Independent and simultaneous three-dimensional optical trapping and imaging.

Area of Science:

• Biophysics
• Optical Physics
• Materials Science

Background:

• Precise manipulation of micron-sized objects in 3D is crucial for fabricating advanced materials and studying cellular dynamics.
• Existing techniques often face limitations in simultaneous manipulation and high-resolution imaging.
• 3D holographic optical tweezers offer potential for multi-object manipulation, but integration with advanced imaging is key.

Purpose of the Study:

• To develop and demonstrate a combined 3D holographic optical tweezers and spinning-disk confocal microscopy system.
• To achieve high-accuracy manipulation and multi-channel fluorescence imaging of multiple micron-scaled objects.
• To explore applications in colloidal assembly and live cell dynamics studies.

Main Methods:

• Integration of dual-objective 3D holographic optical tweezers with a spinning-disk confocal microscope.
• Characterization of trapping accuracy (8 nm lateral, 20 nm axial) and precision (20 nm lateral, 200 nm axial).
• Simultaneous multi-channel fluorescence imaging during optical trapping.

Main Results:

• Demonstrated fabrication of ordered two-component and 3D colloidal arrays.
• Successfully trapped arrays of yeast cells for dynamic studies.
• Investigated yeast cell division kinetics within optical traps, revealing no alteration in division timescale.

Conclusions:

• The integrated system enables precise 3D manipulation and multi-channel imaging of micron-scale objects.
• The technology is suitable for fabricating complex colloidal structures and studying cellular processes.
• Optical trapping does not significantly impact the timescale of yeast cell division.