Multiplexed phase-space imaging for 3D fluorescence microscopy.

Area of Science:

• Optics and Photonics
• Microscopy
• Computational Imaging

Background:

• Optical phase-space functions capture spatial and angular information, crucial for applications like digital refocusing and 3D reconstruction.
• Existing methods for high-resolution 4D phase-space data acquisition are slow, light-inefficient, and lack diffraction-limited resolution.

Purpose of the Study:

• To introduce a novel multiplexed method for high-resolution 4D optical phase-space data capture.
• To overcome the limitations of speed, light efficiency, and resolution in previous scanning aperture techniques.

Main Methods:

• Utilizing a spatial light modulator (SLM) in the microscope's pupil plane to sequentially pattern multiplexed coded apertures.
• Capturing real-space images and reconstructing the 3D fluorescence distribution via regularized least squares with a proximal accelerated gradient descent solver.

Main Results:

• Experimental reconstruction of a 101 Megavoxel 3D volume (1010×510×500µm, NA 0.4).
• Demonstrated improvements in acquisition time, light throughput, and resolution compared to traditional scanning aperture methods.
• Showcased a flexible patterning scheme enabling exploitation of sample sparsity for reduced data capture.

Conclusions:

• The proposed multiplexed method offers a significant advancement for high-resolution 4D optical phase-space measurements.
• This technique provides a faster, more light-efficient, and higher-resolution alternative to existing scanning aperture approaches.
• The method's flexibility allows for optimized data acquisition based on sample characteristics.