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Fig. 3 PhOTO-M Photoconversion Persists in Multiple Cell Types.
Zebrafish were photoconverted in a <100µm circular region of interest near the animal pole during gastrulation and were imaged the following day to visualize cell membranes at high resolution in different cell compartments. Even after multiple rounds of division, photoconverted membranes are still clearly visible above background. (A) Photoconverted membranes are clearly visible in the epithelium a day after photoconversion. Maximum intensity projections of a ventrally mounted zebrafish (anterior top) embryo 1 day post fertilization. Left to Right: Unconverted memb-Dendra2 (green), H2B-Cerulean (blue), photoconverted memb-Dendra2 (red) and a merged image of the three channels. (B) Photoconverted membranes are visible and are separated by unconverted membranes in the eye a day after photoconversion. Single focal plane images of the developing eye in a ventrally mounted zebrafish (anterior top) embryo 1 day post fertilization. Left to Right: Unconverted memb-Dendra2 (green), H2B-Cerulean (blue), photoconverted memb-Dendra2 (red) and a merged image of the three channels. (C) Photoconverted membranes are visible in the developing forebrain a day after photoconversion. Single focal plane images of the developing forebrain in a ventrally mounted zebrafish (anterior top) embryo 1 day post fertilization. The star indicates an individual photoconverted cell in mitosis during prometaphase. Left to Right: Unconverted memb-Dendra2 (green), H2B-Cerulean (blue), photoconverted memb-Dendra2 (red) and a merged image of the three channels. Scale Bar in (A) is ~150μm. Scale Bar in (B) and (C) is ~30μm.