Drummond:Fluorescence

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Introduction

Goal: to prepare yeast cells with RFP/TFP fluorescence for imaging on the LSM510.

Protocol

  1. day before imaging: prepare yeast precultures in 2 mL YPsugar (YPD, YPsuc/raf)
  2. day of imaging (allow for >3 hour induction): measure optical density (OD) of preculture. Dilution may be necessary (1:20 dilution of preculture:growth media; 50 μL preculture/950 μL YPsugar)
  3. normalize OD to 0.4

target absorbance (0.4)/original absorbance = ratio 4000 μL (4 mL) * (ratio) = X μL preculture in 4 mL solution

Materials

References

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