An approach to achieve lateral superresolution for small probe confocal measurement system and its element

A shaped annular beam superresolution approach is proposed to improve a lateral resolution of a small probe laser confocal measurement system (LCMS). The approach proposed enables lateral superresolution measurement of LCMS to be achieved by using a binary optical diffractive element to shape a He–Ne Gaussian laser beam into an annular beam with an inner diameter of 0.87 mm and an outer diameter of 1.8 mm required for superresolution measurement, and shift the beam spatial frequency from low to high. And a binary optical element (BOE) with 16 phase levels is designed and fabricated to shape a Gaussian laser beam into an annular beam. Preliminary experimental results indicate that an intensity distribution of a shaped annular beam is in agreement with simulation results, the diffractive efficiency is 87.2%; LCMS lateral and axial resolutions of 0.2 μm and 3 nm are achieved, respectively, and its measurement range is expanded nearly to double, when BOE is used in LCMS and , NA=0.85.     

High-Precision Mapping of Membrane Proteins on Synaptic Vesicles using Spectrally Encoded Super-Resolution Imaging

The spatial resolution of single-molecule localization microscopy is limited by the photon number of a single switching event because of the difficulty of correlating switching events dispersed in time. Here we overcome this limitation by developing a new class of photoswitching semiconducting polymer dots (Pdots) with structured and highly dispersed single-particle spectra. We imaged the Pdots at the first and the second vibronic emission peaks and used the ratio of peak intensities as a spectral coding. By correlating switching events using the spectral coding and performing 4–9 frame binning, we achieved a 2–3 fold experimental resolution improvement versus conventional superresolution imaging. We applied this method to count and map SV2 and proton ATPase proteins on synaptic vesicles (SVs). The results reveal that these proteins are trafficked and organized with high precision, showing unprecedented level of detail about the composition and structure of SVs.     

用两区π-相位滤波器改善超半球形固体浸没透镜显微系统的成像特性

利用矢量衍射理论分析了π-相位滤波器在超半球形固体浸没透镜显微系统中的超分辨特性.结果显示,光斑的增益、Strehl率和边瓣强度随相位区域边界呈现出同步的、只一个周期的振荡性变化.从辐射场和速逝场的角度解释了这种变化的原因.对于近场显微,速逝场起决定性作用.π-相位滤波器的相位区域边界角大于临界角时,才能产生较大的超分辨作用.滤波器边界参量等于0.743（对应的角度大于临界角）的π-相位滤波器,可以使系统分辨率提高16%.     

Common-path phase-shifting digital holographic microscopy: A way to quantitative phase imaging and superresolution

We present an experimental setup useful for complex amplitude evaluation and phase image quantification of three-dimensional (3-D) samples in digital holographic microscopy (DHM). It is based on a common-path interferometric configuration performed by dividing the input plane in two contiguous regions and by placing a translation grating near to the Fourier plane. Then, complex amplitude distribution of the sample under test is recovered with phase-shifting standard method obtained by moving the grating using a linear motion stage. Some experimental results of an USAF resolution test are presented for different numerical aperture (NA) microscope lenses. In a second part, the proposed setup is tested under superresolution purposes. Based on the object’s spectrum shift produced by off-axis illumination, we use time multiplexing to generate a synthetic aperture enlargement that improves the final image resolution. Experimental results for the case of a biosample (human red blood cells) and a commercial low NA microscope lens validates the suggested superresolution approach.     

Designing superresolution optical pupil filter with constrained global optimization algorithm

In this paper, a new method for designing three-zone optical pupil filter is presented. The phase-only optical pupil filter and the amplitude-only optical pupil filters were designed. The first kind of pupil for optical data storage can increase the transverse resolution. The second kind of pupil filter can increase the axial and transverse resolution at the same time, which is applicable in three-dimension imaging in confocal microscopy.     

Photoactivatable Carbo- and Silicon-Rhodamines and Their Application in MINFLUX Nanoscopy

New photoactivatable fluorescent dyes (rhodamine, carbo- and silicon-rhodamines [SiR]) with emission ranging from green to far red have been prepared, and their photophysical properties studied. The photocleavable 2-nitrobenzyloxycarbonyl unit with an alpha-carboxyl group as a branching point and additional functionality was attached to a polycyclic and lipophilic fluorescent dye. The photoactivatable probes having the HaloTagTM amine (O2) ligand bound with a dye core were obtained and applied for live-cell staining in stable cell lines incorporating Vimentin (VIM) or Nuclear Pore Complex Protein NUP96 fused with the HaloTag. The probes were applied in 2D (VIM, NUP96) and 3D (VIM) MINFLUX nanoscopy, as well as in superresolution fluorescence microscopy with single fluorophore activation (VIM, live-cell labeling). Images of VIM and NUPs labeled with different dyes were acquired and their apparent dimensions and shapes assessed on a lower single-digit nanometer scale. Applicability and performance of the photoactivatable dye derivatives were evaluated in terms of photoactivation rate, labeling and detection efficiency, number of detected photons per molecule and other parameters related to MINFLUX nanoscopy.     

Theoretical investigation of optical patterning of monolayers with subwavelength resolution

We formulate a model of monolayer patterning via optically-controlled chemical reactions, with the goal of beating the diffraction limit in photolithography. We consider the use of the proven technique of STimulated Emission Depletion (STED) to selectively place a handful of molecules in a reactive excited state. We show that repeated optical excitation has a greater effect on pattern formation than increasing the reaction rate, auguring well for experimental work. We also consider optically-controlled deposition of a soluble species via STED, and show that even for very large concentrations and excited state lifetimes the full width at half maximum of the features formed is robust against the effects of diffusion and saturation.     

Transverse superresolution and focal shift with rotational tunable phase mask

This paper reports the imaging characteristics of an optical system can be modified by our designed polarization pupil mask. The novel rotational symmetric polarization pupil mask design based on combination of half-wave and quarter-wave plates is introduced for realizing the focal shift and extending focal depth of an optical system and the procedure for designing is presented. Numerical results show when an appropriate rotational symmetric polarization pupil mask is used as an apodizer in the optical imaging system, it not only can effectively achieve the continuously focal shift in a small range and extend focal depth of the optimized system, but also can evidently increase the transverse resolution of the optimized system at the genuine focal plane.     

Multichannel sampling expansion in the linear canonical transform domain and its application to superresolution

The linear canonical transform (LCT) describes the effect of first-order quadratic phase optical system on a wave field. In this paper, we address the problem of signal reconstruction from multichannel samples in the LCT domain based on a new convolution theorem. Firstly, a new convolution structure is proposed for the LCT, which states that a modified ordinary convolution in the time domain is equivalent to a simple multiplication operation for LCT and Fourier transform (FT). Moreover, it is expressible by a one dimensional integral and easy to implement in the designing of filters. The convolution theorem in FT domain is shown to be a special case of our achieved results. Then, a practical multichannel sampling expansion for band limited signal with the LCT is introduced. This sampling expansion which is constructed by the new convolution structure can reduce the effect of spectral leakage and is easy to implement. Last, the potential application of the multichannel sampling is presented to show the advantage of the theory. Especially, the application of multichannel sampling in the context of the image superresolution is also discussed. The simulation results of superresolution are also presented.