Figure 4.

Key experimental factors in designing HST experiments include: (1) the choice of tissue area, (2) the number and sizes of fields of view (FoVs), and (3) the number of cells and spots. These experimental factors can affect the statistical power needed to achieve the research goals, e.g., those mentioned in Figure 2. For example, the choice of tissue area, along with the number and sizes of FoVs, can determine the degree to which the biological aspects of our interest (e.g., interesting cell subpopulations, or cell-cell communications) are captured in the generated HST data. Likewise, the number of cells and spots can affect the signal-to-noise ratios (effect sizes) of the generated HST data.