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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.

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|January 1, 2026
PubMed
Summary
This summary is machine-generated.

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.

Keywords:
DNA-PAINTSMLM automationcardiomyocytesmicrofluidicsmultiplexed imagingsingle-molecule localization microscopysuper-resolution microscopy

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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.