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@ARTICLE{Jansen:890775,
author = {Jansen, Roman and Küsters, Kira and Morschett, Holger and
Wiechert, Wolfgang and Oldiges, Marco},
title = {{A} fully automated pipeline for the dynamic at-line
morphology analysis of microscale {A}spergillus cultivation},
journal = {Fungal Biology and Biotechnology},
volume = {8},
number = {2},
issn = {2054-3085},
address = {London},
publisher = {BioMed Central},
reportid = {FZJ-2021-01191},
pages = {1-10},
year = {2021},
abstract = {BackgroundMorphology, being one of the key factors
influencing productivity of filamentous fungi, is of great
interest during bioprocess development. With increasing
demand of high-throughput phenotyping technologies for fungi
due to the emergence of novel time-efficient genetic
engineering technologies, workflows for automated liquid
handling combined with high-throughput morphology analysis
have to be developed.ResultsIn this study, a protocol
allowing for 48 parallel microbioreactor cultivations of
Aspergillus carbonarius with non-invasive online signals of
backscatter and dissolved oxygen was established. To handle
the increased cultivation throughput, the utilized
microbioreactor is integrated into a liquid handling
platform. During cultivation of filamentous fungi, cell
suspensions result in either viscous broths or form pellets
with varying size throughout the process. Therefore,
tailor-made liquid handling parameters such as
aspiration/dispense height, velocity and mixing steps were
optimized and validated. Development and utilization of a
novel injection station enabled a workflow, where biomass
samples are automatically transferred into a flow through
chamber fixed under a light microscope. In combination with
an automated image analysis concept, this enabled an
automated morphology analysis pipeline. The workflow was
tested in a first application study, where the projected
biomass area was determined at two different cultivation
temperatures and compared to the microbioreactor online
signals.ConclusionsA novel and robust workflow starting from
microbioreactor cultivation, automated sample harvest and
processing via liquid handling robots up to automated
morphology analysis was developed. This protocol enables the
determination of projected biomass areas for filamentous
fungi in an automated and high-throughput manner. This
measurement of morphology can be applied to describe overall
pellet size distribution and heterogeneity.},
cin = {IBG-1},
ddc = {630},
cid = {I:(DE-Juel1)IBG-1-20101118},
pnm = {2172 - Utilization of renewable carbon and energy sources
and engineering of ecosystem functions (POF4-217)},
pid = {G:(DE-HGF)POF4-2172},
typ = {PUB:(DE-HGF)16},
pubmed = {33676585},
doi = {10.1186/s40694-021-00109-4},
url = {https://juser.fz-juelich.de/record/890775},
}