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


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.


CONCLUSIONS
A tool is developed to isolate single cells to provide a suitable base for investigating and modelling the individual cell lag phase.
Francois
K
K
Laboratory of Food Microbiology and Preservation, Ghent University, Coupure Links 653, 9000 Ghent, Belgium.
Devlieghere
F
F
Standaert
A R
AR
Geeraerd
A H
AH
Van Impe
J F
JF
Debevere
J
J
D002455:Cell Division; D003470:Culture Media; D019649:Food Industry / Q000379:methods; D005516:Food Microbiology; D008089:Listeria monocytogenes / Q000166:cytology / Q000254:growth & development / Q000302:isolation & purification*; D008954:Models, Biological
D003470:Culture Media