Microscopy Imaging of Fluorescent Cells

Image acquisition and processing was performed as described previously61 (link). Cells were analysed with a microscope (Axiovert 200 M; Carl Zeiss MicroImaging, Inc.) equipped with an Exfo X-cite 120 excitation light source, band pass filters (Carl Zeiss MicroImaging, Inc. and Chroma Technology Corp.), an α Plan-Fluar 100x/1.45 NA, Plan-Apochromat 63x/1.4 NA or A-plan (Carl Zeiss MicroImaging, Inc.), and a digital camera (Orca ER; Hamamatsu). Image acquisition was performed using Volocity software (Improvision). Fluorescence images were collected as 0.5 μm Z-stacks using exposures of up to 200 ms, merged into one plane in Openlab, and processed further in Photoshop (Adobe). Brightfield images were collected in one plane, and processed where necessary to highlight circumference of the cells.