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The number of pericytes per EC (pericytes coverage) was calculated by manually counting the number of Cd31+ and Ng2+ cells in a field of view, and then counting how many were in contact with Cd31+ cells.
The total number of vehicles passing each of these locations was obtained by manually counting the vehicles in the video.
The surface coverage of AuNPs on electrode surface was estimated by manually counting the AuNPs amount in different analysis areas (1 μm × 1 μm) in SEM images and the average density of nanoparticles was adopted as coverage [20].
Spine density was calculated by manually counting the number of spines expressed over a 30 µm segment of dendrite.
Quantitation of Hsp70 labeling on exosomes was done by manually counting the number of gold label/vesicles on 20 systematically selected images per sample.
Finally, target genes of KD fibroblasts were normalized to those of WT. Cell proliferation was assessed by manually counting the number of viable cells.
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The number of neutrophils and macrophages comprising ME cellular infiltrates was assessed by two independent observers, by manually counting these cell types in five randomly selected clusters of cellular ME effusions for each ME in a 400x high power field.
Semi-quantification of CCL2- and CCR2-positive cell numbers was performed by unbiased manually counting the number of positive cells with a clearly visible cell nucleus within the region of interest (ROI).
To estimate adipocyte size, the number of adipocytes per mm was determined on scanned sections by manually counting all the adipocytes in an area of 4-5 mm, using Aperio ImageScope (Leica Biosystems Imaging Inc., CA, USA).
The number of total cells was determined by manually counting cells in the bright-field channel using the Fiji cell counter plugin.
Several kinds of data can be acquired by manually counting observations from the video record, such as traffic volume, quantity of curb parking and quantity of traffic conflicts.
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