Abstract
.Significance: High-speed three-dimensional (3D) super-resolution microscopy is a unique tool to investigate various biological phenomena; yet the technology is not broadly adopted due to its high cost and complex system design.Aim: We present a compact, low-cost, and high-speed 3D structured illumination microscopy (SIM) based on a digital micromirror device and binary holography to visualize fast biological events with super-resolution.Approach: The 3D SIM uses a digital micromirror device to generate three laser foci with individually controllable positions, phases, and amplitudes via binary holography at the back aperture of objective lens to form optimal 3D structured patterns. Fifteen raw images are sequentially recorded and processed by the 3D SIM algorithm to reconstruct a super-resolved image.Results: Super-resolution 3D imaging at a speed of 26.7 frames per second is achieved with a lateral and axial resolution of 155 and 487 nm, which corresponds to a 1.65- and 1.63-times resolution enhancement, respectively, comparing with standard deconvolution microscopy.Conclusions: The 3D SIM realizes fast super-resolution imaging with optimal 3D structured illumination, which may find important applications in biophotonics.
Highlights
Structured illumination microscopy (SIM) is a fast and effective method to achieve superresolution imaging and optical cross-sectioning and has been widely used in biological studies
The 3D SIM realizes fast super-resolution imaging with optimal 3D structured illumination, which may find important applications in biophotonics
Fast 3D SIMs have been developed by replacing the grating with programmable spatial light modulators (SLM) or fast scanning devices;[3,4,5] for example, liquid crystal-based SLMs have been used in 3D SIM to generate high-resolution 3D structured patterns via phase modulations; notably, the temporal resolution is limited by the SLM pattern rate
Summary
Structured illumination microscopy (SIM) is a fast and effective method to achieve superresolution imaging and optical cross-sectioning and has been widely used in biological studies. The phase shifts and rotations of the illumination for frequency demodulations are achieved by mechanical translation and rotations of the grating, which requires careful tuning and has limited imaging speed. Fast 3D SIMs have been developed by replacing the grating with programmable spatial light modulators (SLM) or fast scanning devices;[3,4,5] for example, liquid crystal-based SLMs have been used in 3D SIM to generate high-resolution 3D structured patterns via phase modulations; notably, the temporal resolution is limited by the SLM pattern rate
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