Simple multi-color super-resolution by X10 microscopy.

Area of Science:

• Biophysics
• Microscopy
• Cell Biology

Background:

• Super-resolution microscopy techniques like STED, PALM, and STORM offer enhanced imaging but have technical limitations.
• Expansion microscopy (ExM) provides an alternative by physically expanding samples to overcome the diffraction limit.
• Previous ExM implementations achieved lower resolution (70-80nm) with a fourfold expansion factor.

Purpose of the Study:

• To introduce and evaluate the X10 expansion microscopy method for achieving higher resolution.
• To demonstrate the applicability of X10 ExM for multi-color imaging.
• To highlight X10 ExM as a promising tool for biological discovery.

Main Methods:

• Embedding biological samples within a swellable hydrogel.
• Isotropically expanding the gel-embedded sample to increase fluorophore separation.
• Utilizing standard fluorescence microscopes for imaging expanded samples.

Main Results:

• The X10 method achieves a tenfold expansion factor.
• This results in super-resolution imaging with a resolution of 25-30nm.
• X10 ExM is compatible with multi-color imaging and requires minimal technical setup.

Conclusions:

• X10 expansion microscopy breaks the diffraction barrier by physical sample expansion.
• It offers superior resolution compared to earlier ExM methods and is easier for multi-color imaging than other super-resolution techniques.
• X10 ExM is a powerful and accessible tool for advancing biological discoveries.