Expansion microscopy with ninefold swelling (NIFS) hydrogel permits cellular ultrastructure imaging on conventional...

Area of Science:

• Cell Biology
• Microscopy
• Biotechnology

Background:

• Superresolution microscopy offers high-resolution cellular imaging but requires specialized equipment and complex processing.
• Expansion microscopy (ExM) enhances conventional microscopy resolution to ~70 nm by physically expanding specimens.
• Current ExM resolution (~70 nm) is still insufficient for resolving fine cellular ultrastructures compared to superresolution techniques (~30 nm).

Purpose of the Study:

• To develop a method that significantly improves the resolution of expansion microscopy using conventional microscopes.
• To achieve superresolution imaging capabilities accessible with standard laboratory equipment.
• To enable detailed visualization of cellular ultrastructures previously unresolvable by traditional microscopy.

Main Methods:

• Development of a novel ninefold swelling (NIFS) hydrogel for enhanced specimen expansion.
• Integration of experimental operations onto a detachable chip to ensure reproducibility.
• Application of the NIFS hydrogel and chip system for high-resolution imaging of cellular structures.

Main Results:

• Achieved a resolution of 31 nm using expansion microscopy with the NIFS hydrogel and conventional microscopy.
• Demonstrated the technique's efficacy on imaging challenging cellular ultrastructures like nuclear pore complexes and clathrin-coated pits.
• The integrated chip design facilitated maximal reproducibility of the high-resolution imaging process.

Conclusions:

• The NIFS hydrogel and integrated chip system provide a universal platform for superresolution imaging.
• This method significantly enhances the resolution of expansion microscopy, making superresolution accessible with standard microscopes.
• The technology enables detailed unveiling of cellular ultrastructural details, advancing biological research.