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Versatile Microfluidics Platform for Enhanced Multitarget Super-Resolution Microscopy
Samrat Basak1,2, Kim-Chi Vu3,4,5, Nikolaos Mougios6,7
1III. Institute of Physics - Biophysics, Georg August University, 37077 Göttingen, Germany.
ACS Nano
|January 1, 2026
Summary
We developed a microfluidics system for DNA-PAINT super-resolution microscopy, improving multiplexed imaging efficiency and reproducibility. This automated platform enables nanoscale imaging in complex biological samples like cells.
Area of Science:
- Super-resolution microscopy
- Nanoscale imaging
- Biophysics
Background:
- DNA-based Point Accumulation for Imaging in Nanoscale Topography (DNA-PAINT) is a single-molecule localization microscopy (SMLM) technique.
- Exchange-PAINT enables multiplexing by sequential target imaging using orthogonal DNA strands.
- Manual Exchange-PAINT workflows suffer from inefficiency, drift, variability, and poor reproducibility.
Purpose of the Study:
- To develop an automated, microfluidics-based system for enhanced multiplexed SMLM.
- To improve the efficiency, reproducibility, and material usage of Exchange-PAINT.
- To enable reliable nanoscale imaging in complex biological systems.
Main Methods:
- A custom compressed-air-driven microfluidics system with a stackable, modular design.
- Robust and material-efficient buffer exchange with minimal dead volume.
- Adaptable to various SMLM platforms (wide-field, confocal, STED, MINFLUX).
Main Results:
- Demonstrated robust 5-plex Exchange-PAINT imaging in U2OS cancer cells.
- Successfully performed multiplexed nanoscale imaging in fragile primary cardiomyocytes.
- The system ensures reproducible buffer exchange and minimal dead volume.
Conclusions:
- The microfluidics platform enables reliable super-resolution multiplexing in physiologically relevant systems.
- Facilitates detailed nanoscale analysis in complex primary cells.
- Offers a versatile solution for advanced SMLM applications.
