G box f3 gel imager

Exponentially growing wild-type cells were either washed with sulfate-free medium for sulfur starvation or subjected to different concentrations of sodium sulfide in SD medium. met17Δ were grown to exponential phase in methionine-supplemented SD and washed with SD without organosulfur supplements. In all treatments, cultures were set up at an OD of approximately 0.1, volume 2.5 ml, in 13-mm glass tubes. Tubes were sealed with parafilm for sulfide treatment. For imaging at different time points, 4 μl of the cell culture was spread under a coverslip on a slide. The fluorescence microscopy setup comprised a Nikon Eclipse TE-2000U inverted fluorescence microscope (Nikon, Tokyo, Japan) connected to a cooled CCD camera for fluorescence and transmitted light imaging. Image acquisition was done with an in-house LabVIEW program. Images were captured using either a 100× objective (Fig 4A–4D) or a 40× objective (Fig 5B and Fig Qi in S1 Figs). For imaging GFP, the filter cube used was Chroma 49002-ET-EGFP (exciter: 470/40×, emitter: 525/50m, Dichroic: T495LP; Chroma Technology, Bellows Falls, VT, USA). Imaging conditions and exposure times were kept constant for imaging different treatments. Images of yeast growth on agar plates were captured using a G:BOX F3 gel imager (Syngene, Cambridge, UK). Image processing was done on Fiji [61 (link)].