Rifka Vlijm did her PhD in the group of Prof.dr. Cees Dekker at the Bionanoscience department of the technical university Delft. She here succeeded in assembling single nucleosomes onto DNA for single-molecule chromatin studies. In conjunction with dr. Yamini Dala at the National Cancer Institute (NIH, Bethesda, USA) she did an additional year as Postdoc in the same group to investigate the structural stability of centromere DNA. As these outcomes where contradicting existing models, she moved to the group of Nobel laureate Prof.dr. Stefan Hell, enabled by both an EMBL long-term fellowship (molecular biology) and a Rubicon grant (proposal ranked 1, physics). Here she built a Stimulated Emission Depletion (STED) microscopy, and developed the first protocols for live-cell STED microscopy on centromeres. In the year 2019 she joined the Zernike Institute for Advanced Materials as tenure-track assistant professor. Her group uses STED Microscopy to study cellular protein structures, especially those involved in cell division. An important part of the research focuses on microscopy developments enabling live-cell imaging: automation for high-throughput measurements and analysis, machine-learning enabled non-toxic imaging, and live-cell imaging protocols. In 2022 she received a personal NWO-M1 grant, a personal FSE research grant, and as consortium partner an NWO-XL grant. Furthermore, she was also nominated for the Aspasia price (outcome expected spring 2023). At the national level she is member of the advisory board of the NWO work council “Advanced Methods, Data, and Analyses to understand Living Systems” of the Life Science research community, and of the work council Physics for Life of the Physics community. Since 2021 she is member of the organizational committee of the NWO Dutch Biophysics, of which she is chair for the 2023 conference. She furthermore was involved in the initiation, and is currently board member of APNet, the national network for assistant professors.

Three top-publications:

- Hurtig et al. The patterned assembly and stepwise Vps4-mediated disassembly of composite ESCRT-III polymers drives archaeal cell division. Science Advances, 2023, accepted.

We used STED super-resolution microscopy to show how CdvB/CdvB1/CdvB2 proteins form a precisely patterned composite ESCRT-III division ring which undergoes stepwise Vps4-dependent disassembly and contracts to cut cells into two. These observations lead us to suggest sequential changes in a patterned composite polymer as a general mechanism of ESCRT-III-dependent membrane remodelling. Our novel high-throughput microscope for the first time allowed to apply STED to visualize cell division in wild-type archaea.

- de Lange, E.M.F. & Vlijm, R., Super-resolution imaging of peroxisomal proteins using STED nanoscopy. Methods in Molecular Biology, Springer Nature, 2023, accepted.

STED microscopy is a powerful technique enabling to reach resolutions of up to 27nm in cells using fluorescence microscopy. Although widely applied to e.g. mammalian cells, standard protocols for live cell yeast imaging were not yet developed. We here developed labelling and imaging methods to enable STED microscopy in living yeast cells, and provided guidelines for sample improvements, to make this technique accessible to a broader community.

- Vlijm, R., et al., STED nanoscopy of the centrosome linker reveals a CEP68-organized, periodic rootletin network anchored to a C-Nap1 ring at centrioles. Proc Natl Acad Sci U S A , 2018. 115(10): p. E2246 - E2253.

The centrosome linker plays important roles in cell organization, chromosome segregation, and cancer development. Too small for confocal light microscopy, and too sparse for labelling in electron microscopy, models on the centrosome linker were based on indirect measurements. We showed by STED (stimulated emission depletion microscopy) that the centrosome linker consists of a vast network of repeating rootletin units with a C-Nap1 ring at centrioles as organizer and CEP68 as filament modulator. We were even able to use optical microscopy to discriminate between the N- and C-terminus positions of the same protein, rootletin, enabling us to resolve the rootletin organization into filaments.