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J Vis Exp. 2014 Jul 15;(89). doi: 10.3791/51456.

ScanLag: high-throughput quantification of colony growth and lag time.

Author information

1
Racah Institute of Physics, The Hebrew University of Jerusalem.
2
Racah Institute of Physics, The Hebrew University of Jerusalem; nathalieqb@phys.huji.ac.il.

Abstract

Growth dynamics are fundamental characteristics of microorganisms. Quantifying growth precisely is an important goal in microbiology. Growth dynamics are affected both by the doubling time of the microorganism and by any delay in growth upon transfer from one condition to another, the lag. The ScanLag method enables the characterization of these two independent properties at the level of colonies originating each from a single cell, generating a two-dimensional distribution of the lag time and of the growth time. In ScanLag, measurement of the time it takes for colonies on conventional nutrient agar plates to be detected is automated on an array of commercial scanners controlled by an in house application. Petri dishes are placed on the scanners, and the application acquires images periodically. Automated analysis of colony growth is then done by an application that returns the appearance time and growth rate of each colony. Other parameters, such as the shape, texture and color of the colony, can be extracted for multidimensional mapping of sub-populations of cells. Finally, the method enables the retrieval of rare variants with specific growth phenotypes for further characterization. The technique could be applied in bacteriology for the identification of long lag that can cause persistence to antibiotics, as well as a general low cost technique for phenotypic screens.

PMID:
25077667
PMCID:
PMC4215631
DOI:
10.3791/51456
[Indexed for MEDLINE]
Free PMC Article

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