Thermal nanoimprint lithography for drift correction in super-resolution fluorescence microscopy

Area of Science:

• Biophysics
• Microscopy
• Nanotechnology

Background:

• Localization-based super-resolution microscopy achieves high accuracy but is limited by long acquisition times and stage drift.
• Previous UV-nanoimprint lithography (UV-NIL) fiducial markers had issues with autofluorescence, stability, and biocompatibility.

Purpose of the Study:

• To develop an improved fiducial micro-pattern for accurate 3D drift correction in super-resolution microscopy.
• To enhance biocompatibility, reduce autofluorescence, and improve structural stability compared to previous methods.

Main Methods:

• Fabrication of fiducial micro-patterns using thermal nanoimprint lithography (T-NIL).
• Utilizing a thermal plastic material with low autofluorescence and robust stability.
• Demonstrating drift correction capabilities with high-precision measurements.

Main Results:

• Achieved drift correction precision of 1.5 nm (lateral) and 6.1 nm (axial) every 0.2 seconds.
• The new T-NIL fiducial pattern exhibits low autofluorescence, especially in the blue region.
• Demonstrated high biocompatibility, ensuring cell viability and adhesion.

Conclusions:

• T-NIL offers a superior method for creating fiducial markers for super-resolution microscopy drift correction.
• The improved markers enable high-precision imaging of live and fixed biological samples.
• This technology advances the capability for detailed observation of cellular structures and dynamics.