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New synthetic hydrogels for tumour spheroids and intestinal organoid cultures (publication)

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  We collaborated with groups of Prof. Richard Hoogenboom and Prof. Devriendt (Ghent University) and tested application of poly(2-alkyl-2-oxazoline) hydrogels (PAOx) for 3D cell cultures of cancer and stem cells. We found that encapsulation of small intestinal organoids into PAOx hydrogels results in polarity reversion, i.e. transition from basal to apical-out phenotype. This is useful for studying drug delivery, nutrient absorption and host-microbiota interactions within this model.  This figure shows dramatic differences of mitochondrial polarisation (TMRM) and appearance (transmission light) in small intestinal organoids, observed after apical-out transition (PAOx), in contrast to conventional Matrigel-grown culture: Since the PAOx hydrogels can show variable mechanical properties, we tested if encapsulation into stiff and soft hydrogels can affect behaviour of cancer cells in 3D: indeed, with HCT116 spheroids we found that stiffness of the hydrogels affect cell migration f...
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Happy New Year 2025! New publications.

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Wow, time really does fly! It's been a while since our previous update but by no means our team was idle.  End of 2024 was memorable with exciting conferences, including SPAOM 2024, Histochemical Society Annual online meeting 2024, FHERITALE consortium and others! We've received funding to study neurodevelopmental and neurodegenerative diseases (more news soon) so we can plan new & exciting research projects. Our team currently hosts 2 international researchers, Valerie Cote and Dr. Simone Perottoni, both learning and doing exciting science in FLIM, organoids and imaging of cancer cells / tissues. In addition, we are looking forward to host more of PhD students from FLIMagin3D consortium! Here are some recent publications, from our team and collaborative research projects: Sheehan A, Okkelman IA, Groslambert G, Bucher C, Dmitriev RI, Filatov MA: Optoelectronic Properties and Fluorescence Lifetime Imaging Application of Donor-Acceptor Dyads Derived From 2,6-DicarboxyBODIPY ...
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See the hidden: discover more with FLIM (online event)

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Prof. R. Dmitriev will participate in online event 'See the hidden: discover more with FLIM' on 10-Oct-2024! More info & registration can be found here: https://events.bitesizebio.com/see-the-hidden-discover-more-with/join
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Conference presentations: September 2024

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Our team is growing and we can present at the same time in two different places now! PhD candidate Nore Van Loon (joint project with FunGen lab, Prof. S. Vergult) gave a talk at the Methods and Applications of Fluorescence 2024 conference (8-11 September Valencia, Spain)! She presented her work in progress on using autofluorescence and fluorescent cholera toxin labelling to monitor differentiation of neural cells in human iPSC-derived 'brain' organoids. This work is a part of ongoing efforts of bringing together multi-omics approaches and advanced imaging of organoids to better understand aetiology of such neurodevelopmental disorders as FOXG1 syndrome, funded by the Special Research Fund at Ghent University (BOF) and Marguerite-Marie Delacroix foundation. At the same time, Prof. Ruslan Dmitriev travelled to Munster, Germany, to present an invited lecture on imaging oxygenation and cell metabolism in multicellular spheroids in 4th Inflammation & Imaging symposium! This l...
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Production and Multi-Parameter Live Cell Fluorescence Lifetime Imaging Microscopy (FLIM) of Multicellular Spheroids (JOVE protocol)

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ALERT: New publication from our group! Angela, Gabriele, Hang, Nore, Nina and Irina teamed up to produce a video protocol paper, focusing on comparing various multicellular spheroid formation methods, suitable for 'downstream' live multi-parameter fluorescence lifetime imaging microscopy (FLIM) and imaging cell oxygenation. This protocol is illustrated with examples of autofluorescence-based FLIM microscopy of NAD(P)H, oxygenation and cell death, as well as imaging of visible flavin-based cell autofluorescence, performed in tumour and stem cell-derived spheroids. The article is also a part of the Methods collection 'Quantitative Live Cell Imaging of 3D Models', which is going to expand in the future.    Link to the publication: https://www.jove.com/t/66845/production-multi-parameter-live-cell-fluorescence-lifetime-imaging The Methods Collection (Journal of Visualised Experiments, JOVE):  https://app.jove.com/methods-collections/902
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End of the Academic Year 2023-2024!

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It's been busy, it's been tough but it's been also fun! Our team has trained a number of MSc students, with excellent projects, spanning the scope from establishment of novel organoid models, to testing of mechanical properties of the hydrogel-producing biomaterials on the small intestinal epithelium organoids and FLIM analysis of neural ('brain') iPSC-derived organoids. Thanks to everyone involved! The end of year (June) was culminated in presenting of research at Ghent Gut Inflammation Group (Ruslan, Irina, Hang, Lenie) and participating (as instructors, Ruslan and Michele) in the FEBS course on Functional Imaging of cellular dynamics (Amsterdam, 9-15 June 2024)!         Stay tuned for more exciting news on our research!
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Brief updates for Academic Year of 2023-2024

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(video: time lapse FLIM of endocytosis in human colon cancer cells using LysoSensor Green, (C) Dmitriev Lab, 2023) The period of ~October 2023 until ~May 2024 (almost complete academic year!) was too busy for publishing any updates but thanks for staying tuned (if you did so). Some key updates for this period: Successful start of FLIMagin3D project, with the 3 new PhD students (Hang, Michele and Gabriele) + visiting (via secondments) PhD students, joining our team to address challenges in multi-parameter FLIM of organoids and multicellular spheroids, advancing imaging & probes for cell oxygenation, FLIM-FRET, tissue mechanics and biofabricated inter-species communities. More about the FLIMagin3D can be found here:  https://www.flimagin3d.com/   Some visiting scientists with exciting projects, including Dr. A. Russegger, R. Harkhoe, partners from OsteoNet Horizon MSCA Staff Exchange network Our group (including the microscopes!) has relocated to the new 'The Core' / MREB3 r...
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