Yeast strains harboring a GFP-tag were grown at 25°C to early log phase (OD600 0.4–0.6) in selective SC medium. Fluorescence imaging was performed as described previously (Liu and Novick, 2014 (link)). In brief, images were acquired with a 100× oil immersion objective lens (Plan Apochromat 100×/1.4 NA oil DIC lens; Carl Zeiss) on a spinning disk confocal microscopy system (Yokogawa Electric Corporation), which includes a microscope (Observer Z1; Carl Zeiss) equipped with an electron multiplying charge-coupled device camera (QuantEM 512SC; Photometrics). Excitation of GFP or mCherry was achieved using 488-nm argon and 568-nm argon/krypton lasers, respectively. Images were analyzed using AxioVision software 4.8 (Carl Zeiss).
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Fluorescence Imaging of GFP-Tagged Yeast
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Corresponding Organization : University of California, San Diego
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Variable analysis
independent variables
- Growth temperature (25°C)
- Fluorescence intensity (GFP and mCherry)
- Growth phase (early log phase, OD600 0.4-0.6)
- Growth medium (selective SC medium)
- Positive control: None specified
- Negative control: None specified
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