Structured illumination microscopy (SIM) achieves doubled spatial resolution through exciting the specimen with high-contrast, high-frequency sinusoidal patterns. Such an illumination pattern can be generated by laser interference or incoherent structured pattern. Opto-electronic devices, such as Spatial Light Modulator (SLM) or Digital Micro-mirror Device (DMD), can provide rapid switch of illumination patterns for SIM. Although DMD is much more cost-effective than SLM, it was previously restricted in association with incoherent light sources. To extend its application with coherent illumination, we model the DMD as a blazed grating, and simulate the effect with DMD pattern changes in SIM. Based on the simulation, we report a fast, high-resolution and cost-efficient SIM with DMD. Our home-built laser interference-based DMD-SIM (LiDMD-SIM) reveals the nuclear pore complex and microtubule in mammalian cells with doubled spatial resolution.
This work is a result of an exploration in response to the following observation: though reconstruction methodology of SIM image since its origin is carried out in frequency domain, recent works in this field, specially when it comes to SIM reconstruction using lesser than 9 raw images, have ventured into reconstruction methodologies that operate directly in the spatial domain. This work formulates and demonstrates a frequency domain reconstruction of SIM image using four raw images – one wide-field image and three SIM images. One of the chief feature of the presented reconstruction algorithm is that it employs the standard ‘tools and tricks’ used by the conventional 9-frame SIM reconstruction algorithm. Results indicate that the presented reconstruction algorithm provides high resolution reconstructions successfully as long as the noise level in the raw images is lower than 10%. For higher level of noise, the reconstruction result shows little resolution enhancement.
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