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One set of sections was stained with Nissl (cresyl violet).
One set of sections was stained with hematoxylin and eosin using routine procedures.
One set of sections was used for Nissl staining and another for ChAT immunocytochemistry.
One set of sections from each rat was stained using the Nissl method.
One set of sections was stained for Nissl substance (0.5% cresyl violet) and three other series were mounted for rhodamine and fluorescein visualization using two-photon microscopy.
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15 One set of the sections was incubated with primary polyclonal rabbit anti p-cPLA2 anti p-cPLA2100) overnight antibody
One set of the sections was stained with HE and the others were used for immunohistochemistry and for pathological diagnoses.
One set of the sections was stained for myelin with Luxol fast blue, and the other was counterstained with cresyl violet eosin.
For architectonically identifying V1 and its laminar structure, one set of brain sections was processed for cytochrome oxidase (CO) (Wong-Riley 1979) and another set was stained for Nissl substance.
From each of these animals, one set of coronal sections comprising the organum vasculosum of the lamina terminalis (OVLT) through the caudal border of the anterior ventral periventricular nucleus (AVPV) were immunolabeled for GnRH and Fos or Kiss using immunohistochemistry (IHC) methods described in detail elsewhere [ 4, 20, 48].
Every orexin-positive neuron was counted under a 63x oil objective lens throughout the thickness of the section in one set of the serial sections collected and the total number of orexin-positive neurons in the left and right hypothalamus was then estimated using a modified fractionator method [ 41– 41].
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