In this paper, we present large-field, five-step lattice structured illumination microscopy (Lattice SIM). This method utilizes a 2D grating for lattice projection and a spatial light modulator (SLM) for phase shifting. Five phase-shifted intensity images are recorded to reconstruct a super-resolution image, enhancing the imaging speed and reducing the photo-bleaching both by 17%, compared to conventional two-direction and three-shift SIM. Furthermore, lattice SIM has a three-fold spatial bandwidth product (SBP) enhancement compared to SLM/DMD-based SIM, of which the fringe number is limited by the SLM/DMD pixel number. We believe that the proposed technique will be further developed and widely applied in many fields.

Structured illumination microscopy (SIM) has been widely used in biological research due to its merits of fast imaging speed, minimal invasiveness, super-resolution, and optical sectioning imaging capability. However, the conventional SIM that uses a spatial light modulator (SLM) for fringe projection often has a limited imaging field of view. Herein, we report a large-field SIM technique that combines a 2D grating for fringe pattern projection and an SLM for selecting fringe orientation and performing phase shifting digitally. The proposed SIM technique breaks the bottleneck of fringe number limited by the digital projection devices, while maintaining the advantage of high-speed (digital) phase shifting of conventional SIM. The method avoids the pixilation and dispersion effects of the SLMs. Finally, a 1.8-fold resolution enhancement in a large field of 690 × 517 µm² under a 20×/NA0.75 objective is experimentally demonstrated. The proposed technique can be widely applied to biology, chemistry, and industry.

Optical manipulation with nondiffracting beams has been attracting great interest and finding widespread applications in many fields such as chemistry, physics, and biomedicine. Generally, optical manipulation is conducted in an optical microscopy system, which, in general, only allows for imaging motions of particles in the transverse plane, rendering the observation of dynamics processes occurring in the axial plane impractical. We propose and demonstrate an optical manipulation system that incorporates an axial plane imaging module. With this system, the trapping behavior in the transverse plane and the transportation process in the axial plane of a particle immersed in a Bessel beam were acquired simultaneously in real time.

This study presents a dual-modality microscopic imaging approach that combines quantitative phase microscopy and fluorescence microscopy based on structured illumination (SI) to provide structural and functional information for the same sample. As the first imaging modality, structured illumination digital holographic microscopy (SI-DHM) is implemented along the transmission beam path. SI-DHM acts as a label-free, noninvasive approach and provides high-contrast and quantitative phase images utilizing the refractive index contrast of the inner structures of samples against the background. As the second imaging modality, structured illumination (fluorescence) microscopy (SIM) is constructed along the reflection beam path. SIM utilizes fluorescent labeling and provides super-resolution images for specific functional structures of samples. We first experimentally demonstrated phase imaging of SI-DHM on rice leaves and fluorescence (SIM) imaging on mouse kidney sections. Then, we demonstrated dual-modality imaging of biological samples, using DHM to acquire the overall cell morphology and SIM to obtain specific functional structures. These results prove that the proposed technique is of great importance in biomedical studies, such as providing insight into cell physiology by visualizing and quantifying subcellular structures.

Counting and analyzing of blood cells, as well as their subcellular structures, are indispensable for understanding biological processes, studying cell functions, and diagnosing diseases. In this paper, we combine digital holographic microscopy with cell segmentation guided by the Sobel operator using Dice coefficients for automatic threshold selection and aimed to automatic counting and analysis of blood cells in flow and different kinds of cells in the static state. We demonstrate the proposed method with automatic counting and analyzing rat red blood cells (RBCS) flowing in a microfluidic device, extracting quickly and accurately the size, concentration, and dry mass of the sample in a label-free manner. The proposed technique was also demonstrated for automatic segmentation of different cell types, such as COS7 and Siha. This method can help us in blood inspection, providing pathological information in disease diagnosis and treatment.