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Since cortical staining with voltage-sensitive dye will inevitably contain a certain degree of "patchiness," an ideal normalization method should be insensitive to the quality of staining.
Corresponding percentages of identified lymphocytes (sum of T + B + NK) to the percentage of gated lymphocytes was used to control the quality of staining.
Indeed, the reviewer is correct about the intracellular AcTUB staining, however, we do not believe this is due to poor quality of staining.
Negative (omission of antibody) and positive controls (oral cancer) cases were run to verify the quality of staining and confirmation of the procedures.
The cancer specimens were from resection and not from biopsy, which may have reduced a potential influence of the quality of staining for p65.
Correspondence of the percentage of identified lymphocytes (sum of T + B + NK cells) to the percentage of lymphocytes in the gate was used to control the quality of staining.
Similar(46)
The staining in negative controls was instead discriminated as non-specific based on the quality and location of staining.
***A case/control was labelled as positive if any positive signal was recorded in a lesion irrespective of the quality of stain.
Heterogeneous quality of the staining result may occur with CISH.
To ensure the quality of ZN staining the following quality control measures were taken.
These plots provide an objective diagnostic of staining quality highlighting slides for further investigation.
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CEO of Professional Science Editing for Scientists @ prosciediting.com