Combining coherent Raman scattering with fluorescence microscopy to investigate sugar uptake in living cells

• Applicant: Dr. Evelyn Plötz
• Subject Area: Biophysics ; Analytical Chemistry
• Term: from 2019 to 2024
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 426814631

• Final Report Year: 2024

Final Report Abstract

Understanding the roles of sugars in cellular processes is crucial due to their importance in metabolic pathways and glycosylation processes. Glycans, built up from single monosaccharides, impact processes like cell signaling, adhesion, and form the glycocalyx. Changes in glycosylation often relate to pathological conditions, such as inflammation and tumorigenesis, highlighting the need to understand sugar composition and function. While structural approaches provide detailed glycome analysis, they lack spatial information about glycan distribution within cells. Optical microscopy, especially fluorescence imaging, offers insights into subcellular structures and dynamic processes. Metabolic oligosaccharide engineering allows for sugar labeling in cells. Yet, the incorporation path of unnatural sugars into cells remains unclear, necessitating innovative imaging approaches. We aimed to implement stimulated Raman scattering (SRS) microscopy, combined with fluorescence-based live-cell imaging, to study GLUT transporter mediated sugar uptake and storage in cells. Good progress was made in developing an SRS microscope, combining it with confocal fluorescence microscopy, and developing acquisition software for correlative, multimodal microscopy. The established MOSAIC microscopy setup allows for correlative multi-color fluorescence imaging, nonlinear microscopy and space-resolved Raman spectroscopy for in-situ measurements. We benchmarked the imaging system for transparent, solid state samples like graphene, and optimized it for soft-matter imaging through projects involving tissue sections, spheroids and single cells, that helped us to expand MOSAIC even to Fluorescence-Lifetime microscopy. As cellular sugar uptake is tightly regulated, we lastly explored metal-organic-framework (MOF) as carriers for sugars and studied various MOFs at the single-particle level. Our research uncovered new material properties and their significant impact on parameters, such as optical response or uptake capacities for guest-molecules. We used MOSAIC to investigate ‘small’ molecules in MOFs employed for atmospheric water harvesting, hydrogen storage and CO2 capture. Moving forward, we currently develop advanced microscopy tools to track MOF nanoparticle dynamics in within cellular environments.

Publications

• Metal–Organic Framework Nanoparticles Induce Pyroptosis in Cells Controlled by the Extracellular pH. Advanced Materials, 32(19).
  Ploetz, Evelyn; Zimpel, Andreas; Cauda, Valentina; Bauer, David; Lamb, Don C.; Haisch, Christoph; Zahler, Stefan; Vollmar, Angelika M.; Wuttke, Stefan & Engelke, Hanna
  
• Modular Assembly of Red Blood Cell Superstructures from Metal–Organic Framework Nanoparticle‐Based Building Blocks. Advanced Functional Materials, 31(10).
  Guo, Jimin; Yu, Yunlong; Zhu, Wei; Serda, Rita E.; Franco, Stefan; Wang, Lu; Lei, Qi; Agola, Jacob Ongudi; Noureddine, Achraf; Ploetz, Evelyn; Wuttke, Stefan & Brinker, C. Jeffrey
  
• The Chemistry of Reticular Framework Nanoparticles: MOF, ZIF, and COF Materials. Advanced Functional Materials, 30(41).
  Ploetz, Evelyn; Engelke, Hanna; Lächelt, Ulrich & Wuttke, Stefan
  
• Applications of reticular diversity in metal–organic frameworks: An ever-evolving state of the art. Coordination Chemistry Reviews, 430, 213655.
  Ejsmont, Aleksander; Andreo, Jacopo; Lanza, Arianna; Galarda, Aleksandra; Macreadie, Lauren; Wuttke, Stefan; Canossa, Stefano; Ploetz, Evelyn & Goscianska, Joanna
  
• Artificial Bioaugmentation of Biomacromolecules and Living Organisms for Biomedical Applications. ACS Nano, 15(3), 3900-3926.
  Cao, Jiangfan; Zaremba, Orysia T.; Lei, Qi; Ploetz, Evelyn; Wuttke, Stefan & Zhu, Wei
  
• FRET-based dynamic structural biology: Challenges, perspectives and an appeal for open-science practices. eLife, 10.
  Lerner, Eitan; Barth, Anders; Hendrix, Jelle; Ambrose, Benjamin; Birkedal, Victoria; Blanchard, Scott C.; Börner, Richard; Sung, Chung Hoi; Cordes, Thorben; Craggs, Timothy D.; Deniz, Ashok A.; Diao, Jiajie; Fei, Jingyi; Gonzalez, Ruben L.; Gopich, Irina V.; Ha, Taekjip; Hanke, Christian A.; Haran, Gilad; Hatzakis, Nikos S. ... & Weiss, Shimon
  
• Graphene-on-Glass Preparation and Cleaning Methods Characterized by Single-Molecule DNA Origami Fluorescent Probes and Raman Spectroscopy. ACS Nano, 15(4), 6430-6438.
  Krause, Stefan; Ploetz, Evelyn; Bohlen, Johann; Schüler, Patrick; Yaadav, Renukka; Selbach, Florian; Steiner, Florian; Kamińska, Izabela & Tinnefeld, Philip
  
• Single Crystals Heterogeneity Impacts the Intrinsic and Extrinsic Properties of Metal–Organic Frameworks. Advanced Materials, 34(3).
  Fuchs, Adrian; Mannhardt, Petra; Hirschle, Patrick; Wang, Haoze; Zaytseva, Irina; Ji, Zhe; Yaghi, Omar; Wuttke, Stefan & Ploetz, Evelyn
  
• Spatio-selective activation of nuclear translocation of YAP with light directs invasion of cancer cell spheroids. iScience, 24(3), 102185.
  Illes, Bernhard; Fuchs, Adrian; Gegenfurtner, Florian; Ploetz, Evelyn; Zahler, Stefan; Vollmar, Angelika M. & Engelke, Hanna
  
• Sponges as bioindicators for microparticulate pollutants?. Environmental Pollution, 268, 115851.
  Girard, Elsa B.; Fuchs, Adrian; Kaliwoda, Melanie; Lasut, Markus; Ploetz, Evelyn; Schmahl, Wolfgang W. & Wörheide, Gert
  
• Structural and biophysical characterization of the tandem substrate-binding domains of the ABC importer GlnPQ. Open Biology, 11(4).
  Ploetz, Evelyn; Schuurman-Wolters, Gea K.; Zijlstra, Niels; Jager, Amarins W.; Griffith, Douglas A.; Guskov, Albert; Gouridis, Giorgos; Poolman, Bert & Cordes, Thorben
  
• The Current Status of MOF and COF Applications. Angewandte Chemie International Edition, 60(45), 23975-24001.
  Freund, Ralph; Zaremba, Orysia; Arnauts, Giel; Ameloot, Rob; Skorupskii, Grigorii; Dincă, Mircea; Bavykina, Anastasiya; Gascon, Jorge; Ejsmont, Aleksander; Goscianska, Joanna; Kalmutzki, Markus; Lächelt, Ulrich; Ploetz, Evelyn; Diercks, Christian S. & Wuttke, Stefan
  
• Chemical Synthesis of the Fluorescent, Cyclic Dinucleotides cthGAMP. ChemBioChem, 23(8).
  Veth, Simon; Fuchs, Adrian; Özdemir, Dilara; Dialer, Clemens; Drexler, David Jan; Knechtel, Fabian; Witte, Gregor; Hopfner, Karl‐Peter; Carell, Thomas & Ploetz, Evelyn
  
• Reticular Nanoscience: Bottom-Up Assembly Nanotechnology. Journal of the American Chemical Society, 144(17), 7531-7550.
  Andreo, Jacopo; Ettlinger, Romy; Zaremba, Orysia; Peña, Quim; Lächelt, Ulrich; de Luis, Roberto Fernández; Freund, Ralph; Canossa, Stefano; Ploetz, Evelyn; Zhu, Wei; Diercks, Christian S.; Gröger, Harald & Wuttke, Stefan
  
• System of sequences in multivariate reticular structures. Nature Reviews Materials, 8(5), 331-340.
  Canossa, Stefano; Ji, Zhe; Gropp, Cornelius; Rong, Zichao; Ploetz, Evelyn; Wuttke, Stefan & Yaghi, Omar M.
  
• A new twist on PIFE: photoisomerisation-related fluorescence enhancement. Methods and Applications in Fluorescence, 12(1), 012001.
  Ploetz, Evelyn; Ambrose, Benjamin; Barth, Anders; Börner, Richard; Erichson, Felix; Kapanidis, Achillefs N.; Kim, Harold D.; Levitus, Marcia; Lohman, Timothy M.; Mazumder, Abhishek; Rueda, David S.; Steffen, Fabio D.; Cordes, Thorben; Magennis, Steven W. & Lerner, Eitan
  
• Conquering Metal–Organic Frameworks by Raman Scattering Techniques. Advanced Functional Materials, 34(43).
  Tittel, Jonas; Knechtel, Fabian & Ploetz, Evelyn
  
• Deep-LASI: deep-learning assisted, single-molecule imaging analysis of multi-color DNA origami structures. Nature Communications, 14(1).
  Wanninger, Simon; Asadiatouei, Pooyeh; Bohlen, Johann; Salem, Clemens-Bässem; Tinnefeld, Philip; Ploetz, Evelyn & Lamb, Don C.
  
• Multicolor 3D Orbital Tracking. Small, 19(17).
  Mieskes, Frank; Ploetz, Evelyn; Wehnekamp, Fabian; Rat, Virgile & Lamb, Don C.
  
• Water Harvesting at the Single-Crystal Level. Journal of the American Chemical Society, 145(26), 14324-14334.
  Fuchs, Adrian; Knechtel, Fabian; Wang, Haoze; Ji, Zhe; Wuttke, Stefan; Yaghi, Omar M. & Ploetz, Evelyn
  
• Deep-LASI, single-molecule data analysis software. Biophysical Journal, 123(17), 2682-2695.
  Asadiatouei, Pooyeh; Salem, Clemens-Bässem; Wanninger, Simon; Ploetz, Evelyn & Lamb, Don C.