High-resolution imaging of cellular processes across textured surfaces using an indexed-matched elastomer

Area of Science:

• Bioengineering
• Cell Biology
• Biophysics

Background:

• Cell-environment interactions are crucial in bioengineering and stem cell research.
• Surface topography and rheology influence cell behavior but hinder high-resolution imaging.
• Existing methods struggle to provide detailed molecular insights into cellular environmental sensing.

Purpose of the Study:

• To develop a method for high-resolution imaging of cells on nano/microstructured surfaces.
• To enable simultaneous measurement of cellular forces and imaging of subcellular structures.
• To advance the study of cell-environment interactions and sensing mechanisms.

Main Methods:

• Molding nano/microstructured surfaces from an elastomeric material with a refractive index matched to cell culture medium.
• Rendering surfaces biocompatible for cell culture.
• Utilizing inverted microscopy for contrast (DIC, phase contrast) and fluorescence imaging through textured surfaces.
• Performing simultaneous traction force microscopy via micropost deflection.

Main Results:

• Successfully created biocompatible nano/microstructured surfaces compatible with high-resolution imaging.
• Enabled precise imaging of subcellular structures through textured substrates.
• Demonstrated simultaneous traction force measurements, correlating cellular forces with environmental interactions.
• Overcame imaging limitations previously associated with textured surfaces.

Conclusions:

• The developed technique allows for unprecedented resolution in studying cell-environment interactions.
• This approach facilitates a deeper understanding of cellular sensing processes and force generation mechanisms.
• It holds significant potential for next-generation in vitro assays and organs-on-chips.