Robust and artifact-free mounting of tissue samples for atomic force microscopy

Area of Science:

• Biotechnology
• Materials Science
• Microscopy Techniques

Background:

• Traditional atomic force microscopy (AFM) sample preparation methods, such as semi-drying or glue-based immobilization, can introduce artifacts.
• These artifacts compromise the integrity and accuracy of AFM analysis, particularly for delicate biological tissues.

Purpose of the Study:

• To introduce and evaluate a novel Soft- Clamping Immobilizing Retainer of Tissue (SCIRT) system.
• To demonstrate the SCIRT method's effectiveness for nondestructive immobilization of soft, hydrated tissues for AFM analysis.

Main Methods:

• Design and implementation of the Soft- Clamping Immobilizing Retainer of Tissue (SCIRT).
• Comparative analysis of SCIRT versus traditional glue-immobilization techniques.
• Testing on challenging tissue samples: human trabecular meshwork (HTM) and rabbit cornea (RC).

Main Results:

• The SCIRT method enables the use of small sample sizes.
• It facilitates continuous sample hydration, crucial for biological integrity.
• Eliminates artifacts associated with glue and solvents, allowing for sample recovery post-measurement.

Conclusions:

• The SCIRT method offers a simple, effective, and artifact-free solution for immobilizing soft, hydrated tissues for AFM.
• It overcomes limitations of conventional methods, enhancing the reliability of AFM data for complex biological samples.