Membrane backtracking at the maximum capacity of nondigestible antigen phagocytosis in macrophages.

The zipper model has been dominantly used to describe the driving mechanism of the engulfment process and its specific identification of antigens during phagocytosis in macrophages. However, the abilities and limitations of the zipper model, capturing the process as an irreversible reaction, have not been examined yet under the critical conditions of engulfment capacity. Here, we demonstrated the phagocytic behavior of macrophages after reaching the maximum engulfment capacity by tracking the progression of their membrane extension during engulfment using IgG-coated nondigestible polystyrene beads and glass microneedles. The results showed that, after macrophages reached their maximum engulfment capacity, they induced membrane backtracking (the reverse phenomenon of engulfment) in both polystyrene beads and glass microneedles, regardless of the difference in the shape of these antigens. We evaluated the correlation of engulfment in simultaneous stimulations of two IgG-coated microneedles and found that each microneedle was regurgitated by the macrophage independently of the advancement or backtracking of membranes on the other microneedle. Moreover, assessing the total engulfment capacity determined by the maximum amount the macrophage was capable of engulfing when imposing each antigen geometry showed that the capacity increased as the attached antigen areas increased. These results indicate that the mechanism of engulfment should imply the following: 1) macrophages have a backtracking function to recover their phagocytic activity after reaching maximal engulfment limit, 2) both phagocytosis and backtracking are local phenomena of the macrophage membrane that operates independently, and 3) the maximum engulfment capacity is determined not only by mere local cell membrane capacity but also by the whole-cell volume increase during simultaneous phagocytosis of multiple antigens by the single macrophages. Thus, the phagocytic activity may entail a hidden backtracking function, adding to the conventionally known irreversible zipper-like ligand-receptor binding mechanism during membrane progression to recover the macrophages that are saturated from engulfing targets beyond their capacity.

On-Chip Free-Flow Measurement Revealed Possible Depletion of Macrophages by Indigestible PM2.5 within a Few Hours by the Fastest Intervals of Serial Phagocytosis.

To understand the influence of indigestible particles like particulate matter 2.5 (PM2.5) on macrophages, we examined the time course of the series phagocytosis of indigestible 2 µm polystyrene spheres (PS). Five kinds of antigens were used as samples for phagocytosis; Zymosan, non-coated 2 µm PS, bovine serum albumin (BSA)-coated PS (BSA-PS), IgG-coated PS (IgG-PS), and IgG-BSA-coated PS (IgG/BSA-PS). To keep the surrounding concentration of antigens against single macrophages constant, antigens flowed at a continuous rate of 0.55 µm/s within a culture dish as a free-flow measurement assay (on-chip free-flow method). The interval of series phagocytosis for IgG/BSA-PS was the shortest among five samples; it was six times faster than Zymosan in terms of engulfment frequency, and up to 50 particles were engulfed within two hours, maintaining constant intervals until reaching the maximum number. The rate of increase in the total number of phagocytozed IgG/BSA-PS over time was constant, at 1.5 particles/min, in series phagocytosis with a 33-cell population, indicating that the phagocytosis rate constant remained constant independent of the number of phagocytoses. Reaction model fitting of the results showed that IgG/BSA-PS had the highest efficiency in terms of the phagocytosis rate constant, 2.3 × 10-2 particles/min, whereas those of IgG-PS, BSA-PS, PS, and Zymosan were 1.4 × 10-2, 1.1 × 10-2, 4.2 × 10-3, and 3.6 × 10-3 particles/min, respectively. One-by-one feeding of IgG/BSA-PS with optical tweezers was examined to confirm the phagocytosis intervals, and we found that the intervals remained constant until several times before the maximum number of antigens for engulfment, also indicating no change in the phagocytosis rate constant regardless of the history of former phagocytosis and phagocytosis number. Simultaneous phagocytosis of two IgG-BSA-decorated microneedle engulfments also showed that the initiation and progress of two simultaneous engulfments on the two different places on a cell were independent and had the same elongation velocity. Therefore, each phagocytosis of indigestible antigens does not affect both in series or in simultaneous subsequent phagocytosis until reaching the maximum capacity of the phagocytosis number. The results suggest (1) no change in the phagocytosis rate constant regardless of the history of phagocytosis numbers and attachment timing and positions, and (2) IgG-BSA decoration of indigestible microparticles in blood accelerates their engulfment faster, resulting in a severe shortage of macrophages within the shortest time.

Simulation of optical radiation force distribution in interference patterns and necessary conditions for chiral structure formation on dielectrics.

A chiral structure is formed by the optical radiation force induced by a circularly polarized light that has spin angular momentum; chiral structures are expected to be used for light control devices and molecular chirality discrimination devices. In this paper, we clarify the relationship between the differences in the distributions of the optical radiation force and the possibility of formation of chiral structures. We first simulate the optical radiation force distribution in the case of a Gaussian beam that successfully forms a chiral structure. Given a vector [Formula: see text] with a centre of the light spot [Formula: see text] and polar coordinates [Formula: see text], and an optical radiation force vector [Formula: see text] at [Formula: see text], the angle [Formula: see text] and [Formula: see text] must be constant with respect to the declination angle [Formula: see text] for a chiral structure to form. These conditions are fulfilled in the case of a 6-beam interference pattern, but not in the case of a 4-beam interference pattern, which is consistent with the result that no chiral structure is formed in the latter case. The equations derived for simulation of optical radiation force distribution can be used for any optical intensity distribution, and will be of great help in the research of any dielectrics deformation.

Laser-Induced Transfer of Noble Metal Nanodots with Femtosecond Laser-Interference Processing.

Noble metal nanodots have been applied to plasmonic devices, catalysts, and highly sensitive detection in bioinstruments. We have been studying the fabrications of them through a laser-induced dot transfer (LIDT) technique, a type of laser-induced forward transfer (LIFT), in which nanodots several hundred nm in diameter are produced via a solid-liquid-solid (SLS) mechanism. In the previous study, an interference laser processing technique was applied to LIDT, and aligned Au nanodots were successfully deposited onto an acceptor substrate in a single shot of femtosecond laser irradiation. In the present experiment, Pt thin film was applied to this technique, and the deposited nanodots were measured by scanning electron microscopy (SEM) and compared with the Au nanodots. A typical nanodot had a roundness fr=0.98 and circularity fcirc=0.90. Compared to the previous experiment using Au thin film, the size distribution was more diffuse, and it was difficult to see the periodic alignment of the nanodots in the parameter range of this experiment. This method is promising as a method for producing large quantities of Pt particles with diameters of several hundred nm.

Petapascal Pressure Driven by Fast Isochoric Heating with a Multipicosecond Intense Laser Pulse.

Fast isochoric laser heating is a scheme to heat matter with a relativistic intensity (>10^{18} W/cm^{2}) laser pulse for producing an ultrahigh-energy-density (UHED) state. We have demonstrated an efficient fast isochoric heating of a compressed dense plasma core with a multipicosecond kilojoule-class petawatt laser and an assistance of externally applied kilotesla magnetic fields for guiding fast electrons to the dense plasma. A UHED state of 2.2 PPa is achieved experimentally with 4.6 kJ of total laser energy that is one order of magnitude lower than the energy used in the conventional implosion scheme. A two-dimensional particle-in-cell simulation confirmed that diffusive heating from a laser-plasma interaction zone to the dense plasma plays an essential role to the efficient creation of the UHED state.

Utilization of the high spatial-frequency component in adaptive beam shaping by using a virtual diagonal phase grating.

A square flattop beam is a fundamental shape that is in high demand in various applications, such as ultra-high-power lasers, uniform surface processing and medical engineering. In this experiment, a new and simple scheme of the adaptive beam shaping system to generate a square flattop shape with high uniformity and edge steepness using virtual diagonal phase grating encoded on a spatial-light modulator and a 4f system is proposed. The grating vector kg is non-parallel to the normal vectors kx and ky of the objective beam profile to be extracted; thus, the residual and extracted components hit separately on the Fourier plane of the 4f system. Consequently, using a spatial-frequency filter passing components parallel to kx and ky, the residual components are blocked by the filter without loss of the high spatial-frequency domain of the extracted component. When the width of the filter was 1.0 mm, the edge of the shaped beam increased in height within 20 µm, which is less than 20% of that obtained with conventional vertical phase grating.

Parallel fabrication of spiral surface structures by interference pattern of circularly polarized beams.

Mass migration of photo-isomeric azo-polymers occurs according to the light intensity gradient, and the morphological surface structure can be fabricated by the artificial distribution of light by applying the interference properties of coherent laser light. Recently, the optical radiation force has played an important role in the morphology for dielectric targets, and chiral structures have been fabricated according to the spirally gathering force distribution that arises due to the electric susceptibility. On the contrary, interference laser processing has been applied to process the surface or interior of the material, and nano- or micro-structures in the lattice have been fabricated in a single exposure to the interference pattern. The unit structures are mostly axisymmetric nanowhiskers, nanodrops and nanobumps, among others. In this experiment, interference laser processing of an azo-polymer dielectric target using a circularly polarised continuous-wave (CW) laser was examined, and a spiral structure was successfully fabricated. From the viewpoint of laser processing method, an optical spiral radiation force was introduced in interference laser processing for the first time.

Magnetized fast isochoric laser heating for efficient creation of ultra-high-energy-density states.

Fast isochoric heating of a pre-compressed plasma core with a high-intensity short-pulse laser is an attractive and alternative approach to create ultra-high-energy-density states like those found in inertial confinement fusion (ICF) ignition sparks. Laser-produced relativistic electron beam (REB) deposits a part of kinetic energy in the core, and then the heated region becomes the hot spark to trigger the ignition. However, due to the inherent large angular spread of the produced REB, only a small portion of the REB collides with the core. Here, we demonstrate a factor-of-two enhancement of laser-to-core energy coupling with the magnetized fast isochoric heating. The method employs a magnetic field of hundreds of Tesla that is applied to the transport region from the REB generation zone to the core which results in guiding the REB along the magnetic field lines to the core. This scheme may provide more efficient energy coupling compared to the conventional ICF scheme.

Local Melting of Gold Thin Films by Femtosecond Laser-Interference Processing to Generate Nanoparticles on a Source Target.

Shape- and size-controlled metallic nanoparticles are very important due to their wide applicability. Such particles have been fabricated by chemosynthesis, chemical-vapor deposition, and laser processing. Pulsed-laser deposition and laser-induced dot transfer use ejections of molten layers and solid-liquid-solid processes to fabricate nanoparticles with a radius of some tens to hundreds of nm. In these processes, the nanoparticles are collected on an acceptor substrate. In the present experiment, we used laser-interference processing of gold thin films, which deposited nanoparticles directly on the source thin film with a yield ratio. A typical nanoparticle had roundness fr=0.99 and circularity fcirc=0.869, and the radius was controllable between 69 and 188 nm. The smallest radius was 82 nm on average, and the smallest standard deviation was 3 nm. The simplicity, high yield, and ideal features of the nanoparticles produced by this method will broaden the range of applications of nanoparticles in fields such as plasmonics.

Heating efficiency evaluation with mimicking plasma conditions of integrated fast-ignition experiment.

A series of experiments were carried out to evaluate the energy-coupling efficiency from heating laser to a fuel core in the fast-ignition scheme of laser-driven inertial confinement fusion. Although the efficiency is determined by a wide variety of complex physics, from intense laser plasma interactions to the properties of high-energy density plasmas and the transport of relativistic electron beams (REB), here we simplify the physics by breaking down the efficiency into three measurable parameters: (i) energy conversion ratio from laser to REB, (ii) probability of collision between the REB and the fusion fuel core, and (iii) fraction of energy deposited in the fuel core from the REB. These three parameters were measured with the newly developed experimental platform designed for mimicking the plasma conditions of a realistic integrated fast-ignition experiment. The experimental results indicate that the high-energy tail of REB must be suppressed to heat the fuel core efficiently.

Design of interference using coherent beams configured as a six-sided pyramid.

Various interference pattern formations have been investigated using six countering laser beams that form a six-sided pyramid. Phase shift and amplitude variations among the interfering beams are very useful for designing interference patterns summarized here. The interference patterns are categorized into 32 unit figures, and structural changes with different thresholds are investigated in detail.