Home › Microbiology › ODELAY: A Large-scale Method for Multi-parameter Quantification of Yeast Growth
Microbiology JoVE (Open Access) Citable · DOI

ODELAY: A Large-scale Method for Multi-parameter Quantification of Yeast Growth

DOI: 10.3791/55879-v
What you'll learn
  • ✓ Assemble agar molds and prepare yeast cultures for ODELAY analysis
  • ✓ Perform automated liquid spotting and time-lapse microscopy setup
  • ✓ Quantify single-cell growth heterogeneity across yeast colony development phases
Protocol

We present a method for quantifying growth phenotypes of individual yeast cells as they grow into colonies on solid media using time-lapse microscopy termed, One-cell Doubling Evaluation of Living Arrays of Yeast (ODELAY). Population heterogeneity of genetically identical cells growing into colonies can be directly observed and quantified.

Difficulty
advanced
Total time
~3–5 days (includes overnight culture preparation, imaging, and post-acquisition analysis)
Model organism
Saccharomyces cerevisiae (budding yeast)
Biosafety
BSL-1

Steps

1
Assemble agar molds for colony growth

Prepare and assemble agar molds that will serve as the solid media substrate for yeast colony growth during the ODELAY experiment.

▶ 00:39
2
Prepare yeast cultures for spotting

Culture and prepare genetically identical yeast cells at appropriate density for subsequent spotting onto agar plates.

▶ 03:36
3
Spot yeast cultures using automated robot

Use an automated liquid spotting robot to deposit precise, uniform volumes of yeast culture onto agar molds to initiate colony growth.

▶ 05:41
4
Acquire time-lapse microscopy images

Set up and execute automated time-lapse microscopy imaging to capture yeast colony development and individual cell growth dynamics over time.

▶ 07:21
5
Analyze growth phases and population heterogeneity

Process time-lapse microscopy data to quantify and characterize all three growth phases of yeast cells and measure phenotypic heterogeneity within genetically identical populations.

▶ 09:03
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