During image capture, 5–7 images were taken on each spot. The exposure time was set so that PLA signals were easily distinguishable but not overexposed. It varied between 200 and 500 ms for TRITC with 10% exposure and 200–400 ms for DAPI with 5% exposure. Each image was captured on one layer in the focus plane (DAPI). For TRITC filter, few signals can be visualized either above or below the current focus but the low thickness of the cells after the cytospin allowed all dots to be visible in one focus. If necessary, to increase the resolution, 8 layers (separated for instance by 0.3 nm each) can be taken and a "Z-projection" on the maximum intensity can be applied on the picture. The captured images can be saved as .dv or .tiff.