Baoli Yao

DMD-based LED-illumination Super-resolution and optical sectioning microscopy

Super-resolution three-dimensional (3D) optical microscopy has incomparable advantages over other high-resolution microscopic technologies, such as electron microscopy and atomic force microscopy, in the study of biological molecules, pathways and events in live cells and tissues. We present a novel approach of structured illumination microscopy (SIM) by using a digital micromirror device (DMD) for fringe projection and a low-coherence LED light for illumination. The lateral resolution of 90 nm and the optical sectioning depth of 120 μm were achieved. The maximum acquisition speed for 3D imaging in the optical sectioning mode was 1.6×107 pixels/second, which was mainly limited by the sensitivity and speed of the CCD camera. In contrast to other SIM techniques, the DMD-based LED-illumination SIM is cost-effective, ease of multi-wavelength switchable and speckle-noise-free. The 2D super-resolution and 3D optical sectioning modalities can be easily switched and applied to either fluorescent or non-fluorescent specimens.

Phase-shift extraction for generalized phase-shifting interferometry

A simple algorithm for blind extraction of phase shifts is proposed for generalized phase-shifting interferometry from only three interferograms. Based on the statistical property of the object wave, the algorithm calculates approximately the involved phase shifts as initial values. The extraction is further improved by an iterative method, considering the fact that the closer the phase shifts approach their real values, the more uniform the reconstructed reference wave will become. The feasibility of this algorithm is demonstrated by both simulation and experiment.

Polarization patterns hide and display using photoinduced anisotropy of photochromic fulgide

Photoinduced anisotropy of a photochromic pyrrylfulgide/PMMA film was investigated by using two linearly polarized beams. Excitation by linearly polarized light induces into the film an optical axis that has the same polarization as the excitation beam. This causes a change of the transmittance and of the polarization state of the detection beam. With a microscope a matrix of 4x4 light spots with different polarizations were recorded in the pyrrylfulgide/PMMA film. If readout with non-polarized light, the matrix of light spots show no information pattern. However, when readout with differently polarized lights, different patterns can be displayed. The experiment demonstrates that pyrrylfulgide/PMMA films can be used to hide two differently polarized patterns, which may be applied in camouflage technology.

Trapping of low-refractive-index particles with azimuthally polarized beam

Azimuthally polarized beams, focused by a high-numerical-aperture (NA) objective lens, form a hollow intensity distribution near the focus, which is appropriate for trapping low-refractive-index particles, in contrast to common linearly polarized or radially polarized beams. In this paper, the field distribution of the azimuthally polarized beam focused by a high-NA objective is described by the vectorial diffraction integral, and then the radiation forces on spherical particles with different parameters such as radius and refractive index are calculated by the T-matrix method. Numerical results show that the azimuthally polarized beam not only can steadily trap low-refractive-index particles at the focus center but also can trap multiple high-refractive-index particles around the focus center by virtue of the hollow-ring configuration. The range of the sizes of low-refractive-index particles that can be trapped steadily are presented, corresponding to different parameters such as the NA of the objective and the relative refractive index, based on which the NA of the objective can be selected to trap the appropriate size of particles

Parallel two-step phase-shifting point-diffraction interferometry for microscopy based on a pair of cube beamsplitters

Parallel two-step phase-shifting point-diffraction interferometry for microscopy based on a pair of cube beamsplitters is proposed. The first 45°-tilted cube beamsplitter splits object wave into two parallel copies: one copy is filtered by a pinhole in its Fourier plane to behave as reference wave, while the other one remains unchanged as object wave. The second cube beamsplitter combines the object and reference waves, and then split them together into two beams. Along with the two beams, two parallel phase-shifting interferograms are obtained in aid of polarization elements. Based on the proposed configuration, slightly-off-axis interferometry for microscopy is performed, which suppresses dc term by subtracting the two phase-shifting holograms from each other. The setup is highly stable due to its common-path configuration, and has been demonstrated to be suitable for measuring moving objects or dynamic processes.

Polarization holographic high-density optical data storage in bacteriorhodopsin film

Optical films containing the genetic variant bacteriorhodopsin BR-D96N were experimentally studied in view of their properties as media for holographic storage. Different polarization recording schemes were tested and compared. The influence of the polarization states of the recording and readout waves on the retrieved diffractive images intensity and its signal-to-noise ratio were analyzed. The experimental results showed that, compared with the other tested polarization relations during holographic recording, the discrimination between the polarization states of diffracted and scattered light is optimized with orthogonal circular polarization of the recording beams, and thus a high signal-to-noise ratio and a high diffraction efficiency are obtained. Using a He-Ne laser (633 nm, 3 mW) for recording and readout, a spatial light modulator as a data input element, and a 2D-CCD sensor for data capture in a Fourier transform holographic setup, a storage density of 2 x 10(8) bits/cm2 was obtained on a 60 x 42 microm2 area in the BR-D96N film. The readout of encoded binary data was possible with a zero-error rate at the tested storage density.

Autofocusing of digital holographic microscopy based on off-axis illuminations

An auto-focusing method for digital holographic microscopy has been proposed by employing two off-axis illumination beams. When specimens are illuminated by two plane waves in different directions, it is found that the farther the reconstruction plane is from the image plane, the wider the two reconstructed images are separated from each other. Thus, the image plane can be determinated by finding the minimum of the variation between the two reconstructed object waves on both the amplitude and phase distributions. The feasibility of the proposed method is demonstrated by the corresponding simulation and experiment.

Phase-shifting point-diffraction interferometry with common-path and in-line configuration for microscopy.

A new common-path and in-line point-diffraction interferometer for quantitative phase microscopy is proposed. The interferometer is constructed by introducing a grating pair into the point-diffraction interferometer, thus forming a common-path and in-line configuration for object and reference waves. Achromatic phase shifting is implemented by linearly moving one of the two gratings in its grating vector direction. The feasibility of the proposed configuration is demonstrated by theoretical analysis and experiments.

Autofocusing based on wavelength dependence of diffraction in two-wavelength digital holographic microscopy.

An autofocusing method for two-wavelength digital holographic microscopy (TWDHM) based on the wavelength dependence of the diffraction process is proposed. Red and green lights are employed for the illumination of the TWDHM, and the generated holograms are recorded simultaneously by a color CCD camera. Due to the wavelength dependency of the diffraction process, the farther the reconstruction plane is from the image plane, the larger the difference is between the red and green light distributions. Thus, the image plane can be determined by finding the minimum of the variation between the red and green lights on their amplitude distributions. The feasibility of the proposed method is demonstrated by simulation and experiment.

Dual-wavelength slightly off-axis digital holographic microscopy

We propose dual-wavelength digital holographic microscopy with a slightly off-axis configuration. The axial measurement range without phase ambiguity is extended to the micrometer range by synthesizing a beat wavelength between the two wavelengths with separation of 157 nm. Real-time measurement of the specimen is made possible by virtue of the high wavelength selectivity of the Bayer mosaic filtered color CCD camera. The principle of the method is exposed, and the practicability of the proposed configuration is demonstrated by the experimental results on a vortex phase plate and a rectangular phase step.