Bruce Z. Gao

Generating radial or azimuthal polarization by axial sampling of circularly polarized vortex beams

A laser beam with circular polarization can be converted into either radial or azimuthal polarization by a microfabricated spiral phase plate and a radial (or azimuthal)-type linear analyzer. The resulting polarization is axially symmetric and is able to produce tightly focused light fields beyond the diffraction limit. We describe in detail the theory behind the technique and the experimental verification of the polarization both in the far field and at the focus of a high numerical aperture lens. Vector properties of the beam under strong focusing conditions were observed by comparing the fluorescence images corresponding to the focal intensity distribution for both radial and azimuthal polarizations. The technique discussed here may easily be implemented to a wide range of optical instruments and devices that require the use of tightly focused light beams.

Surface plasmon resonance imaging of cell-substrate contacts with radially polarized beams

We demonstrate the proof-of-concept for surface plasmon resonance sensing and imaging via a virtual probe at the cell-substrate interface of a biological cell in aqueous media. The technique is based on the optical excitation by focused radially polarized beams of localized surface plasmons, which forms a virtual probe on the metal substrate. The intensity distribution at the back focal plane of the objective lens enables quantitative measurements to be made of the cell-substrate contact. The acquired data is then visualized in the form of a local refractive index map.

Calibration of fringe projection profilometry with bundle adjustment strategy.

The measurement accuracy of fringe projection profilometry (FPP) largely depends on the calibration procedure. A more reliable calibration approach based on the stereo vision model of the FPP scheme in conjunction with the bundle adjustment strategy is presented. It can adjust the coordinates of benchmarks and thereby estimate the scheme parameters more accurately even with an imperfect target. The experiment results shows that the proposed approach can reach highly accurate calibration by solely using a printed target pattern, which verifies the proposed approach.

Universal and special keys based on phase-truncated Fourier transform

We propose a novel optical asymmetric cryptosystem based on a phase-truncated Fourier transform. Two decryption keys independent of each other are generated. They are referred to as universal key and special key, respectively. Each of them can be used for decryption independently in absence of the other. The universal key is applicable to decrypt any ciphertext encoded by the same encryption key, but with poor legibility. On the contrary, the special key is adequate for legible decryption, but only valid for one ciphertext corresponding to the specified plaintext. A set of simulation results show the interesting performance of two types of de cryption keys.

Laser-patterned stem-cell bridges in a cardiac muscle model for on-chip electrical conductivity analyses

Following myocardial infarction there is an irreversible loss of cardiomyocytes that results in the alteration of electrical propagation in the heart. Restoration of functional electrical properties of the damaged heart muscle is essential to recover from the infarction. While there are a few reports that demonstrate that fibroblasts can form junctions that transmit electrical signals, a potential alternative using the injection of stem cells has emerged as a promising cellular therapy; however, stem-cell electrical conductivity within the cardiac muscle fiber is unknown. In this study, an in vitro cardiac muscle model was established on an MEA-based biochip with multiple cardiomyocytes that mimic cardiac tissue structure. Using a laser beam, stem cells were inserted adjacent to each muscle fiber (cell bridge model) and allowed to form cell-cell contact as determined by the formation of gap junctions. The electrical conductivity of stem cells was assessed and compared with the electrical conductivities of cardiomyocytes and fibroblasts. Results showed that stem cell-myocyte contacts exhibited higher and more stable conduction velocities than myocyte-fibroblast contacts, which indicated that stem cells have higher electrical compatibility with native cardiac muscle fibers than cardiac fibroblasts.

Fringe projection 3D microscopy with the general imaging model.

Three-dimensional (3D) imaging and metrology of microstructures is a critical task for the design, fabrication, and inspection of microelements. Newly developed fringe projection 3D microscopy is presented in this paper. The system is configured according to camera-projector layout and long working distance lenses. The Scheimpflug principle is employed to make full use of the limited depth of field. For such a specific system, the general imaging model is introduced to reach a full 3D reconstruction. A dedicated calibration procedure is developed to realize quantitative 3D imaging. Experiments with a prototype demonstrate the accessibility of the proposed configuration, model, and calibration approach.

Mesenchymal Stem Cell-Cardiomyocyte Interactions under Defined Contact Modes on Laser-Patterned Biochips

Understanding how stem cells interact with cardiomyocytes is crucial for cell-based therapies to restore the cardiomyocyte loss that occurs during myocardial infarction and other cardiac diseases. It has been thought that functional myocardial repair and regeneration could be regulated by stem cell-cardiomyocyte contact. However, because various contact modes (junction formation, cell fusion, partial cell fusion, and tunneling nanotube formation) occur randomly in a conventional coculture system, the particular regulation corresponding to a specific contact mode could not be analyzed. In this study, we used laser-patterned biochips to define cell-cell contact modes for systematic study of contact-mediated cellular interactions at the single-cell level. The results showed that the biochip design allows defined stem cell-cardiomyocyte contact-mode formation, which can be used to determine specific cellular interactions, including electrical coupling, mechanical coupling, and mitochondria transfer. The biochips will help us gain knowledge of contact-mediated interactions between stem cells and cardiomyocytes, which are fundamental for formulating a strategy to achieve stem cell-based cardiac tissue regeneration.

Myosin filament assembly onto myofibrils in live neonatal cardiomyocytes observed by TPEF-SHG microscopy

AIMS Understanding myofibrillogenesis is essential for elucidating heart muscle formation, development, and remodelling in response to physiological stimulation. Here, we report the dynamic assembly process of contractile myosin filaments onto myofibrils in a live cardiomyocyte culture during myofibrillogenesis. METHODS AND RESULTS Utilizing a custom-built, two-photon excitation fluorescence and second harmonic generation imaging system equipped with an on-stage incubator, we observed new sarcomere additions in rat neonatal cardiomyocytes during 10 h of on-stage incubation. The new sarcomere additions occurred at the side of existing myofibrils, where we observed mature myofibrils acting as templates, or at the interstice of several separated myofibrils. CONCLUSIONS During sarcomeric addition, myosin filaments are assembled onto the premyofibril laterally. This lateral addition, which proceeds stepwise along the axial direction, plays an important role in the accumulation of Z-bodies to form mature Z-disks and in the regulation of sarcomeric alignment during maturation.

RECENT PROGRESS IN MULTIFOCAL MULTIPHOTON MICROSCOPY.

Multifocal multiphoton microscopy (MMM) has recently become an important tool in biomedicine for performing three-dimensional fast fluorescence imaging. Using various beamsplitting techniques, MMM splits the near-infrared laser beam into multiple beamlets and produces a multifocal array on the sample for parallel multiphoton excitation and then records fluorescence signal from all foci simultaneously with an area array detector, which significantly improves the imaging speed of multiphoton microscopy and allows for high efficiency in use of the excitation light. In this paper, we discuss the features of several MMM setups using different beamsplitting devices, including a Nipkow spinning disk, a microlens array, a set of beamsplitting mirrors, or a diffractive optical element (DOE). In particular, we present our recent work on the development of an MMM using a spatial light modulator (SLM).

Laser-guidance-based cell deposition microscope for heterotypic single-cell micropatterning

Cell patterning methods enable researchers to control specific homotypic and heterotypic contact-mediated cell-cell and cell-ECM interactions and to impose defined cell and tissue geometries. To micropattern individual cells to specific points on a substrate with high spatial resolution, we have developed a cell deposition microscope based on the laser-guidance technique. We discuss the theory of optical forces for generating laser guidance and the optimization of the optical configuration (NA ≈ 0.1) to manipulate cells with high speed in three dimensions. Our cell deposition microscope is capable of patterning different cell types onto and within standard cell research devices and providing on-stage incubation for long-term cell culturing. Using this cell deposition microscope, rat mesenchymal stem cells from bone marrow were micropatterned with cardiomyocytes into a substrate microfabricated with polydimethylsiloxane on a 22 mm × 22 mm coverglass to form a single-cell coculturing microenvironment, and their electrophysiological property changes were investigated during the coculturing days.