Plants exhibit life-long organogenic and histogenic activity in a specialised organ, the shoot apical meristem. Leaves and flowers are formed within the ring-shaped peripheral zone, which surrounds the central zone, the site of the stem cells. We have undertaken a series of high-precision laser ablation and microsurgical tissue removal experiments to test the functions of different parts of the tomato meristem, and to reveal their interactions. Ablation of the central zone led to ectopic expression of the WUSCHELgene at the periphery, followed by the establishment of a new meristem centre. After the ablation of the central zone, organ formation continued without a lag. Thus, the central zone does not participate in organogenesis, except as the ultimate source of founder cells. Microsurgical removal of the external L1 layer induced periclinal cell divisions and terminal differentiation in the subtending layers. In addition, no organs were initiated in areas devoid of L1, demonstrating an important role of the L1 in organogenesis. L1 ablation had only local effects, an observation that is difficult to reconcile with phyllotaxis theories that invoke physical tension operating within the meristem as a whole. Finally, regeneration of L1 cells was never observed after ablation. This shows that while the zones of the meristem show a remarkable capacity to regenerate after interference, elimination of the L1layer is irreparable and causes terminal differentiation.

In medical imaging different techniques have been developed to gain information from inside a tissue. Optoacoustics is a method to generate tomography pictures of tissue using Q-switched laser pulses. Due to thermal and pressure confinement, a short light pulse generates a pressure distribution inside tissue, which mirrors absorbing structures and can be measured outside the tissue. Using a temporal back-projection method, the pressure distribution measured on the tissue surface allows us to gain a tomography picture of the absorbing structures inside tissue. This study presents a novel computational algorithm, which, at least in principle, yields an exact reconstruction of the absorbing structures in three-dimensional space inside the tissue. The reconstruction is based on 2D pressure distributions captured outside at different delay times. The algorithm is tested in a simulation and back-projection of pressure transients of a small absorber and a single point source.

Leaves are arranged according to regular patterns, a phenomenon referred to as phyllotaxis. Important determinants of phyllotaxis are the divergence angle between successive leaves, and the size of the leaves relative to the shoot axis. Young leaf primordia are thought to provide positional information to the meristem, thereby influencing the positioning of new primordia and hence the divergence angle. On the contrary, the meristem signals to the primordia to establish their dorsoventral polarity, which is a prerequisite for the formation of a leaf blade. These concepts originate from classical microsurgical studies carried out between the 1920s and the 1970s. Even though these techniques have been abandoned in favor of genetic analysis, the resulting insights remain a cornerstone of plant developmental biology.Here, we employ new microsurgical techniques to reassess and extend the classical studies on phyllotaxis and leaf polarity. Previous experiments have indicated that the isolation of an incipient primordium by a tangential incision caused a change of divergence angle between the two subsequent primordia, indicating that pre-existing primordia influence further phyllotaxis. Here, we repeat these experiments and compare them with the results of laser ablation of incipient primordia. Furthermore, we explore to what extent the different pre-existing primordia influence the size and position of new organs, and hence phyllotaxis. We propose that the two youngest primordia (P1 and P2) are sufficient for the approximate positioning of the incipient primordium (I1), and therefore for the perpetuation of the generative spiral, whereas the direct contact neighbours of I1 (P2 and P3) control its delimitation and hence its exact size and position. Finally, we report L1-specific cell ablation experiments suggesting that the meristem L1 layer is essential for the dorsoventral patterning of leaf primordia.

Widespread applications of ZnO nanoparticles (NP) in sun-blocking cosmetic products have raised safety concerns related to their potential transdermal penetration and resultant cytotoxicity. Nonlinear optical microscopy provides means for high-contrast imaging of ZnO NPs lending in vitro and in vivo assessment of the nanoparticle uptake in skin, provided their nonlinear optical properties are characterized. We report on this characterization using ZnO NP commercial product, Zinclear, mean-sized 21 nm. Two-photon action cross-section of this bandgap material (E_bg = 3.37 eV, λ_bg = 370 nm) measured by two techniques yielded consistent results of ${\eta }_{ZnO}{\sigma }_{ZnO}^{\left(2ph\right)}$ = 6.2 ± 0.8 μGM at 795 nm, and 32 ± 6 μGM at 770 nm per unit ZnO crystal cell, with the quantum efficiency of ${\eta }_{ZnO}$ = (0.9 ± 0.2) %. In order to demonstrate the quantitative imaging, nonlinear optical microscopy images of the excised human skin topically treated with Zinclear were acquired and processed using ${\sigma }_{ZnO}^{\left(2ph\right)}$ and ${\eta }_{ZnO}$values yielding nanoparticle concentration map in skin. Accumulations of Zinclear ZnO nanoparticles were detected only on the skin surface and in skin folds reaching concentrations of 800 NPs per μm³.