HAK-actin, a U-ExM-compatible probe to image the actin cytoskeleton

Area of Science:

• Cell Biology
• Microscopy
• Biochemistry

Background:

• Expansion microscopy (ExM) is a super-resolution technique that physically enlarges biological samples.
• While ExM excels at visualizing microtubules, effective probes for actin filaments are limited.
• Existing actin probes lack the robustness needed for diverse ExM applications.

Purpose of the Study:

• To develop and validate a novel probe for visualizing actin filaments using ultrastructure expansion microscopy (U-ExM).
• To provide a versatile and reliable tool for actin cytoskeleton imaging in various biological systems.

Main Methods:

• Engineered a new actin probe, termed HAK-actin.
• Tested HAK-actin compatibility with ultrastructure expansion microscopy (U-ExM).
• Evaluated HAK-actin performance in human cells, microbial eukaryotes, and mouse retinal tissue.

Main Results:

• HAK-actin demonstrated robust and uniform staining of actin filaments.
• The probe proved effective across diverse biological samples, including cellular and tissue contexts.
• HAK-actin facilitated high-resolution imaging of the actin cytoskeleton.

Conclusions:

• HAK-actin is a versatile and reproducible probe for actin visualization via U-ExM.
• This tool addresses a critical need for improved actin cytoskeleton imaging in cell biology.
• HAK-actin enables enhanced super-resolution microscopy of actin structures.