Three step flow focusing enables image-based discrimination and sorting of late stage 1 Haematococcus pluvialis cells

Daniel Kraus; Andreas Kleiber; Enrico Ehrhardt; Matthias Leifheit; Peter Horbert; Matthias Urban; Nils Gleichmann; Günter Mayer; Jürgen Popp; Thomas Henkel

doi:10.1371/journal.pone.0249192

Three step flow focusing enables image-based discrimination and sorting of late stage 1 Haematococcus pluvialis cells

PLoS One. 2021 Mar 29;16(3):e0249192. doi: 10.1371/journal.pone.0249192. eCollection 2021.

Authors

Affiliations

¹ Leibniz Institute of Photonic Technology, Jena, Germany.
² Gesellschaft zur Förderung von Medizin-, Bio- und Umwelttechnologien e. V. (GMBU), Halle (Saale), Germany.
³ Institute of Physical Chemistry and Abbe Center of Photonics, Friedrich-Schiller-University, Jena, Germany.

Abstract

Label-free and gentle separation of cell stages with desired target properties from mixed stage populations are a major research task in modern biotechnological cultivation process and optimization of micro algae. The reported microfluidic sorter system (MSS) allows the subsequent investigation of separated subpopulations. The implementation of a viability preserving MSS is shown for separation of late stage 1 Haematococcus pluvialis (HP) cells form a mixed stage population. The MSS combines a three-step flow focusing unit for aligning the cells in single file transportation mode at the center of the microfluidic channel with a pure hydrodynamic sorter structure for cell sorting. Lateral displacement of the cells into one of the two outlet channels is generated by piezo-actuated pump chambers. In-line decision making for sorting is based on a user-definable set of image features and properties. The reported MSS significantly increased the purity of target cells in the sorted population (94%) in comparison to the initial mixed stage population (19%).

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Separation / instrumentation*
Chlorophyceae / cytology*
Lab-On-A-Chip Devices*

Grants and funding

This study is part of the collaborative research center AquaDiva (CRC 1076 AquaDiva) funded by the Deutsche Forschungsgemeinschaft (DFG). The funding from Bundesministerium für Wirtschaft und Technologie to AK (Pollen3D–ZF4006816CR8) and Thüringer Aufbaubank to DK (2018 SD 0066) is gratefully acknowledged. The publication of this article was funded by the Open Access Fund of the Leibniz Association. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.