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Lett Appl Microbiol. 2003;37(1):26-30.

Modelling the individual cell lag phase. Isolating single cells: protocol development.

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1
Laboratory of Food Microbiology and Preservation, Ghent University, Coupure Links 653, 9000 Ghent, Belgium.

Abstract

AIMS:

To develop a protocol to isolate single cells in wells of a microtitre plate, having a high certainty of individual cells, combined with a sufficient yield.

METHODS AND RESULTS:

Single cells were obtained using 1/2 dilution series in microtitre plates. Seventy-two Lactococcus lactis dilution series were checked by plate counting. When the last five columns of the plates were observed, the chance of having one single cell was 80%, while the yield was 75 wells containing cells. A simulation model confirmed these results. This method was compared with the commonly applied method.

CONCLUSIONS:

This method makes it possible to combine a higher chance of having one cell in a microtitre well with a slightly higher yield.

SIGNIFICANCE AND IMPACT OF THE STUDY:

A tool is developed to isolate single cells to provide a suitable base for investigating and modelling the individual cell lag phase.

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