Correlative light- and electron microscopy with chemical tags

Area of Science:

• Cell Biology
• Microscopy Techniques
• Biochemistry

Background:

• Correlative microscopy combines light and electron microscopy for high-resolution imaging.
• Traditional methods often rely on green fluorescent protein (GFP) for light microscopy labeling.
• Limitations exist in achieving protein-specific, multicolor labeling with existing techniques.

Purpose of the Study:

• To develop a novel chemical tagging approach for correlative microscopy.
• To enable protein-specific and multicolor imaging using synthetic fluorophores.
• To improve the precision and applicability of correlative microscopy for complex biological samples.

Main Methods:

• Utilized chemical tagging with synthetic fluorophores for protein labeling.
• Performed multicolor imaging and super-resolution light microscopy.
• Applied identical sample preparation protocols for scanning electron microscopy (SEM) and transmission electron microscopy (TEM).

Main Results:

• Synthetic fluorophores maintained fluorescence after embedding and uranyl acetate treatment.
• Achieved precise superimposition of light and electron micrographs with up to 25 nm accuracy.
• Successfully visualized the molecular arrangement of cadherins in mouse epithelial cell adherens junctions.

Conclusions:

• Synthetic fluorophores offer a robust alternative to GFP for correlative microscopy.
• The new method provides high-quality, multicolor imaging with excellent preservation of fluorescence.
• This technique advances the capability to study intricate molecular arrangements in cellular structures.