A versatile microfluidics platform for enhanced multi-target super-resolution microscopy Article Swipe
Roman Tsukanov
,
S. Basak
,
Kim Chi-Vu
,
Nikolaos Mougios
,
Nazar Oleksiievets
,
Yoav G. Pollack
,
Sören Brandenburg
,
Felipe Opazo
,
Stephan E. Lehnart
,
Jörg Enderlein
·
YOU?
·
· 2025
· Open Access
·
· DOI: https://doi.org/10.1101/2025.10.07.680900
YOU?
·
· 2025
· Open Access
·
· DOI: https://doi.org/10.1101/2025.10.07.680900
DNA-based Point Accumulation for Imaging in Nanoscale Topography (DNA-PAINT) is a powerful variant of single-molecule localization microscopy (SMLM) that overcomes the limitations of photobleaching, offers flexible fluorophore selection, and enables fine control of imaging parameters through tunable on- and off-binding kinetics. Its most distinctive feature is the capacity for multiplexing, which is achieved through a process known as Exchange-PAINT. This technique involves assigning orthogonal DNA strands to different targets within a sample and then sequentially adding and removing complementary imager strands that are specific to only one target at a time. However, manual Exchange-PAINT workflows are often inefficient, prone to drift and variability, and lack reproducibility. Here, we introduce a custom compressed-air-driven microfluidics system specifically designed for multiplexed SMLM. Featuring a stackable and modular design that is, in principle, not limited by the number of channels, the system ensures robust, reproducible, and material-efficient buffer exchange with minimal dead volume. It operates in both manual and automated modes and can be readily adapted to a wide range of commercial and custom microscopes, including wide-field, confocal, STED, and MINFLUX platforms. We demonstrate robust 5-plex Exchange-PAINT imaging in cancerous U2OS cells and, importantly, we establish multiplexed nanoscale imaging in fragile primary cardiomyocytes. These applications highlight the unique power of our platform to extend super-resolution multiplexing into physiologically relevant systems, thereby opening new avenues for biomedical research.
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Metadata
- Type
- preprint
- Language
- en
- Landing Page
- https://doi.org/10.1101/2025.10.07.680900
- https://www.biorxiv.org/content/biorxiv/early/2025/10/07/2025.10.07.680900.full.pdf
- OA Status
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- References
- 34
- OpenAlex ID
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All OpenAlex metadata
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https://openalex.org/W4414920769Canonical identifier for this work in OpenAlex
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https://doi.org/10.1101/2025.10.07.680900Digital Object Identifier
- Title
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A versatile microfluidics platform for enhanced multi-target super-resolution microscopyWork title
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preprintOpenAlex work type
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enPrimary language
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2025Year of publication
- Publication date
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2025-10-07Full publication date if available
- Authors
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Roman Tsukanov, S. Basak, Kim Chi-Vu, Nikolaos Mougios, Nazar Oleksiievets, Yoav G. Pollack, Sören Brandenburg, Felipe Opazo, Stephan E. Lehnart, Jörg EnderleinList of authors in order
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https://doi.org/10.1101/2025.10.07.680900Publisher landing page
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https://www.biorxiv.org/content/biorxiv/early/2025/10/07/2025.10.07.680900.full.pdfDirect link to full text PDF
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YesWhether a free full text is available
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greenOpen access status per OpenAlex
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https://www.biorxiv.org/content/biorxiv/early/2025/10/07/2025.10.07.680900.full.pdfDirect OA link when available
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0Total citation count in OpenAlex
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34Number of works referenced by this work
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