Imaging three-dimensional, subcellular structures with high axial resolution has always been the core purpose of fluorescence microscopy. However, trade-offs exist between axial resolution and other important technical indicators, such as temporal resolution, optical power density, and imaging process complexity. We report a new imaging modality, fluorescence interference structured illumination microscopy (FI-SIM), which is based on three-dimensional structured illumination microscopy for wide-field lateral imaging and fluorescence interference for axial reconstruction. FI-SIM can acquire images quickly within the order of hundreds of milliseconds and exhibit even 30 nm axial resolution in half the wavelength depth range without z-axis scanning. Moreover, the relatively low laser power density relaxes the requirements for dyes and enables a wide range of applications for observing fixed and live subcellular structures.
Direct laser writing (DLW) enables arbitrary three-dimensional nanofabrication. However, the diffraction limit poses a major obstacle for realizing nanometer-scale features. Furthermore, it is challenging to improve the fabrication efficiency using the currently prevalent single-focal-spot systems, which cannot perform high-throughput lithography. To overcome these challenges, a parallel peripheral-photoinhibition lithography system with a sub-40-nm two-dimensional feature size and a sub-20-nm suspended line width was developed in our study, based on two-photon polymerization DLW. The lithography efficiency of the developed system is twice that of conventional systems for both uniform and complex structures. The proposed system facilitates the realization of portable DLW with a higher resolution and throughput.
We present an alternative approach to realize structured illumination microscopy (SIM), which is capable for live cell imaging. The prototype utilizes two sets of scanning galvo mirrors, a polarization converter and a piezo-platform to generate a fast shifted, s-polarization interfered and periodic variable illumination patterns. By changing the angle of the scanning galvanometer, we can change the position of the spots at the pupil plane of the objective lens arbitrarily, making it easy to switch between widefield and total internal reflection fluorescent-SIM mode and adapting the penetration depth in the sample. Also, a twofold resolution improvement is achieved in our experiments. The prototype offers more flexibility of pattern period and illumination orientation changing than previous systems.
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