Publication database
Polar organization of H&E dyes in histology tissue revealed by polarimetric nonlinear microscopy
Structural organization of harmonophores used in hematoxylin (H) and eosin (E) staining is studied with polarimetric multimodal second-harmonic generation (SHG), third-harmonic generation (THG) and multiphoton excitation fluorescence (MPF) microscopy in rat tail tendon histology sections. The polarimetric microscopy imaging reveals that hemalums (complexes of hematoxylin and aluminum) are well aligned with C6h symmetry along the collagen fibers in H-stained tissue, while eosin Y is partially aligned along the fibers in E-stained tissue and also follows organization of C6h symmetry. When both hemalum and eosin are used for H&E staining, the dye molecules interact and align noncentrosymmetrically with C6 symmetry along the collagen fibers, while the stained nuclei appear isotropically organized. The polar alignment of the hemalum and eosin complexes increases the achiral second order susceptibility tensor component ratio

in H&E-stained tissue. The alignment of hemalum and eosin molecules, and their complexes in collagenous tissue, must be considered in nonlinear microscopy and polarimetric analysis of H&E-stained histopathology.
Rayleigh wave induced cavitation bubble structures
A localized energy deposition in a thin layer of liquid between two solid glass plates excites waves in the liquid, solids, and their interfaces. Of particular interest is an elastic surface wave (Rayleigh wave) on the liquid–solid interface that travels faster than the shock wave in the liquid. The surface deformation caused by the Rayleigh wave expands the layer of liquid, thereby locally reducing the pressure below the cavitation threshold. The created tension nucleates many cavitation bubbles, which later collapse due to the passage of the trailing shock wave in the liquid. Interestingly, the bubbles are not arranged homogeneously but on concentric rings centered on the location of the energy deposition. We explain the formation of the concentric rings with the interaction between neighboring bubbles. The fluid–structure interaction is modeled with a coupled finite volume solver that couples a multi-phase compressible fluid region (water and bubble gas) with an elastic solid (glass). We find that the nucleation of bubbles in such a geometry relaxes the tension in their immediate vicinity and thereby suppresses the growth of neighboring bubble nuclei. This idea is confirmed by a Rayleigh–Plesset model of a bubble driven by a far-field pressure obtained from the finite volume simulation. The observed ring patterns are thus the result of the successive activation of statistically distributed nucleation sites into explosively expanding cavitation bubbles in an axisymmetric geometry, whose strong interaction on short distances leads to a hindrance of bubble growth in radially distinct regions.
Amplification of Supersonic Microjets by Resonant Inertial Cavitation-Bubble Pair
We reveal for the first time by experiments that within a narrow parameter regime, two cavitation bubbles with identical energy generated in anti-phase develop a supersonic jet. High-resolution numerical simulation shows a mechanism for jet amplification based on toroidal shock wave and bubble necking interaction. The micro-jet reaches velocities in excess of 1000 m/s. We demonstrate that potential flow approximation established for Worthington jets accurately predicts the evolution of the bubble gas-liquid interfaces.
Fingering of a cavitation bubble in a thin gap: Ejection of the reversed boundary layer into the bulk flow
The flow around a bubble, precipitously expanding in a thin gap between flat walls, was found to have a peculiar feature: distinct fingering occurs at the bubble wall, which was observed through the ultra-high speed optical visualization. The effect is attributed to the reversal of the flow within boundary layers, which provokes the growth of instabilities at the inflection point and, when the surface tension is low enough (the local Weber number is high enough), leads to the fingering. In this paper, we show the high speed recordings of the fingering and model the evolution of the radial velocity to quantitatively confirm feasibility of the proposed instability mechanism.
High-speed ultrasound imaging of bubbly flows and shear waves in soft matter
In this methods paper, we explore the capabilities of high-speed ultrasound imaging (USI) to study fast varying and complex multi-phase structures in liquids and soft materials. Specifically, we assess the advantages and the limitations of this imaging technique through three distinct experiments involving rapid dynamics: the fl ow induced by a liquid jet, the dissolution of sub-micron bubbles in water, and the propagation of shear waves in a soft elastic material. The phenomena were simultaneously characterized using optical microscopy and USI with bubbles as contrast agents. In water, we use compounded high-speed USI for tracking a multi-phase flow produced by a jetting bubble diving into a liquid pool at speeds around 20 m/s. These types of jets are produced by focusing a single laser pulse below the liquid surface. Upon breakup, they create a bubbly fl ow that exhibits high reflectivity to the ultrasound signal, enabling the visualization of the subsequent complex turbulent flow. In a second experiment, we demonstrate the potential of USI for recording the stability and diffusive shrinkage of micro- and nanobubbles in water that could not be optically resolved. Puncturing an elastic material with a liquid jet creates shear waves that can be utilized for elastography measurements. We analysed the shape and speed of shear waves produced by different types of jetting bubbles in industrial gelatin. The wave characteristics were simultaneously determined by implementing particle velocimetry in optical and ultrasound measurements. For the latter, we employed a novel method to create homogeneously distributed micro- and nanobubbles in gelatin by illuminating it with a collimated laser beam.
Robust cavitation-based pumping into a capillary
Cavitation bubbles collapsing near boundaries create liquid flow through their center of mass movement, the formation of liquid jets, and long living vorticities. Here, we demonstrate robust pumping of the liquid with a compact and simple geometry, the open end of a thin-walled circular capillary tube filled with liquid. We study the dynamics of the cavitation bubbles and report on the resultant microjet formation through experiments and simulations. In the experiments, the dynamics of laser-induced cavitation bubbles are captured with high-speed microscopy. Simulations show excellent agreement with the experiments. The jet flow pumps liquid flow toward the capillary opening. The simulation reveals that, in the current study range, both the non-dimensional inner diameter of the capillary and the non-dimensional stand-off distance show influences on the jet width, and only the non-dimensional stand-off distance affects the maximum jet velocities. The results demonstrate that the confinement of the bubble within the capillary alters the anisotropic pressure field around the bubble, leading to a more mild collapse.
Ultrafast transient absorption spectra and kinetics of human blue cone visual pigment at room temperature
The ultrafast photochemical reaction mechanism, transient spectra, and transition kinetics of the human blue cone visual pigment have been recorded at room temperature. Ultrafast time-resolved absorption spectroscopy revealed the progressive formation and decay of several metastable photo-intermediates, corresponding to the Batho to Meta-II photo-intermediates previously observed with bovine rhodopsin and human green cone opsin, on the picosecond to millisecond timescales following pulsed excitation. The experimental data reveal several interesting similarities and differences between the photobleaching sequences of bovine rhodopsin, human green cone opsin, and human blue cone opsin. While Meta-II formation kinetics are comparable between bovine rhodopsin and blue cone opsin, the transition kinetics of earlier photo-intermediates and qualitative characteristics of the Meta-I to Meta-II transition are more similar for blue cone opsin and green cone opsin. Additionally, the blue cone photo-intermediate spectra exhibit a high degree of overlap with uniquely small spectral shifts. The observed variation in Meta-II formation kinetics between rod and cone visual pigments is explained based on key structural differences.
Bulk material influence on the aggressiveness of cavitation – Questioning the microjet impact influence and suggesting a possible way to erosion mitigation
In a study conducted over 10 years ago (Petkovsek and Dular, 2013) [1] we noticed that the thin metal sheet sustains less cavitation damage when it is attached to an acrylic glass (PMMA) than in the case when we attached it to quartz glass (SiO2). The reason for this was not explored at the time.
In the present paper we present a systematic study of single cavitation bubble erosion of a thin aluminum foil, which was attached to either PMMA or SiO2 plate. We show that the damage sustained on the foil attached to PMMA plate is significantly smaller regardless of the bubble collapse distance from the boundary. The result is surprising since one would expect the weak foil to be severely damaged regardless of the material it is attached to.
By femtosecond illumination and high-speed image acquisition we were able to capture the formation and progression of the shock waves, which are emitted at cavitation bubble collapse and observed that they are reflected on SiO2 boundary but that they traverse the PMMA bulk material. We offer an explanation that to achieve less damage the bulk material needs to have acoustic impedance similar to the one of the liquid medium in which cavitation occurs.
Further on, we constructed a simple composite material where PMMA was attached to the SiO2 and showed that we can partially mitigate the damage. This was further confirmed by ultrasonic cavitation erosion tests.
The results also imply that the cavitation damage originates solely from the shock wave, which is emitted at cavitation bubble collapse – consequently putting the idea of microjet impact mechanism under question. Finally, the study offers a new exciting approach to mitigate cavitation erosion by fine tuning the acoustic impedance of the coatings.
Compact, low-cost, and broadband terahertz time-domain spectrometer
Terahertz time-domain spectroscopy (THz-TDS) is a powerful technique that enables the characterization of a large range of bulk materials, devices, and products. Although this technique has been increasingly used in research and industry, the standard THz-TDS configuration relying on the use of a near-infrared (NIR) laser source remains experimentally complex and relatively costly, impeding its availability to those without the expertise to build a high-performance setup based on nonlinear optics or without the financial means to acquire a commercial unit. Broadband THz-TDS systems require an even larger financial investment, primarily because the generation and detection of spectral components exceeding 3 THz typically need an ultrafast NIR source delivering sub-100-fs pulses. Such an ultrafast source can be bulky and cost upwards of $100,000. Here, we present a broadband, compact, and portable THz-TDS system comprising three modules that allow for the implementation of a single low-cost ultrafast laser, hence significantly decreasing the overall cost of the system. In the first module, the output laser pulses are spectrally broadened through nonlinear propagation in a polarization-maintaining optical fiber and then temporally compressed to achieve a higher peak power. The other two modules utilize thick nonlinear crystals with periodically patterned surfaces that diffract NIR pulses and optimize the efficiency of THz generation and detection processes by enabling a noncollinear beam geometry. Phase-matching conditions in the nonlinear crystals are controlled by the period of the gratings to gain access to a large spectral THz bandwidth. The whole system, combining these three modules, provides access to a THz spectrum peaking at 3.5 THz and extending beyond 6 THz with a maximum dynamic range of 50 dB for time-resolved spectroscopy applications. We demonstrate the functionality of this configuration by performing THz spectroscopy measurements of water vapor contained within a closed cell. Our compact system design paves the way towards a high-performance, yet cost-effective, THz-TDS system that can be readily used in academia and industry.
Effect of out of plane orientation on polarization second harmonic generation of single collagen fibrils
Second harmonic generation (SHG) microscopy has emerged as a powerful technique for visualizing collagen organization within tissues. Amongst the many advantages of SHG is its sensitivity to collagen nanoscale organization, and its presumed sensitivity to the relative out of plane polarity of fibrils. Recent results have shown that circular dichroism SHG (CD-SHG), a technique that has been commonly assumed to reveal the relative out of plane polarity of collagen fibrils, is actually insensitive to changes in fibril polarity. However, results from another research group seem to contradict this conclusion. Both previous results have been based on SHG imaging of collagen fibrils within tissues, therefore, to gain a definitive understanding of the sensitivity of SHG to relative out of plane polarity, the results from individual fibrils are desirable. Here we present polarization resolved SHG microscopy (PSHG) data from individual collagen fibrils oriented out of the image plane by buckling on an elastic substrate. We show through correlation with atomic force microscopy measurements that SHG intensity can be used to estimate the out of plane angle of individual fibrils. We then compare the sensitivity of two PSHG techniques, CD-SHG and polarization-in, polarization-out SHG (PIPO-SHG), to the relative out of plane polarity of individual fibrils. We find that for single fibrils CD-SHG is insensitive to relative out of polarity and we also demonstrate the first direct experimental confirmation that PIPO-SHG reveals the relative out of plane polarity of individual collagen fibrils.