3D super-resolution microscopy performance and quantitative analysis assessment using DNA-PAINT and DNA origami test...

Area of Science:

• Microscopy
• Biophysics
• Nanotechnology

Background:

• Accurate characterization of imaging setups is crucial for localization-based super-resolution techniques.
• Existing 2D imaging test samples are insufficient for specialized 3D and quantitative super-resolution methods.
• Robust and accessible test samples are needed for reliable performance feedback and method development.

Purpose of the Study:

• To introduce and analyze robust test samples for 3D and quantitative super-resolution imaging.
• To provide straightforward, cost-effective tools for assessing advanced microscopy performance.
• To facilitate the implementation of new super-resolution imaging methods.

Main Methods:

• Utilized microspheres functionalized for DNA-PAINT for 3D imaging quality assessment.
• Employed a commercial DNA origami sample for evaluating 3D imaging.
• Demonstrated a workflow for establishing and assessing quantitative PAINT (qPAINT) using a commercial DNA origami sample.

Main Results:

• Presented two viable options for assessing 3D super-resolution imaging quality.
• Showcased a method for establishing and evaluating qPAINT workflows for quantitative imaging.
• Confirmed the utility of the developed test samples for routine performance feedback.

Conclusions:

• Developed test samples enable robust assessment of 3D and quantitative super-resolution imaging.
• These samples are user-friendly, cost-effective, and require minimal specialized laboratory skills.
• The findings support broader adoption and reliable application of advanced super-resolution microscopy techniques.