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Appropriate levels of gene expression were detected using antisense riboprobes, whereas hybridization with the riboprobe in the sense direction resulted in a complete lack of stain (Fig. 2).
Water channels have been indirectly indicated by CLSM methods, assuming that they correspond with voids, but CLSM easily overestimates the presence of voids, because they also appear due to lack of stain penetration.
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Lesions with areas of lack of staining were restained at two higher antibody concentrations.
The coordinates of the points on the cellular outline were extracted manually to obtain a smooth outline for each image extending from the bottom to the top of the cell until the outline was difficult to identify due to the lack of staining in the top region.
Viability was assessed by mobility and lack of staining after challenge with trypan blue.
Viability was assessed by mobility of the parasite flagellum and lack of staining after challenge with trypan blue.
The specificity of the IF assay was supported by the lack of staining seen in the pre-treatment biopsies in any dogs (Fig. 2A, B).
Specificity of antibody labelling was demonstrated by the lack of staining after substituting proper control immunoglobulins (Rabbit primary antibody isotype control, Invitrogen) for the primary antibodies.
The specifity of the antibody for δ-toxin is demonstrated by lack of staining of normal human keratinocytes in the absence of δ-toxin but positive staining in the presence of δ-toxin (Figure 1c).
The pyramidal neurons in the cornu ammonis (CA) and the polymorphic layer of the dentate gyrus exhibited particularly high levels of staining that contrast with the lack of staining in adjacent areas such as the molecular layer (Figure 2A).
Viable cells were distinguished by the lack of staining with propidium iodide (1 µg/ml) and considered positive for ALDH based on a control staining reaction using the enzyme inhibitor diethylaminobenzaldehyde (DEAB).
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