Combining atomic force-fluorescence microscopy with a stretching device for analyzing mechanotransduction processes...

Area of Science:

• Cell Biology
• Biophysics
• Biomaterials

Background:

• Mechanical forces significantly influence cellular behavior, impacting processes from normal function to disease.
• Understanding cellular responses to mechanical stress requires in vitro models with controlled stress application and immediate analytical feedback.

Purpose of the Study:

• To develop and validate a novel uniaxial motorized cell stretching device integrated with fluorescence microscopy (FM) and atomic force microscopy (AFM).
• To enable high-resolution live cell imaging and simultaneous mechanical analysis of cells under controlled mechanical stress.

Main Methods:

• Integration of a uniaxial motorized cell stretching device with a combined FM-AFM system.
• Application of reproducible uniaxial strain to cultured cells on polydimethylsiloxane (PDMS) membranes.
• Simultaneous fluorescence imaging and AFM measurements to analyze cytoskeletal networks and topographical changes.

Main Results:

• The system successfully applied physiologically relevant to hyperphysiological strain levels to cells.
• AFM measurements revealed topographical changes and alterations in mechanical characteristics, such as Young's modulus, in response to mechanical stress.
• A ~20% cell deformation induced observable changes in the cytoskeletal network's Young's modulus.

Conclusions:

• The integrated stretching device and FM-AFM system provide a powerful tool for dynamic analysis of cellular structural remodeling and mechanical properties under stress.
• This technology facilitates fundamental research into mechanotransduction pathways and cellular responses to mechanical stimuli.