Jia Qian

Long-Distance Axial Trapping with Focused Annular Laser Beams

Focusing an annular laser beam can improve the axial trapping efficiency due to the reduction of the scattering force, which enables the use of a lower numerical aperture (NA) objective lens with a long working distance to trap particles in deeper aqueous medium. In this paper, we present an axicon-to-axicon scheme for producing parallel annular beams with the advantages of higher efficiency compared with the obstructed beam approach. The validity of the scheme is verified by the observation of a stable trapping of silica microspheres with relatively low NA microscope objective lenses (NA = 0.6 and 0.45), and the axial trapping depth of 5 mm is demonstrated in experiment.

Full-color structured illumination optical sectioning microscopy

In merits of super-resolved resolution and fast speed of three-dimensional (3D) optical sectioning capability, structured illumination microscopy (SIM) has found variety of applications in biomedical imaging. So far, most SIM systems use monochrome CCD or CMOS cameras to acquire images and discard the natural color information of the specimens. Although multicolor integration scheme are employed, multiple excitation sources and detectors are required and the spectral information is limited to a few of wavelengths. Here, we report a new method for full-color SIM with a color digital camera. A data processing algorithm based on HSV (Hue, Saturation, and Value) color space is proposed, in which the recorded color raw images are processed in the Hue, Saturation, Value color channels, and then reconstructed to a 3D image with full color. We demonstrated some 3D optical sectioning results on samples such as mixed pollen grains, insects, micro-chips and the surface of coins. The presented technique is applicable to some circumstance where color information plays crucial roles, such as in materials science and surface morphology.

Double-Exposure Optical Sectioning Structured Illumination Microscopy Based on Hilbert Transform Reconstruction

Structured illumination microscopy (SIM) with axially optical sectioning capability has found widespread applications in three-dimensional live cell imaging in recent years, since it combines high sensitivity, short image acquisition time, and high spatial resolution. To obtain one sectioned slice, three raw images with a fixed phase-shift, normally 2π/3, are generally required. In this paper, we report a data processing algorithm based on the one-dimensional Hilbert transform, which needs only two raw images with arbitrary phase-shift for each single slice. The proposed algorithm is different from the previous two-dimensional Hilbert spiral transform algorithm in theory. The presented algorithm has the advantages of simpler data processing procedure, faster computation speed and better reconstructed image quality. The validity of the scheme is verified by imaging biological samples in our developed DMD-based LED-illumination SIM system.

Image recombination transform algorithm for superresolution structured illumination microscopy

Abstract. Structured illumination microscopy (SIM) is an attractive choice for fast superresolution imaging. The generation of structured illumination patterns made by interference of laser beams is broadly employed to obtain high modulation depth of patterns, while the polarizations of the laser beams must be elaborately controlled to guarantee the high contrast of interference intensity, which brings a more complex configuration for the polarization control. The emerging pattern projection strategy is much more compact, but the modulation depth of patterns is deteriorated by the optical transfer function of the optical system, especially in high spatial frequency near the diffraction limit. Therefore, the traditional superresolution reconstruction algorithm for interference-based SIM will suffer from many artifacts in the case of projection-based SIM that possesses a low modulation depth. Here, we propose an alternative reconstruction algorithm based on image recombination transform, which provides an alternative solution to address this problem even in a weak modulation depth. We demonstrated the effectiveness of this algorithm in the multicolor superresolution imaging of bovine pulmonary arterial endothelial cells in our developed projection-based SIM system, which applies a computer controlled digital micromirror device for fast fringe generation and multicolor light-emitting diodes for illumination. The merit of the system incorporated with the proposed algorithm allows for a low excitation intensity fluorescence imaging even less than 1  W/cm2, which is beneficial for the long-term, in vivo superresolved imaging of live cells and tissues.

Accelerating beams with non-parabolic trajectories

We present a family of one-dimensional accelerating beams in the paraxial limit whose trajectory is described, in normalized coordinates (ξ, s), by a non-parabolic curve ξ = α(s) with α(s) denoting the transverse shift of beams. When taking different values of the parameters appearing in α(s), three types of accelerating beams are observed. The first type accelerates initially along X direction and almost stops accelerating after traveling a large distance. The second type is seen to travel along a straight line with an angle with respect to the Z-axis at large distance. The beam of the last type is seen to leave initially upward and cross the Z-axis downward after traveling some distance.

Three-dimensional characterization of tightly focused fields for various polarization incident beams

Tightly focused vectorial optical beams have found extensive applications in variety of technical fields like single-molecule detection, optical tweezers, and super-resolution optical microscopy. Such applications require an accurate measurement and manipulation of focal optical fields. We have developed a compact instrument (with dimensions of 35 × 35 × 30 cm3) to rapidly measure the intensity distribution in three dimensions of the focused fields of vectorial beams and any other incident beams. This instrument employs a fluorescent nanoparticle as a probe to scan the focal region to obtain a high spatial resolution of intensity distribution. It integrates a liquid-crystal spatial light modulator to allow for tailoring the point spread function of the optical system, making it a useful tool for multi-purpose and flexible research. The robust applicability of the instrument is verified by measuring the 3D intensity distributions of focal fields of various polarization and wavefront modulated incident beams focused by a high NA (=1.25) objective lens. The minimal data acquisition time achievable in the experiment is about 8 s for a scanning region of 3.2 × 3.2 μm2 (512 × 512 pixels). The measured results are in good agreement with those predicted by the vectorial diffraction theory.

Fast DMD based super-resolution structured illumination microscopy

Structured illumination microscopy (SIM) is an attractive choice for fast super-resolution imaging. The generation of structured illumination patterns made by interference of laser beams is broadly employed to obtain high modulation depth of patterns, while the polarizations of the laser beams must be elaborately controlled to guarantee the high contrast of interference intensity, which brings a more complex configuration for the polarization control. The emerging pattern projection strategy is much more compact, but the modulation depth of patterns is deteriorated by the optical transfer function (OTF) of the optical system, especially in high spatial frequency near diffraction limit. Therefore, the traditional super-resolution reconstruction algorithm for interference-based SIM will suffer from much artifact in the case of projection-based SIM that possesses a low modulation depth. Here, we propose an alternative reconstruction algorithm based on image recombination transform (IRT), which provides an alternative solution to address this problem even in a weak modulation depth. We demonstrated the effectiveness of this algorithm in the multicolor super-resolution imaging of BPAE cells in our developed projection-based SIM system, which applies a computer controlled digital micro-mirror device (DMD) for fast fringe generation and multicolor LEDs for illumination. The merit of the system incorporated with the proposed algorithm allows for a low excitation intensity fluorescence imaging even less than 1W/cm2, which is beneficial for the long-term, in vivo super-resolved imaging of live cells and tissues.