Aliaksandr V. Kachynski

Intense Visible and Near-Infrared Upconversion Photoluminescence in Colloidal LiYF4:Er3+ Nanocrystals under Excitation at 1490 nm

We report intense upconversion photoluminescence (PL) in colloidal LiYF(4):Er(3+) nanocrystals under excitation with telecom-wavelength at 1490 nm. The intensities of two- and three-photon anti-Stokes upconversion PL bands are higher than or comparable to that of the Stokes emission under excitation with low power density in the range 5-120 W/cm(2). The quantum yield of the upconversion PL was measured to be as high as ∼1.2 ± 0.1%, which is almost 4 times higher than the highest upconversion PL quantum yield reported to date for lanthanide-doped nanocrystals in 100 nm sized hexagonal NaYF(4):Yb(3+)20%, Er(3+)2% using excitation at ∼980 nm. A power dependence study revealed that the intensities of all PL bands have linear dependence on the excitation power density, which was explained by saturation effects in the intermediate energy states.

Elasticity-Mediated Self-Organization and Colloidal Interactions of Solid Spheres with Tangential Anchoring in a Nematic Liquid Crystal

Using laser tweezers, we study colloidal interactions of solid microspheres in the nematic bulk caused by elastic distortions around the particles with tangential surface anchoring. The interactions overcome the Brownian motion when the interparticle separation r-->p is less than 3 particle diameters. The particles attract when the angle theta between r-->p and the uniform far-field director n0 is between 0 degrees and approximately 70 degrees and repel when 75 degrees <or approximately theta<or=90 degrees. The particles aggregate in chains directed at approximately 30 degrees to n0 and, at higher concentrations, form complex kinetically trapped structures.

Nonlinear Optical Imaging and Raman Microspectrometry of the Cell Nucleus throughout the Cell Cycle

Fundamental understanding of cellular processes at molecular level is of considerable importance in cell biology as well as in biomedical disciplines for early diagnosis of infection and cancer diseases, and for developing new molecular medicine-based therapies. Modern biophotonics offers exclusive capabilities to obtain information on molecular composition, organization, and dynamics in a cell by utilizing a combination of optical spectroscopy and optical imaging. We introduce here a combination of Raman microspectrometry, together with coherent anti-Stokes Raman scattering (CARS) and two-photon excited fluorescence (TPEF) nonlinear optical microscopy, to study macromolecular organization of the nucleus throughout the cell cycle. Site-specific concentrations of proteins, DNA, RNA, and lipids were determined in nucleoli, nucleoplasmic transcription sites, nuclear speckles, constitutive heterochromatin domains, mitotic chromosomes, and extrachromosomal regions of mitotic cells by quantitative confocal Raman microspectrometry. A surprising finding, obtained in our study, is that the local concentration of proteins does not increase during DNA compaction. We also demonstrate that postmitotic DNA decondensation is a gradual process, continuing for several hours. The quantitative Raman spectroscopic analysis was corroborated with CARS/TPEF multimodal imaging to visualize the distribution of protein, DNA, RNA, and lipid macromolecules throughout the cell cycle.

Optical Trapping of Colloidal Particles and Measurement of the Defect Line Tension and Colloidal Forces in a Thermotropic Nematic Liquid Crystal

We demonstrate optical trapping and manipulation of transparent microparticles suspended in a thermotropic nematic liquid crystal with low birefringence. We employ the particle manipulation to measure line tension of a topologically stable disclination line and to determine colloidal interaction of particles with perpendicular surface anchoring of the director. The three-dimensional director fields and positions of the particles manipulated by laser tweezers are visualized by fluorescence confocal polarizing microscopy.

Laser trapping in anisotropic fluids and polarization-controlled particle dynamics

Anisotropic fluids are widespread, ranging from liquid crystals used in displays to ordered states of a biological cell interior. Optical trapping is potentially a powerful technique in the fundamental studies and applications of anisotropic fluids. We demonstrate that laser beams in these fluids can generate anisotropic optical trapping forces, even for particles larger than the trapping beam wavelength. Immersed colloidal particles modify the fluids ordered molecular structures and locally distort its optic axis. This distortion produces a refractive index “corona” around the particles that depends on their surface characteristics. The laser beam can trap such particles not only at their center but also at the high-index corona. Trapping forces in the beams lateral plane mimic the corona and are polarization-controlled. This control allows the optical forces to be reversed and cause the particle to follow a prescribed trajectory. Anisotropic particle dynamics in the trap varies with laser power because of the anisotropy of both viscous drag and trapping forces. Using thermotropic liquid crystals and biological materials, we show that these phenomena are quite general for all anisotropic fluids and impinge broadly on their quantitative studies using laser tweezers. Potential applications include modeling thermodynamic systems with anisotropic polarization-controlled potential wells, producing optically tunable photonic crystals, and fabricating light-controlled nano- and micropumps.

Coherent anti-Stokes Raman scattering polarized microscopy of three-dimensional director structures in liquid crystals

We demonstrate three-dimensional vibrational imaging of director structures in liquid crystals using coherent anti-Stokes Raman scattering (CARS) polarized microscopy. Spatial mapping of the structures is based on sensitivity of a polarized CARS signal to the orientation of anisotropic molecules in liquid crystals. As an example, we study structures in a smectic material and demonstrate that single-scan CARS and two-photon fluorescence images of molecular orientation patterns are consistent with each other and with the structure model.

Measurement of optical trapping forces by use of the two-photon-excited fluorescence of microspheres

A novel technique for the calibration of laser trapping systems that utilizes two-photon-excited fluorescence of commercial dye-stained microspheres has been demonstrated. The trapping forces as well as the trapping efficiency have been measured for various liquid environments and trapping depths. The trapping efficiency in water was found to decrease with an increase of trapping depths because of the enlargement of the trapping beam waist caused by aberrations of the optical system.

Biophotonic probing of macromolecular transformations during apoptosis

We introduce here multiplex nonlinear optical imaging as a powerful tool for studying the molecular organization and its transformation in cellular processes, with the specific example of apoptosis. Apoptosis is a process of self-initiated cell death, critically important for physiological regulation and elimination of genetic disorders. Nonlinear optical microscopy, combining the coherent anti-Stokes Raman scattering (CARS) microscopy and two-photon excited fluorescence (TPEF), has been used for analysis of spatial distribution of major types of biomolecules: proteins, lipids, and nucleic acids in the cells while monitoring their changes during apoptosis. CARS imaging revealed that in the nuclei of proliferating cells, the proteins are distributed nearly uniformly, with local accumulations in several nuclear structures. We have found that this distribution is abruptly disrupted at the onset of apoptosis and is transformed to a progressively irregular pattern. Fluorescence recovery after photobleaching (FRAP) studies indicate that pronounced aggregation of proteins in the nucleoplasm of apoptotic cells coincides with a gradual reduction in their mobility.

Realignment-enhanced coherent anti-Stokes Raman scattering and three-dimensional imaging in anisotropic fluids

We apply coherent anti-Stokes Raman Scattering (CARS) microscopy to characterize director structures in liquid crystals. We demonstrate that the polarized CARS signal in these anisotropic fluids strongly depends on alignment of chemical bonds/molecules with respect to the collinear polarizations of Stokes and pump/probe excitation beams. This dependence allows for the visualization of the bond/molecular orientations via polarized detection of the CARS signal and thus for CARS polarization microscopy of liquid crystal director fields, as we demonstrate using structures in nematic, cholesteric, and smectic liquid crystals. On the other hand, laser-induced director realignment at powers above a well-defined threshold provides the capability for all-optical CARS signal enhancement in liquid crystals. Moreover, since the liquid crystalline alignment can be controlled by electric and magnetic fields, this demonstrates the feasibility of CARS signal modulation by applying external fields to these materials.

Optical trapping of director structures and defects in liquid crystals using laser tweezers

We demonstrate optical manipulation of structures and defects in liquid crystals (LCs). The effective refractive index depends on the LC molecular orientations and the laser beams polarization. We use the orientation-mediated refractive index contrast for the laser trapping in LCs with a homogeneous composition, but with spatially-varying patterns of molecular orientations. Tightly-focused polarized beams allow for optical trapping of disclinations and their clusters, dislocations and oily streaks, cholesteric fingers and focal conic domains, etc. We calculate the optical gradient forces for typical structures and explain the trapping properties at low laser powers. We also show that when a high-power beam causes local molecular realignment, the laser trapping properties change for two reasons: (1) the refractive index pattern and optical gradient forces are modified; (2) additional elastic structural forces arise to minimize the elastic free energy.