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Updated: Aug 12, 2026
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The MultiBac Protein Complex Production Platform at the EMBL
Published on: July 11, 2013
Multimodal High-Throughput Screening Raman Spectroscopy for Label-Free Single-Cell Characterization of Recombinant
S M Miftahul Islam1, Kristina Worch2, Ayman Bali1
1Leibniz Institute of Photonic Technology , Albert-Einstein-Straße 9, Jena07745, Germany.
Analytical Chemistry
|August 11, 2026
Summary
This study introduces a new automated platform for analyzing recombinant protein expression in insect cells. It uses Raman spectroscopy, microscopy, and fluorescence to reveal single-cell biochemical and morphological differences, improving bioprocessing characterization.
Area of Science:
- Biotechnology
- Cell Biology
- Spectroscopy
Background:
- Recombinant protein expression using baculovirus expression vector systems (BEVS) faces challenges due to cell heterogeneity.
- Existing analytical methods often fail to simultaneously assess infection status, biochemical state, and cell morphology at the single-cell level.
Purpose of the Study:
- To develop and validate an automated multimodal platform for high-throughput, single-cell analysis of BEVS bioprocessing.
- To integrate Raman spectroscopy, phase-contrast microscopy, and fluorescence imaging for comprehensive cellular characterization.
- To investigate construct-dependent biochemical and morphological heterogeneity in insect cells.
Main Methods:
- An automated platform combining high-throughput Raman spectroscopy, phase-contrast microscopy, and fluorescence imaging was developed.
- Coregistered spectra and images were acquired from individual Spodoptera frugiperda (Sf) 9 cells.
- An integrated pipeline linked morphological, reporter abundance, and spectral data at single-cell resolution.
Main Results:
- A PCA-LDA model trained on Raman spectra achieved 93% cross-validation accuracy for classifying BEVS conditions.
- Raman spectroscopy provided biochemical insights distinct from fluorescence, capturing construct-dependent states.
- Phase-contrast microscopy revealed amplified cell swelling in dual-expression cells, indicating increased biosynthetic burden.
Conclusions:
- The multimodal platform enables simultaneous assessment of biochemical, morphological, and expression heterogeneity at single-cell resolution in BEVS.
- Correlated single-cell measurements offer significant analytical potential for characterizing and optimizing bioprocessing.
- This work establishes an analytical foundation for future at-line adaptation of BEVS monitoring.
