A Non-Destructive, Tuneable Method to Isolate Live Cells for High-Speed AFM Analysis.

Area of Science:

• Biophysics
• Microscopy
• Cell Biology

Background:

• High-resolution imaging of single cells using atomic force microscopy (AFM) requires effective cell immobilization.
• Existing sample preparation methods may not withstand AFM forces or preserve sample integrity.

Purpose of the Study:

• To present a flexible methodology for rapid generation of bespoke arrays for cellular AFM analysis.
• To demonstrate the utility of multiphoton polymerization for cell immobilization.

Main Methods:

• Utilized high-resolution three-dimensional (3D) based printing via multiphoton polymerization.
• Generated custom arrays for immobilizing cells for AFM.
• Employed contact mode High-Speed Atomic Force Microscopy (HS-AFM) for imaging.

Main Results:

• Successfully generated bespoke arrays for cellular AFM analysis.
• Acquired high-resolution topographical data from live Emiliania huxleyi cells.
• Demonstrated continuous imaging of a single cell for over 90 minutes.

Conclusions:

• Multiphoton polymerization offers a rapid and flexible approach for cell immobilization in AFM studies.
• The developed methodology supports high-resolution imaging and mechanical property analysis of cells under physiological conditions.