Quantitative imaging of cellular adhesion by total internal reflection holographic microscopy

Area of Science:

• Biophysics
• Cell Biology
• Microscopy

Background:

• Quantitative phase microscopy offers label-free imaging of biological samples.
• Total internal reflection (TIR) is a phenomenon used in various optical techniques.
• Cell-substrate interfaces are critical for understanding cell behavior and interactions.

Purpose of the Study:

• To present total internal reflection (TIR) holographic microscopy as a novel method for imaging cell-substrate interfaces.
• To demonstrate the capability of TIR holographic microscopy for quantitative phase imaging.
• To visualize microscopic structures at the cell-substrate interface.

Main Methods:

• Utilizing a prism in total internal reflection (TIR) as a near-field imager.
• Employing evanescent waves to probe phase shifts caused by refractive index variations.
• Performing quantitative phase microscopy on biological specimens.

Main Results:

• Generated quantitative phase images of test specimens including Amoeba proteus.
• Obtained quantitative phase images of SKOV-3 and 3T3 fibroblast cells.
• Demonstrated modulation of the evanescent wave phase front by cellular structures and adhesions.

Conclusions:

• Total internal reflection (TIR) holographic microscopy is effective for quantitative phase imaging of cell-substrate interfaces.
• The technique can visualize microscopic organisms, adhesions, and tissue structures.
• This method provides a new tool for studying cell-substrate interactions in biophysics and cell biology.