Figure 3

Photoconversion of widespread expressing CAG::KikGR transgenic ES cells. Panels show 2D (A-F, I) and 3D images (G, H) of the photoconversion of a selected group of cells in an ES cell colony. Bright field image of ES cell colony (A). Image of ES cell colony with green (B) and red (C) channel showing green but no red fluorescence prior to photoconversion. Yellow dashed box show regions of interest (ROI) that is to be photoconverted. After photoconversion, cells in an ROI do not fluoresce green (D) but red (E). Merge of green and red channel after photoconversion, showing complete photoconversion of cells in the region of interest. 3D image of ES cell colony before photoconversion. All cells emit green fluorescence (G). Line plot showing average fluorescent intensity measured along the white arrow in G (G'). 3D image of ES cell colony after photoconversion. Photoconverted cells emit red fluorescence (H). Line plot showing average fluorescent intensity measured along the white arrow in H showing complete photoconversion of cells, i.e. shift of emission spectrum from green to red in photoconverted region (H'). 2D orthogonal representation of photoconverted ES cell colony showing complete photoconversion in the region of interest through entire z-stack (I).