High-throughput cultivation and screening platform for unicellular phototrophs

BMC Microbiology

Methodology Article

Dan Kramer¹  Katja Schulze²  Marcus Frohme²  Oliver Brödel²  Nick Wolter²  Ulrich M Tillich³ 

[1] Cyano biotech GmbH, Magnusstrasse 11, D-12489, Berlin, Germany;Molecular Biotechnology and Functional Genomics, Technical University of Applied Sciences Wildau, Hochschulring 1, 16-2001, D-15745, Wildau, Germany;Molecular Biotechnology and Functional Genomics, Technical University of Applied Sciences Wildau, Hochschulring 1, 16-2001, D-15745, Wildau, Germany;Institute of Biology, Humboldt-University Berlin, Berlin, Germany;

关键词: Cyanobacteria;    Synechocystis;    HTS;    High throughput;    Automated cultivation;    Liquid handling;   

DOI  :  10.1186/s12866-014-0239-x

 received in 2014-05-09, accepted in 2014-08-27,  发布年份 2014

来源: Springer

【 摘 要 】

BackgroundHigh-throughput cultivation and screening methods allow a parallel, miniaturized and cost efficient processing of many samples. These methods however, have not been generally established for phototrophic organisms such as microalgae or cyanobacteria.ResultsIn this work we describe and test high-throughput methods with the model organism Synechocystis sp. PCC6803. The required technical automation for these processes was achieved with a Tecan Freedom Evo 200 pipetting robot. The cultivation was performed in 2.2 ml deepwell microtiter plates within a cultivation chamber outfitted with programmable shaking conditions, variable illumination, variable temperature, and an adjustable CO2 atmosphere. Each microtiter-well within the chamber functions as a separate cultivation vessel with reproducible conditions. The automated measurement of various parameters such as growth, full absorption spectrum, chlorophyll concentration, MALDI-TOF-MS, as well as a novel vitality measurement protocol, have already been established and can be monitored during cultivation. Measurement of growth parameters can be used as inputs for the system to allow for periodic automatic dilutions and therefore a semi-continuous cultivation of hundreds of cultures in parallel. The system also allows the automatic generation of mid and long term backups of cultures to repeat experiments or to retrieve strains of interest.ConclusionsThe presented platform allows for high-throughput cultivation and screening of Synechocystis sp. PCC6803. The platform should be usable for many phototrophic microorganisms as is, and be adaptable for even more. A variety of analyses are already established and the platform is easily expandable both in quality, i.e. with further parameters to screen for additional targets and in quantity, i.e. size or number of processed samples.

【 授权许可】

Unknown   
© Tillich et al.; licensee BioMed Central Ltd. 2014. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.