Multiplexing Focused Ultrasound Stimulation with Fluorescence Microscopy.

Area of Science:

• Biomedical Engineering
• Cell Biology
• Acoustic Physics

Background:

• Low-intensity pulsed ultrasound (LIPUS) shows potential for non-invasive cellular manipulation.
• Understanding the biophysical mechanisms of LIPUS interaction with tissues is crucial for clinical translation.
• Current methods for studying LIPUS effects are limited by potential artifacts.

Purpose of the Study:

• To develop a method for integrating LIPUS with epi-fluorescence microscopy.
• To minimize acoustic artifacts introduced by the microscope setup.
• To investigate LIPUS-induced cellular responses, such as calcium transients.

Main Methods:

• A step-by-step procedure for incorporating a single-element ultrasound transducer into an upright epi-fluorescence microscope.
• Alignment of the ultrasound focal zone with the microscope objective focal point.
• Calcium imaging of cultured human glioblastoma cells exposed to LIPUS.

Main Results:

• Successful integration of LIPUS delivery into a standard fluorescence microscope setup.
• Minimization of acoustic wave reflections and refractions at interfaces.
• Observation of LIPUS-induced calcium transients in glioblastoma cells.

Conclusions:

• The described method allows for the study of LIPUS effects on cells using fluorescence microscopy.
• This technique facilitates the investigation of biophysical mechanisms underlying LIPUS cellular responses.
• Enables further research into LIPUS applications in neuroscience and regenerative medicine.