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Opaque zones were enumerated for each fish and width of the margin was noted.
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Opaque zone counts were converted to calendar ages based on timing of opaque zone completing and date of capture.
After ∼1 min, the white opaque zone indicative of the maturation stage became apparent.
The white opaque zone and remaining maturation-stage enamel were similarly dissected.
The presence of an opaque zone around a well indicated that the fibrinogen had been converted into fibrin.
Secretory-stage and transition-stage enamel samples, measuring 1.5 2 mm, were dissected apical to the white opaque zone.
A scalpel was used to dissect the developing enamel lying apical to the white opaque zone clear of the underlying dentine.
The adhering enamel organs were wiped away using damp paper tissue and the teeth allowed to air dry until the white opaque zone was visible.
Neither S. mutans cells nor culture supernatants converted fibrinogen into fibrin, while the positive control (thrombin) produced an opaque zone (data not shown).
The enamel organ was gently removed with a moist paper tissue and the tooth was air-dried for 1 min until the white opaque zone, representing the start of the maturation stage of amelogenesis, became visible (18).
One micro litre of colony materials from the opaque zone of the colonies was streaked on XLD + N and BG, and incubated at 37 ± 1°C for 24 ± 3 h.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com