High-accuracy differential autofocus system with an electrically tunable lens.

We propose a quasi-confocal microscopy autofocus system incorporating an electrically tunable lens (ETL) to achieve differential detection. The ETL changes its focal length to collect differential curves at speeds <300 Hz, allowing selective locking onto desired focal layers and high-speed differential operations close to the locked focal plane. By segmenting the system's pupil, the interference between the outgoing and incoming near-infrared beams is avoided, thereby greatly improving the signal-to-noise ratio. This ultra-sensitive system, with a focus drift accuracy better than 1/22 focal depth (∼20 nm @100× objective), provides a new, to the best of our knowledge, implementation pathway to meet the requirements of various microscopy techniques.

3D super-resolution microscopy based on nonlinear gradient descent structured illumination.

Three-dimensional structured illumination microscopy (3D-SIM) is an essential tool for volumetric fluorescence imaging, which improves both axial and lateral resolution by down-modulating high-frequency information of the sample into the passband of optical transfer function (OTF). And when combining with the 4Pi structure, the performance of 3D-SIM can be further improved. The reconstruction results of generally used linear 3D algorithm, however, are lack of high-fidelity and proneess to generate artifacts. In this paper, we proposed a novel iterative algorithm based on gradient descent combined with a nonlinear optimizer, which can be applied to all 3D-SIM setups (including I5S setup). We verified through simulation that the proposed solution, termed as nonlinear gradient descent structured illumination microscopy (NGD-SIM), achieves more fidelity results which can reach the limitation of theoretical resolution improvement of SIM. Moreover, it can be firmly validated on simulation that this algorithm can effectively reduce the amount of raw data in the case of sinusoidal-pattern illumination, i.e., the algorithm doesn't need five-step phase shifting; data with any number of phases can theoretically be reconstructed. Our method also provides the possibility to extend the application of sinusoidal-pattern illumination to any kind of interference fringe, which is generated by diversified types of illumination mode.

Fast reconstruction algorithm for structured illumination microscopy.

Structured illumination microscopy (SIM) is a powerful technique for providing super-resolution imaging, but its reconstruction algorithm, i.e., linear reconstruction structured illumination microscopy (LRSIM) algorithm in the Fourier domain, limits the imaging speed due to its computational effort. Here, we present a novel reconstruction algorithm that can directly process SIM data in the spatial domain. Compared to LRSIM, this approach uses the same number of frames to achieve a comparable resolution but with a much faster processing speed. Our algorithm was verified on both simulated and experimental data using sinusoidal pattern illumination. Moreover, this algorithm is also applicable for speckle pattern illumination.

Three-dimensional super-resolution imaging of live whole cells using galvanometer-based structured illumination microscopy.

Imaging and tracking three-dimensional (3D) nanoscale organizations and functions of live cells is essential for biological research but it remains challenging. Among different 3D super-resolution techniques, 3D structured illumination microscopy (SIM) has the intrinsic advantages for live-cell studies; it is based on wide-field imaging and does not require high light intensities or special fluorescent dyes to double 3D resolution. However, the 3D SIM system has developed relatively slowly, especially in live imaging. Here, we report a more flexible 3D SIM system based on two galvanometer sets conveniently controlling the structured illumination pattern's period and orientation, which is able to study dynamics of live whole cells with high speed. We demonstrate our microscope's capabilities with strong optical sectioning and lateral, axial, and volume temporal resolution of 104 nm, 320 nm and 4 s, respectively. We do this by imaging nanoparticle and microtubule organizations and mitochondria evolution. These characteristics enable our galvanometer-based 3D SIM system to broaden the accessible imaging content of SIM-family microscopes and further facilitate their applications in life sciences.

Surface wave illumination Fourier ptychographic microscopy.

We propose a novel microscopy method, combining surface wave illumination and the Fourier ptychographic microscopy (FPM) algorithm to achieve super-resolution (SR) imaging. In our system, an oil-immersion objective lens is used to excite both the total internal reflection (TIR) evanescent waves and the surface plasmon waves (SPWs), which illuminate the sample with large wave vectors. Through the FPM algorithm, a resolution approximately twice that of conventional wide-field microscopy is obtained. Meanwhile, we could retrieve the sample's quantitative phase map in order to obtain its surface profile. Importantly, the field enhancement from a SPW has improved the contrast of the reconstructed images, which is critical for revealing the finer structural details of the specimen. In our experiments, we have imaged metallic gratings with a 120 or 150 nm wide line and trench features. We accurately retrieved their axial dimensions with a lateral resolution better than 240 nm that is close to the theoretical resolution of 215 nm, thus demonstrating the quantitative phase imaging capability of our technique. As this approach provides a label-free solution for intensity and phase imaging of samples with lateral resolution exceeding the limit introduced by the optical system, it can be potentially used in a wide range of noninvasive biological imaging applications.

Light and matter co-confined multi-photon lithography.

Mask-free multi-photon lithography enables the fabrication of arbitrary nanostructures low cost and more accessible than conventional lithography. A major challenge for multi-photon lithography is to achieve ultra-high precision and desirable lateral resolution due to the inevitable optical diffraction barrier and proximity effect. Here, we show a strategy, light and matter co-confined multi-photon lithography, to overcome the issues via combining photo-inhibition and chemical quenchers. We deeply explore the quenching mechanism and photoinhibition mechanism for light and matter co-confined multiphoton lithography. Besides, mathematical modeling helps us better understand that the synergy of quencher and photo-inhibition can gain a narrowest distribution of free radicals. By using light and matter co-confined multiphoton lithography, we gain a 30 nm critical dimension and 100 nm lateral resolution, which further decrease the gap with conventional lithography.

Physicochemical and Functional Changes in Lotus Root Polysaccharide Associated with Noncovalent Binding of Polyphenols.

To promote the functional applications of lotus root polysaccharides (LRPs), the effects of noncovalent polyphenol binding on their physicochemical properties, as well as antioxidant and immunomodulatory activities, were investigated. Ferulic acid (FA) and chlorogenic acid (CHA) were spontaneously bound to the LRP to prepare the complexes LRP-FA1, LRP-FA2, LRP-FA3, LRP-CHA1, LRP-CHA2 and LRP-CHA3, and their mass ratios of polyphenol to LRP were, respectively, 121.57, 61.18, 34.79, 2359.58, 1276.71 and 545.08 mg/g. Using the physical mixture of the LRP and polyphenols as a control, the noncovalent interaction between them in the complexes was confirmed by ultraviolet and Fourier-transform infrared spectroscopy. The interaction increased their average molecular weights by 1.11~2.27 times compared to the LRP. The polyphenols enhanced the antioxidant capacity and macrophage-stimulating activity of the LRP depending on their binding amount. Particularly, the DPPH radical scavenging activity and FRAP antioxidant ability were positively related to the FA binding amount but negatively related to the CHA binding amount. The NO production of the macrophages stimulated by the LRP was inhibited by the co-incubation with free polyphenols; however, the inhibition was eliminated by the noncovalent binding. The complexes could stimulate the NO production and tumor necrosis factor-α secretion more effectively than the LRP. The noncovalent binding of polyphenols may be an innovative strategy for the structural and functional modification of natural polysaccharides.

Advances in the detection of ß-lactamase: A review.

ß-lactamase, an enzyme secreted by bacteria, is the main resistant mechanism of Gram-negative bacteria to ß-lactam antibiotics. The resistance of bacteria to ß-lactam antibiotics can be evaluated by testing the activity of ß-lactamase. Traditional phenotypic detection is a golden principle, but it is time-consuming. In recent years, many new methods have emerged, which improve the efficiency by virtue of their sensitivity, low cost, easy operation, and other advantages. In this paper, we systematically review these researches and emphasize their limits of detection, sample operation, and test duration. Noteworthily, some detection systems can identify the ß-lactamase subtype conveniently. We mainly divide these tests into three categories to elaborate their characteristics and application status. Both advantages and disadvantages of these methods are discussed. Additionally, we analyze the recent 5 years published researches to predict the trend of development in this field.

Twisted Optical Micro/Nanofibers Enabled Detection of Subtle Temperature Variation.

The detection of subtle temperature variation plays an important role in many applications, including proximity sensing in robotics, temperature measurements in microfluidics, and tumor monitoring in healthcare. Herein, a flexible miniaturized optical temperature sensor is fabricated by embedding twisted micro/nanofibers in a thin layer of polydimethylsiloxane. Enabled by the dramatic change of the coupling ratio under subtle temperature variation, the sensor exhibits an ultrahigh sensitivity (-30 nm/°C) and high resolution (0.0012 °C). As a proof-of-concept demonstration, a robotic arm equipped with our sensor can avoid undesired collisions by detecting the subtle temperature variation caused by the existence of a human. Moreover, benefiting from the miniaturized and engineerable sensing structure, real-time measurement of subtle temperature variation in microfluidic chips is realized. These initial results pave the way toward a category of optical sensing devices ranging from robotic skin to human-machine interfaces and implantable healthcare sensors.