Generation of larger numbers of separated microbial populations by cultivation in segmented-flow microdevices.
The high speed production of fluid segments for the highly parallelized cultivation of monoclonal cell populations was carried out by the use of microchip segmentor modules. Aqueous fluid segments, embedded in a non-miscible carrier liquid, were produced with frequencies up to 30 s(-1) and showed a high homogeneity in size. This corresponds with the production of about 2.5 million samples per day. The segment volumes can be adapted between about 4 nl and 100 nl. The typical segment size for cultivation experiments is in the range between 40 nl and 80 nl. Nutrient medium can be applied instead of pure water. It is possible to aliquot a cell suspension in such a way that most of the aqueous fluid segments contain only one cell. In model experiments with four microbial species chip-produced aliquots of 60 nl, each containing one or a few cells, were incubated in Teflon capillary tubes. Rapid growth of the microcultures was observed. Cell densities were found to be as high as in conventional shake flask cultures.
Martin
Karin
K
Hans Knöll Institute for Natural Products Research, Jena, Germany.
Henkel
Thomas
T
Baier
Volker
V
Grodrian
Andreas
A
Schön
Thore
T
Roth
Martin
M
Michael Köhler
Johann
J
Metze
Josef
J
D015169:Colony Count, Microbial; D003470:Culture Media; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development; D005453:Fluorescence; D008828:Microbiological Techniques / Q000295:instrumentation* / Q000379:methods*; D044085:Microfluidics / Q000295:instrumentation*; D011138:Polytetrafluoroethylene; D011551:Pseudomonas fluorescens / Q000235:genetics / Q000254:growth & development; D012240:Rhodococcus / Q000235:genetics / Q000254:growth & development; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development; D013997:Time Factors
D003470:Culture Media; D011138:Polytetrafluoroethylene