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If background structures such as horizontal cells or vessels are present in the samples, it may happen that parts of them will be counted for cells as well.
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For quantification of p24 staining, ten 400X fields were counted for cells with definitive cytoplasmic staining.
Random fields were counted for cells that had invaded the membrane, using an optics microscope.
All samples were counted for cell number in quadruplicate and diluted in 1 x HEPES buffer to the desired cell number (ranging from 3000 to 200 cells) in a final volume of 50 µl.
The number of EGFP-LC3 puncta per cell was counted for 50 cells.
Resuspended cells were counted for viable cell number using an NC-100 Nucleocounter (Chemometec, DK-3450 Allerod, Denmark), and then mixed in suspension with alginate for encapsulation.
At least 3000 cells were counted for each cell line across 3 coverslips.
An average of ∼300 350 cells were counted for each cell line in total.
A vertex was defined as the point where three cells meet, and the number of vertices was counted for individual cells.
All stained vesicles were counted for 60 cells for each condition at each time point.
Namely, tumor cells per unit area were counted for each cell type in each patient.
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