvale, CA).
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