Drummond:Fluorescence: Difference between revisions
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#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) | #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) | ||
#normalize OD to 0.4 | #normalize OD to 0.4 | ||
target absorbance (0.4)/original absorbance = ratio | |||
4000 μL (4 mL) * (ratio) = X μL preculture in 4 mL solution | 4000 μL (4 mL) * (ratio) = X μL preculture in 4 mL solution | ||
==Materials== | ==Materials== | ||
Revision as of 12:41, 20 June 2008
Introduction
Goal: to prepare yeast cells with RFP/TFP fluorescence for imaging on the LSM510.
Protocol
- day before imaging: prepare yeast precultures in 2 mL YPsugar (YPD, YPsuc/raf)
- 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)
- normalize OD to 0.4
target absorbance (0.4)/original absorbance = ratio 4000 μL (4 mL) * (ratio) = X μL preculture in 4 mL solution
