Longitudinal Single-Cell Imaging of Engineered Strains with Stimulated Raman Scattering to Characterize Heterogeneity in Fatty Acid Production.

Autor: Tague N; Department of Biomedical Engineering, Boston University, Boston, MA, 02215, USA.; Biological Design Center, Boston University, Boston, MA, 02215, USA., Lin H; Department of Biomedical Engineering, Boston University, Boston, MA, 02215, USA.; Photonics Center, Boston University, Boston, MA, 02215, USA., Lugagne JB; Department of Biomedical Engineering, Boston University, Boston, MA, 02215, USA.; Biological Design Center, Boston University, Boston, MA, 02215, USA., O'Connor OM; Department of Biomedical Engineering, Boston University, Boston, MA, 02215, USA.; Biological Design Center, Boston University, Boston, MA, 02215, USA., Burman D; Department of Biomedical Engineering, Boston University, Boston, MA, 02215, USA., Wong WW; Department of Biomedical Engineering, Boston University, Boston, MA, 02215, USA.; Biological Design Center, Boston University, Boston, MA, 02215, USA., Cheng JX; Department of Biomedical Engineering, Boston University, Boston, MA, 02215, USA.; Photonics Center, Boston University, Boston, MA, 02215, USA.; Department of Electrical and Computer Engineering, Boston University, Boston, MA, 02215, USA., Dunlop MJ; Department of Biomedical Engineering, Boston University, Boston, MA, 02215, USA.; Biological Design Center, Boston University, Boston, MA, 02215, USA.
Jazyk: angličtina
Zdroj: Advanced science (Weinheim, Baden-Wurttemberg, Germany) [Adv Sci (Weinh)] 2023 Jul; Vol. 10 (20), pp. e2206519. Date of Electronic Publication: 2023 Jun 08.
DOI: 10.1002/advs.202206519
Abstrakt: Understanding metabolic heterogeneity is critical for optimizing microbial production of valuable chemicals, but requires tools that can quantify metabolites at the single-cell level over time. Here, longitudinal hyperspectral stimulated Raman scattering (SRS) chemical imaging is developed to directly visualize free fatty acids in engineered Escherichia coli over many cell cycles. Compositional analysis is also developed to estimate the chain length and unsaturation of the fatty acids in living cells. This method reveals substantial heterogeneity in fatty acid production among and within colonies that emerges over the course of many generations. Interestingly, the strains display distinct types of production heterogeneity in an enzyme-dependent manner. By pairing time-lapse and SRS imaging, the relationship between growth and production at the single-cell level are examined. The results demonstrate that cell-to-cell production heterogeneity is pervasive and provides a means to link single-cell and population-level production.
(© 2023 The Authors. Advanced Science published by Wiley-VCH GmbH.)
Databáze: MEDLINE
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