Ultrafast laser ablation of 10-nm self-supporting membranes by two-beam interference processing.

Ultrafast laser ablation was applied to process 10-nm self-supporting membranes. The membranes were processed over tens of square micrometers by single-shot irradiation of two visible laser pulses, followed by the realization of periodic sub-microstructures. The fabricated geometry is dependent on the intensity distribution of the superposed input pulses, providing flexibility and facilitating practical micro- and nanoengineering. Ease of designing the processing parameters and speed of processing are the significant advantages of this method compared to focused ion beam (FIB) milling.

Creating electron phase holograms using femtosecond laser interference processing.

Recently, electron beams with structured phase fronts, such as electron vortex beams, have attracted considerable interest. Herein, we present a novel method of fabricating electron phase holograms using a femtosecond laser interference processing. A 35-nm-thick silicon membrane, corresponding to a phase shift of π for 200-keV electrons, was processed using single-shot laser irradiation, whereas processing such thin membranes with a focused ion beam milling technique would be very difficult. This rapid and efficient technique is expected to produce phase diffraction elements for practical applications in a wide range of electron optics fields.

Odorant concentration differentiator for intermittent olfactory signals.

Animals need to discriminate differences in spatiotemporally distributed sensory signals in terms of quality as well as quantity for generating adaptive behavior. Olfactory signals characterized by odor identity and concentration are intermittently distributed in the environment. From these intervals of stimulation, animals process odorant concentration to localize partners or food sources. Although concentration-response characteristics in olfactory neurons have traditionally been investigated using single stimulus pulses, their behavior under intermittent stimulus regimens remains largely elusive. Using the silkmoth (Bombyx mori) pheromone processing system, a simple and behaviorally well-defined model for olfaction, we investigated the neuronal representation of odorant concentration upon intermittent stimulation in the naturally occurring range. To the first stimulus in a series, the responses of antennal lobe (AL) projection neurons (PNs) showed a concentration dependence as previously shown in many olfactory systems. However, PN response amplitudes dynamically changed upon exposure to intermittent stimuli of the same odorant concentration and settled to a constant, largely concentration-independent level. As a result, PN responses emphasized odorant concentration changes rather than encoding absolute concentration in pulse trains of stimuli. Olfactory receptor neurons did not contribute to this response transformation which was due to long-lasting inhibition affecting PNs in the AL. Simulations confirmed that inhibition also provides advantages when stimuli have naturalistic properties. The primary olfactory center thus functions as an odorant concentration differentiator to efficiently detect concentration changes, thereby improving odorant source orientation over a wide concentration range.

Concentric zones for pheromone components in the mushroom body calyx of the moth brain.

The spatial distribution of input and output neurons in the mushroom body (MB) calyx was investigated in the silkmoth Bombyx mori. In Lepidoptera, the brain has a specialized system for processing sex pheromones. How individual pheromone components are represented in the MB has not yet been elucidated. Toward this end, we first compared the distribution of the presynaptic boutons of antennal lobe projection neurons (PNs), which transfer odor information from the antennal lobe to the MB calyx. The axons of PNs that innervate pheromonal glomeruli were confined to a relatively small area within the calyx. In contrast, the axons of PNs that innervate nonpheromonal glomeruli were more widely distributed. PN axons for the minor pheromone component covered a larger area than those for the major pheromone component and partially overlapped with those innervating nonpheromonal glomeruli, suggesting the integration of the minor pheromone component with plant odors. Overall, we found that PN axons innervating pheromonal and nonpheromonal glomeruli were organized into concentric zones. We then analyzed the dendritic fields of Kenyon cells (KCs), which receive inputs from PNs. Despite the strong regional localization of axons of different PN classes, the dendrites of KCs were less well classified. Finally, we estimated the connectivity between PNs and KCs and suggest that the dendritic field may be organized to receive different amounts of pheromonal and nonpheromonal inputs. PNs for multiple pheromone components and plant odors enter the calyx in a concentric fashion, and they are read out by the elaborate dendritic field of KCs.

Constancy and variability of glomerular organization in the antennal lobe of the silkmoth.

We investigated the anatomical organization of glomeruli in the antennal lobes (ALs) of male silkmoths. We reconstructed 10 different ALs and established an identification procedure for individual glomeruli by using size, shape, and position relative to anatomical landmarks. Quantitative analysis of these morphological characteristics supported the validity of our identification strategy. The glomerular organization of the ALs was roughly conserved between different ALs. However, we found individual variations that were reproducibly observed. The combination of a digital atlas with other experimental techniques, such as electrophysiology, optical imaging, and genetics, should facilitate a more in-depth analysis of sensory information processing in silkmoth ALs.

Modular subdivision of mushroom bodies by Kenyon cells in the silkmoth.

In insects, olfactory information in the glomeruli of the antennal lobe, the first olfactory center, is transmitted to the lateral protocerebrum and the calyx of the mushroom body via projection neurons. In male silkmoths (Bombyx mori), arborization patterns in the calyx differ markedly between projection neurons that respond to sex pheromones and those that respond to general odors. However, little is known about the organization of the mushroom body's intrinsic neurons, called Kenyon cells (KCs), which receive the inputs from the projection neurons. We investigated the silkmoth mushroom body and identified four parallel subdivisions in the lobes and pedunculus by immunolabeling with antibodies against the catalytic subunit of protein kinase A in Drosophila melanogaster (DC0) and the neuromodulatory peptide FMRFamide. To further understand the detailed organization of the mushroom body, we performed extensive labeling of individual KCs. We identified four morphological types whose axonal projections corresponded to the subdivisions in the lobes, and found that each type of KC had a characteristic dendritic morphology in the calyx. These results show a correlation between the axonal projections of KCs in the lobes and dendritic morphology in the calyx, and indicate different functional roles for the subdivisions.