Publication Details

Category Text Publication
Reference Category Journals
DOI 10.1093/pnasnexus/pgad006
Licence creative commons licence
Title (Primary) Live tracking metabolic networks and physiological responses within microbial assemblages at single-cell level
Author Cui, L.; Xin, Y.; Yang, K.; Li, H.; Tan, F.; Zhang, Y.; Li, X.; Zhu, Z.; Yang, J.; Kao, S.-J.; Ren, B.; Zhu, Y.-G.; Musat, F.; Musat, N.
Source Titel PNAS Nexus
Year 2023
Department ISOBIO
Volume 2
Issue 3
Page From pgad006
Language englisch
Topic T7 Bioeconomy
Supplements https://oup.silverchair-cdn.com/oup/backfile/Content_public/Journal/pnasnexus/2/3/10.1093_pnasnexus_pgad006/1/pgad006_supplementary_data.pdf?Expires=1681986709&Signature=WmtEUr~PsmeQ9qfNmHmSAXiCPn8TF8B6S3gnzafXblLkxBusbvjV8aijk7Ph9p54-K7NQyRqIlJl0iJWzAQ2tsgXOZ97pdYJOiXxH2ixaz27fbnO5TS~KrMPlG8UMXPqYw0KUvwaNksYMSUm0iuTWQGT97KkeI5GS5J6Z8iYggYESshso1RumxAkdT98~1bqLa6Z~4FMUXeEK6ryD6h-aJlKAX2kpkp3ojYV~oVlPxlOkA-uL~NZFyWmzr2PpxMOVoI--ux-~o3EirGml2SlkWjNDMMoHYZEdH7Mm20466jCMz3ErrctFdhV5cb9jqV84voVTNSeGR2PbzWPHj9c2Q__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA
Keywords microbial interactions; single-cell Raman spectroscopy; stable isotope probing; nitrogen fixation; carbon fixation
Abstract Microbial interactions impact the functioning of both natural and engineered systems, yet our ability to directly monitor these highly dynamic and spatially resolved interactions in living cells is very limited. Here, we developed a synergistic approach coupling single-cell Raman microspectroscopy with 15N2 and 13CO2 stable isotope probing in a microfluidic culture system (RMCS-SIP) for live tracking of the occurrence, rate, and physiological shift of metabolic interactions in active microbial assemblages. Quantitative and robust Raman biomarkers specific for N2 and CO2 fixation in both model and bloom-forming diazotrophic cyanobacteria were established and cross-validated. By designing a prototype microfluidic chip allowing simultaneous microbial cultivation and single-cell Raman acquisition, we achieved temporal tracking of both intercellular (between heterocyst and vegetative cells of cyanobacteria) and interspecies N and C metabolite exchange (from diazotroph to heterotroph). Moreover, single-cell N and C fixation and bidirectional transfer rate in living cells were quantified via SIP-induced characteristic Raman shifts. Remarkably, RMCS captured physiological responses of metabolically active cells to nutrient stimuli through comprehensive metabolic profiling, providing multimodal information on the evolution of microbial interactions and functions under fluctuating conditions. This noninvasive RMCS-SIP is an advantageous approach for live-cell imaging and represents an important advancement in the single-cell microbiology field. This platform can be extended for real-time tracking of a wide range of microbial interactions with single-cell resolution and advances the understanding and manipulation of microbial interactions for societal benefit.
Persistent UFZ Identifier https://www.ufz.de/index.php?en=20939&ufzPublicationIdentifier=23865
Cui, L., Xin, Y., Yang, K., Li, H., Tan, F., Zhang, Y., Li, X., Zhu, Z., Yang, J., Kao, S.-J., Ren, B., Zhu, Y.-G., Musat, F., Musat, N. (2023):
Live tracking metabolic networks and physiological responses within microbial assemblages at single-cell level
PNAS Nexus 2 (3), pgad006 10.1093/pnasnexus/pgad006