Cargando…
High-throughput cultivation and screening platform for unicellular phototrophs
BACKGROUND: High-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. RESULTS: In this work we describe and...
Autores principales: | , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2014
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4172822/ https://www.ncbi.nlm.nih.gov/pubmed/25223876 http://dx.doi.org/10.1186/s12866-014-0239-x |
_version_ | 1782336077894254592 |
---|---|
author | Tillich, Ulrich M Wolter, Nick Schulze, Katja Kramer, Dan Brödel, Oliver Frohme, Marcus |
author_facet | Tillich, Ulrich M Wolter, Nick Schulze, Katja Kramer, Dan Brödel, Oliver Frohme, Marcus |
author_sort | Tillich, Ulrich M |
collection | PubMed |
description | BACKGROUND: High-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. RESULTS: In 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 CO(2) 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. CONCLUSIONS: The 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. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12866-014-0239-x) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-4172822 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-41728222014-10-23 High-throughput cultivation and screening platform for unicellular phototrophs Tillich, Ulrich M Wolter, Nick Schulze, Katja Kramer, Dan Brödel, Oliver Frohme, Marcus BMC Microbiol Methodology Article BACKGROUND: High-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. RESULTS: In 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 CO(2) 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. CONCLUSIONS: The 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. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12866-014-0239-x) contains supplementary material, which is available to authorized users. BioMed Central 2014-09-16 /pmc/articles/PMC4172822/ /pubmed/25223876 http://dx.doi.org/10.1186/s12866-014-0239-x Text en © Tillich et al.; licensee BioMed Central Ltd. 2014 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. |
spellingShingle | Methodology Article Tillich, Ulrich M Wolter, Nick Schulze, Katja Kramer, Dan Brödel, Oliver Frohme, Marcus High-throughput cultivation and screening platform for unicellular phototrophs |
title | High-throughput cultivation and screening platform for unicellular phototrophs |
title_full | High-throughput cultivation and screening platform for unicellular phototrophs |
title_fullStr | High-throughput cultivation and screening platform for unicellular phototrophs |
title_full_unstemmed | High-throughput cultivation and screening platform for unicellular phototrophs |
title_short | High-throughput cultivation and screening platform for unicellular phototrophs |
title_sort | high-throughput cultivation and screening platform for unicellular phototrophs |
topic | Methodology Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4172822/ https://www.ncbi.nlm.nih.gov/pubmed/25223876 http://dx.doi.org/10.1186/s12866-014-0239-x |
work_keys_str_mv | AT tillichulrichm highthroughputcultivationandscreeningplatformforunicellularphototrophs AT wolternick highthroughputcultivationandscreeningplatformforunicellularphototrophs AT schulzekatja highthroughputcultivationandscreeningplatformforunicellularphototrophs AT kramerdan highthroughputcultivationandscreeningplatformforunicellularphototrophs AT brodeloliver highthroughputcultivationandscreeningplatformforunicellularphototrophs AT frohmemarcus highthroughputcultivationandscreeningplatformforunicellularphototrophs |