Fig. 1: Development of an erasable multiplexed-immunofluorescence method.
From: Precise immunofluorescence canceling for highly multiplexed imaging to capture specific cell states
a Schematic diagram of a fluorescence-cleavable antibody. b, c Immunofluorescence images of IMR90 cells stained with an anti-histone H3.1 PECAb or direct-labeled antibodies. The fluorescence signals were erased by TCEP. Scale bars, 10 µm in (b), 20 µm in (c). Quantified values were plotted in (c). Values are the mean ± SEM. Exact P values from one way ANOVA with Dunnett’s multiple comparisons test (n = 3 biologically independent experiments) are indicated in the panels and Source Data. d Top panels, immunofluorescence images of IMR90 cells stained with indicated antibodies. Scale bars, 10 µm. Bottom panels, the profile of fluorescence intensity on the yellow line drawn on the immunostaining images using an indicated full antibody set. e Schematics of SAHF staining. Top panel, induction of senescence. Bottom panel, PECAbs used to stain the IMR90 cell nuclei. Blue fill shows relevant features to the indicated antibody. f SeqIS images of IMR90 cells. Cells were sequentially stained with the antibodies. Areas within white dashed lines are magnified in the right panels. Scale bars for left: 5 µm, for magnified panels: 1 µm. Experiments in (d, f) were performed for 2 times and the representative images are shown. Source data are provided as a Source Data file.
