Abstract
In the development and optimization of biotechnological cultivation processes the continuous monitoring through the acquisition and interpretation of spectral and morphological properties of bioparticles are challenging. There is therefore a need for the parallel acquisition and interpretation of spatially and spectrally resolved measurements with which particles can be characterized and classified in-flow with high throughput. Therefore, in this paper we investigated the scientific and technological connectivity of standard imaging flow cytometry (IFC) with filter-on-chip based spatially and spectrally resolving snapshot-mosaic cameras for photonic sensing and control in a smart and innovative microfluidic device. For the investigations presented here we used the microalgae Haematococcus pluvialis (HP). These microalgae are used commercially to produce the antioxidant keto-carotenoid astaxanthin. Therefore, HP is relevant to practically demonstrate the usability of the developed system for Multispectral Imaging Flow Cytometry (MIFC) platform. The extension of standard IFC with snapshot-mosaic cameras and multivariate data processing is an innovative approach for the in-flow characterization and derived classification of bioparticles. Finally, the multispectral data acquisition and the therefore developed methodology is generalizable and enables further applications far beyond the here characterized population of HP cells.
Highlights
For the multivariate data processing and spectral model building we use the software fluxTrainer from LuxFlux GmbH (Reutlingen, Germany). This software allows complex multivariate data analysis and processing with spectral model building. With this software we developed a spectral model for the classification of Haematococcus pluvialis (HP) cells on real multispectral data and perform data interpretation and result writing as complex .h5 files
To acquire and process data from snapshot-mosaic cameras we use .xml files with first- and second order software we developed a spectral model for the classification of HP cells on real multispectral data and perform data interpretation and result writing as complex .h5 files
This is necessary because the behavior of the used snapshotmosaic camera camera concerning concerning the the spectral spectral sensitivity sensitivity differs differs from from the the generalized generalized provided provided mosaic manufacturers data
Summary
In the development and optimization of biotechnological cultivation processes the continuous monitoring through the acquisition and interpretation of spectral and morphological properties of bioparticles are challenging. There is a need for the parallel acquisition and interpretation of spatially and spectrally resolved measurements with which particles can be characterized and classified in-flow with high throughput. In this paper we investigated the scientific and technological connectivity of standard imaging flow cytometry (IFC) with filter-on-chip based spatially and spectrally resolving snapshot-mosaic cameras for photonic sensing and control in a smart and innovative microfluidic device. The extension of standard IFC with snapshot-mosaic cameras and multivariate data processing is an innovative approach for the in-flow characterization and derived classification of bioparticles. Multispectral Imaging Flow Cytometry (MIFC) combines the spectral with spatially resolved morphological properties of particles.
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