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Particles of label were counted from three 1 μm2 areas from three area of the cell (pyrenoid and starch sheath, chloroplast excluding pyrenoid and starch sheath, outside of chloroplast) from three cells, for each antibody (APR and SiR) and negative control (secondary antibody only).
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During this loop, the number of cells that have changed their peak patch label is counted.
Only neurons with clear nuclei labeling were counted.
The number of cells expressing a PR and H-TdR (double-label) were counted in three random fields of view (400× magnification).
Only large and well-stained cells (with the whole body labeled) were counted.
Because the total number of calbindin-ir cells in this region was relatively small (< 100), sampling was not required; all cells that came into focus within the section and were moderately to darkly labeled were counted directly.
Touring, which the label is counting on the Girls to do, could also be a problem.
Two different images of each section were taken and the number of cells labeled was counted (n = 2 p73−/− compared to n = 2 control littermates).
The number of cells expressing an ER-α and H-TdR double-label was counted in three random fields of view (400× magnification).
Four random 20× images (each field with an average of 150 muscle fibers) were captured per muscle and the number of fibers with dysferlin labeling was counted and expressed as percent of total number of fibers.
DNA fibre spreading and labelling was carried out as described [3]. 100 doubly labelled fibres were counted for each condition.
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