We investigate the diffraction effects of focused Gaussian beams yielding a double optical vortex by a nano-step structure fabricated in a transparent media. When approaching such a step the double vortex splits into single ones which move in a characteristic way. By observing this movement we can determine the position of the step with high resolution. Our theoretical predictions were verified experimentally.

Cancer treatment based on hyperthermia (HT) relies on exposing the malignant cells to elevated local temperature. Although the procedure has been successfully applied in clinics, the fundamental aspects of HT are not yet fully understood. In order to verify the susceptibility of single cells in vitro to raised temperature, we have developed novel nano- and microtools. In particular, an optical double-trap system utilizing combined galvano-mirror scanning and spatial light phase modulator was devised to manipulate several micron-sized objects simultaneously. The manipulation comprised both optical trapping and translocating, on demand photoactivated heating, and simultaneous remote temperature readout of living cells, infrared activated heaters and optical thermometers, respectively. Mesoporous silicon microparticles were used as an infrared absorber to generate an increased temperature of about 100 °C with 0.4 W laser power. The optical micron-sized thermometer was based on up-converting Yb–Er codoped nanocrystalline particles encapsulated in amorphous silica shells produced with yeast cells as the templates. These hybrid particles displayed a relative sensitivity of 0.28%/K, an accuracy of 0.1 °C (at 32 °C), spatial resolution of <10 μm, and a temporal response of 50 ms/acquisition to record the temperature changes in specified areas in real time. The system was utilized in monitoring the stepwise cell death of individual diffuse large B-cell lymphoma (DLBCL) cells due to locally induced excessive heating induced by the absorber localized in the vicinity of the cell.

Adhesion is critical for the maintenance of cellular structures as well as intercellular communication, and its dysfunction occurs prevalently during cancer progression. Recently, a growing number of studies indicated the ability of oxygen to regulate adhesion molecules expression, however, the influence of physiological hypoxia (physioxia) on cell adhesion remains elusive. Thus, here we aimed: (i) to develop an optical tweezers based assay to precisely evaluate single diffuse large B-cell lymphoma (DLBCL) cell adhesion to neighbor cells (mesenchymal stromal cells) and extracellular matrix (Matrigel) under normoxia and physioxia; and, (ii) to explore the role of integrins in adhesion of single lymphoma cell. We identified the pronouncedly reduced adhesive properties of lymphoma cell lines and primary lymphocytes B under physioxia to both stromal cells and Matrigel. Corresponding effects were shown in bulk adhesion assays. Then we emphasized that impaired β1, β2 integrins, and cadherin-2 expression, studied by confocal microscopy, account for reduction in lymphocyte adhesion in physioxia. Additionally, the blockade studies conducted with anti-integrin antibodies have revealed the critical role of integrins in lymphoma adhesion. To summarize, the presented approach allows for precise confirmation of the changes in single cell adhesion properties provoked by physiological hypoxia. Thus, our findings reveal an unprecedented role of using physiologically relevant oxygen conditioning and single cell adhesion approaches when investigating tumor adhesion in vitro.

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We have adapted a non-invasive method based on optical tweezers technology to differentiate between the normal B-cells and the B-cell non-Hodgkin lymphoma (B-NHL) cells derived from clinical samples. Our approach bases on the nascent adhesion between an individual B-cell and a mesenchymal stromal cell. In this study, a single B-cell was trapped and optically seeded on a mesenchymal stromal cell and kept in a direct contact with it until a stable connection between the cells was formed in time scale. This approach allowed us to avoid the introduction of any exogenous beads or chemicals into the experimental setup which would have affected the cell-to-cell adhesion. Here, we have provided new evidence that aberrant adhesive properties found in transformed B-cells are related to malignant neoplasia. We have demonstrated that the mean time required for establishing adhesive interactions between an individual normal B-cell and a mesenchymal stromal cell was 26.7 ± 16.6 s, while for lymphoma cell it was 208.8 ± 102.3 s, p < 0.001. The contact time for adhesion to occur ranged from 5 to 90 s and from 60 to 480 s for normal B-cells and lymphoma cells, respectively. This method for optically controlled cell-to-cell adhesion in time scale is beneficial to the successful differentiation of pathological cells from normal B-cells within the fine needle aspiration biopsy of a clinical sample. Additionally, variations in time-dependent adhesion among subtypes of B-NHL, established here by the optical trapping, confirm earlier results pertaining to cell heterogeneity.

We study the statistical properties of recordings that contain time-dependent positions of a bead trapped in optical tweezers. Analysis of such a time series indicates that the commonly accepted model, i.e., the autoregressive process of first-order, is not sufficient to fit the data. We show the presence of a first-order moving average part in the dynamical model of the system. We explain the origin of this part as an influence of the high-frequency CCD camera on the measurements. We show that this influence evidently depends on the applied exposure time. The proposed autoregressive moving average model appears to reflect perfectly all statistical features of the high-frequency recording data.

In the optical vortex microscopy the focused Gaussian beam with optical vortex scans a sample. An optical vortex can be introduced into a laser beam with the use of a special optical element--a vortex lens. When moving the vortex lens, the optical vortex changes its position inside the spot formed by a focused laser beam. This effect can be used as a new precise scanning technique. In this paper, we study the optical vortex behavior at the sample plane. We also estimate if the new scanning technique results in observable effects that could be used for a phase object detection.

Holographic optical tweezers were used to show the interaction between a strongly focused laser beam and magnetic nanoparticles in ferrofluid. When the light intensity was high enough, magnetic nanoparticles were removed from the beam center and formed a dark ring. The same behavior was observed when focusing vortex or Bessel beams. The interactions between two or more separated rings of magnetic nanoparticles created by independent optical traps were also observed.