Population and/or single-cell snapshot data is nonambiguous and can yield many conclusions. (A) Cells can acquire multiple cell fates. (B) Single-cell snapshot analysis can reveal subpopulations otherwise masked in population averages. (C) Snapshot analysis is ambiguous and allows for alternative interpretations of the same data set. In this simple example, 4 input cells (white) give rise to 4 white cells and 4 red cells. Many more possibilities exist but are not displayed. (D) Alternative explanations also remain in clonal assays started from single cells. Here, a single cell input cell (white) generates 4 cells (red). Only some of many more possible alternative explanations are displayed. (E) Continuous quantification of fluorescence signals can reveal transient cellular states that would be missed by snapshot analysis. Here, a single input cell (white/red) gives rise to 4 white cells and 4 red cells expressing mutually exclusive lineage-promoting transcription factors. Differentiation into either cell type can occur directly or through a transient double-positive population. (F) Temporal deconvolution of continuously observed single-cell dynamics enables the undistorted analysis of cells at different developmental stages or cell cycle phases. Temporally aligned single-cell dynamics can reveal the sequence of molecular events prior to and after differentiation (indicated by red numbers).