Two-photon microscopy of acoustofluidic trapping for highly sensitive cell analysis

Area of Science:

• Biomedical Engineering
• Cell Biology
• Optical Microscopy

Background:

• Studying cells in vitro often lacks the physiological relevance of in vivo conditions.
• Existing methods may not provide high-resolution 3D analysis of cellular behavior in controlled environments.
• Investigating cell-cell interactions and functions requires techniques that minimize artificial constraints like wall contact.

Purpose of the Study:

• To develop and validate a novel 3D cell analysis system combining two-photon microscopy and acoustofluidic trapping.
• To enable in vitro studies of cells and cell clusters under conditions that closely mimic in vivo environments.
• To demonstrate the system's capability for real-time monitoring of cellular responses, such as osmosis.

Main Methods:

• Integration of two-photon-excited fluorescence microscopy with acoustofluidic trapping.
• Utilizing a spherical microchamber for three-dimensional sample manipulation and observation.
• High spatial precision and temporal resolution imaging for tracking dynamic cellular processes.

Main Results:

• Demonstrated in-depth 3D analysis of microchamber dimensions and precise positioning of trapped cells.
• Successfully monitored real-time osmosis in A549 lung cells and red blood cells.
• Observed significant reductions in cell membrane diameter during osmotic stress (approx. 4 μm for A549, 2 μm for RBCs).

Conclusions:

• The combined two-photon microscopy and acoustofluidic trapping system offers a powerful tool for 3D cell studies.
• This approach facilitates investigations of cellular functions and interactions in a near-physiological, wall-contact-free environment.
• The system is suitable for biomedical applications, including the study of osmotic responses in various cell types.