Extended data Fig. 9: Multiplexed uncaging of cells in different areas.
From: Single-cell analysis of regions of interest (SCARI) using a photosensitive tag
(a) Schematic representation of multiplexed analysis of cells in adjacent areas. (b) Representative confocal image of OVCAR5 tumor cell culture consisting of adjacent islands of GFP+ cells (population 1) and GFP− cells stained for expression of a cell membrane marker (population 2). Scale bar represents 250μm. Enlarged areas are indicated by the boxes. Tumor cells in a first island (area A, marked with the dashed white line), containing cells from population 1, were uncaged in a first uncaging round and in situ stained with a first fluorescently labeled αFLAG antibody (αFLAG1). Subsequently, tumor cells in a second island in the same culture (area B, marked by the dashed yellow line), containing cells from population 2, were uncaged in a second round of uncaging. After staining with a second fluorescent αFLAG antibody (αFLAG2), flow cytometric analysis was performed. Images are representative of 2 independent experiments. (c) Bar plot shows the fraction of population 1 (GFP+) and population 2 (membrane-stained) cells within the AF594low and αFLAG1−high (that is uncaged in round 1) population, or within the AF594low and αFLAG2−high (that is uncaged in round 2) population (n = 3 technical replicates). Data are representative of 2 independent experiments.
