Kyoung Jin Lee

Competing patterns of signaling activity in dictyostelium discoideum.

Quantitative experiments are described on spatio-temporal patterns of coherent chemical signaling activity in populations of {it Dictyostelium discoideum} amoebae. We observe competition between spontaneously firing centers and rotating spiral waves that depends strongly on the overall cell density. At low densities, no complete spirals appear and chemotactic aggregation is driven by periodic concentric waves, whereas at high densities the firing centers seen at early times nucleate and are apparently entrained by spiral waves whose cores ultimately serve as aggregation centers. Possible mechanisms for these observations are discussed.

Full-field and single-shot quantitative phase microscopy using dynamic speckle illumination

We developed an off-axis quantitative phase microscopy that works for a light source with an extremely short spatial coherence length in order to reduce the diffraction noise and enhance the spatial resolution. A dynamic speckle wave whose coherence length is 440 nm was used as an illumination source. To implement an off-axis interferometry for a source of low spatial coherence, a diffraction grating was inserted in the reference beam path. In doing so, an oblique illumination was generated without rotation of the wavefront, which leads to a full-field and single-shot phase recording with improved phase sensitivity of more than a factor of 10 in comparison with coherent illumination. The spatial resolution, both laterally and axially, and the depth selectivity are significantly enhanced due to the wide angular spectrum of the speckle wave. We applied our method to image the dynamics of small intracellular particles in live biological cells. With enhanced phase sensitivity and speed, the proposed method will serve as a useful tool to study the dynamics of biological specimens.

Real-time phase-contrast imaging of photothermal treatment of head and neck squamous cell carcinoma: an in vitro study of macrophages as a vector for the delivery of gold nanoshells

Abstract. Photothermal treatment (PTT) using nanoparticles has gained attention as a promising alternative therapy for malignant tumors. One strategy for increasing the selectivity of PTT is the use of macrophages as a cellular vector for delivering nanoparticles. The aim of the present study is to examine the use of macrophages as a cellular vector for efficient PTT and determine the appropriate irradiation power and time of a near-infrared (NIR) laser using real-time phase-contrast imaging. Thermally induced injury and death of cancer cells were found to begin at 44°C to 45°C, which was achieved using the PTT effect with gold nanoshells (NS) and irradiation with a NIR laser at a power of 2 W for 5 min. The peritoneal macrophage efficiently functioned as a cellular vector for the NS, and the cancer cells surrounding the NS-loaded macrophages selectively lost their cellular viability after being irradiated with the NIR laser.

Synthetic aperture microscopy for high resolution imaging through a turbid medium

We report on synthetic aperture microscopy through a highly turbid medium. We first recorded a transmission matrix for the turbid medium with an angular basis of 20,000 complex images covering 0.6 NA. This effectively converts the medium into a lens of the same NA. Distorted images of a target object are then taken at 500 different angles of illumination covering 0.6 NA. For each of the distorted images, the original object image is reconstructed from the transmission matrix by the recently developed turbid lens imaging (TLI) technique. All 500 reconstructed images are synthesized to enhance the NA to 1.2 and thereby generate an object image with twice the enhanced spatial resolution of the individual images. Our method of applying aperture synthesis for TLI makes it possible to enhance the resolving power without increasing the number of transmission matrix elements. This relieves the demand for data acquisition and processing that has impeded the practicality of TLI.

Fractal stochastic modeling of spiking activity in suprachiasmatic nucleus neurons

Individual neurons in the suprachiasmatic nucleus (SCN), the master biological clock in mammals, autonomously produce highly complex patterns of spikes. We have shown that most (~90%) SCN neurons exhibit truly stochastic interspike interval (ISI) patterns. The aim of this study was to understand the stochastic nature of the firing patterns in SCN neurons by analyzing the ISI sequences of 150 SCN neurons in hypothalamic slices. Fractal analysis, using the periodogram, Fano factor, and Allan factor, revealed the presence of a 1/f-type power-law (fractal) behavior in the ISI sequences. This fractal nature was persistent after the application of the GABAA receptor antagonist bicuculline, suggesting that the fractal stochastic activity is an intrinsic property of individual SCN neurons. Based on these physiological findings, we developed a computational model for the stochastic SCN neurons to find that their stochastic spiking activity was best described by a gamma point process whose mean firing rate was modulated by a fractal binomial noise. Taken together, we suggest that SCN neurons generate temporal spiking patterns using the fractal stochastic point process.

Replicating Spots in Reaction-Diffusion Systems

We review the phenomenon of replicating spots in reaction-diffusion systems and discuss the mechanism of replication. This phenomenon was discovered in recent experiments on a ferrocyanide-iodate-sulfite reaction-diffusion system. Patterns form in a thin gel layer that is in contact with a continuously fed stirred reservoir. Patterns of spots are observed to undergo a continuous process of growth and multiplication through cell division and death through overcrowding. A similar phenomenon is also found in numerical simulations in one dimension on a four-species model of the ferrocyanide-iodate-sulfite reaction and in simulations in two dimensions of simpler two-species reaction-diffusion models: Gray–Scott model by J. Pearson and FitzHugh–Nagumo model by A. Hagberg and E. Meron.

In vivo photothermal treatment by the peritumoral injection of macrophages loaded with gold nanoshells.

Photothermal treatment methods have been widely studied for their target specificity and potential for supplementing the limitations of conventional surgical treatments. In this study, we conducted in vivo photothermal treatments using macrophages containing nanoshells as live vectors. We injected macrophages at the peritumoral sites and observed that they had penetrated into the tumor approximately 48 hours after injection. Afterwards, we irradiated with a near-infrared laser for 2 minutes at 1 W/cm(2), causing cancer cell death. Our study identified the optimal conditions of the photothermal treatment and confirmed the feasibility of its use in in vivo treatments.