Bacterial Isolate Microscopy

Epifluorescence microscopy of gut bacterial isolates after EdU-click. Human gut bacterial isolates which underwent EdU-click with or without EdU were stained with Sybr Green I nucleic acid dye and visualized using epifluorescence microscopy. Green = Sybr Green I, staining all bacterial nucleic acid content; Red = Alexa Fluor 647 azide, staining only newly replicated bacterial DNA.

All slides were visualized using the Axiovert 3 widefield inverted microscope (Zeiss) equipped with a Zeiss Axiocam 506 monochrome CCD camera (2752 x 2208 pixels, 4.54 um pixels) and Sola SM solid state light engine, using the 100X objective (PLAN-APOCHROMAT, NA=1.4, OIL, Iris) under oil immersion and the FS 24 (FITC) and FS 32 (Cy5) fluorescence cubes. Images were further processed in Fiji (version 1.53q)