Figure S1.

Microscope and µManager plugin for photoactivation experiments. (a) A DMD and a blue LED (centered around 405 nm) light source were engineered on a Nikon Eclipse Ti-E microscope as shown in the figure. A computer was used to control the DMD, which reflects light into the microscope only when pixels are in the “on” position, so displaying a mask matching the cell photoactivates that cell. (b) Example images of a photoactivation experiment. Cells (hTERT-RPE1 PA-mCherry H2B-mGFP) are shown imaged in the GFP channel (green), during photoactivation (blue light channel, blue), and before and after photoactivation (mCherry channel, red). Scale bar: 100 µm. (c) A μManager plugin was developed to enable automatic image acquisition, analysis, and photoactivation. An analysis plugin defines its own set of parameters that can be manipulated by the user. Two analysis plugins were used in this study, one for cell identification and another for nuclear size measurement.