Second Harmonic Generation Signal Intensity Research Articles

Excited States Dynamics at Pentacene/Perfluoropentacene Interfaces: A Femtosecond Time-Resolved Second Harmonic Generation Study.

Understanding the dynamics of excited states after optical excitation at donor-acceptor (D/A) interfaces is of paramount importance for improving the efficiency and performance of optoelectronic devices. Here, we studied the ultrafast excited state dynamics after optical excitation at interfaces between the electron donor (D) pentacene (PEN) and the electron acceptor (A) perfuoropentacene (PFP) as well as within the single compounds (PEN and PFP) using femtosecond (fs) time-resolved second harmonic generation (SHG). In the single compounds singlet fission is observed on a time scale of around 200 fs. In the bilayer systems a huge SHG intensity rise is observed due to the creation of charge transfer states at the interface and accordingly to formation of a local electric field within tens of picoseconds. The local electric field and therefore the SHG signal intensity from the interface of PEN/PFP bilayer is much more intense compared to the PFP/PEN system because the PFP and PEN intermixing at the PEN/PFP interface is higher. Accordingly a population of defect states on a time scale of 55±12 ps has been proposed for PEN/PFP. Our study provides important insights into D/A charge transfer properties, which is needed for the understanding of the interfacial photophysics of pentacene-based organic compounds.

Adsorption Structures Affecting the Electronic Properties and Photoinduced Charge Transfer at Perylene-Based Molecular Interfaces.

Perylene-based organic semiconductors are widely used in organic electronic devices. Here, we studied the ultrafast excited state dynamics after optical excitation at interfaces between the electron donor (D) diindenoperylene (DIP) and the electron acceptor (A) dicyano-perylene-bis(dicarboximide) (PDIR-CN2 ) using femtosecond time-resolved second harmonic generation (SHG) in combination with large scale quantum chemical calculations. Thereby, we varied in bilayer structures of DIP and PDIR-CN2 the interfacial molecular geometry. For an interfacial configuration which contains a edge-on geometry but also additional face-on domains an optically induced charge transfer (CT) is observed, which leads to a pronounced increase of the SHG signal intensity due to electric field induced second harmonic generation. The interfacial CT state decays within 7.5±0.7 ps, while the creation of hot CT states leads to a faster decay (5.3±0.2 ps). For the bilayer structures with mainly edge-on geometries interfacial CT formation is suppressed since π-π overlap perpendicular to the interface is missing. Our combined experimental and theoretical study provides important insights into D/A charge transfer properties, which is needed for the understanding of the interfacial photophysics of these molecules.

Single fibre cytoarchitecture in ventilator-induced diaphragm dysfunction (VIDD) assessed by quantitative morphometry second harmonic generation imaging: Positive effects of BGP-15 chaperone co-inducer and VBP-15 dissociative corticosteroid treatment.

Ventilator-induced diaphragm dysfunction (VIDD) is a common sequela of intensive care unit (ICU) treatment requiring mechanical ventilation (MV) and neuromuscular blockade (NMBA). It is characterised by diaphragm weakness, prolonged respirator weaning and adverse outcomes. Dissociative glucocorticoids (e.g., vamorolone, VBP-15) and chaperone co-inducers (e.g., BGP-15) previously showed positive effects in an ICU-rat model. In limb muscle critical illness myopathy, preferential myosin loss prevails, while myofibrillar protein post-translational modifications are more dominant in VIDD. It is not known whether the marked decline in specific force (force normalised to cross-sectional area) is a pure consequence of altered contractility signaling or whether diaphragm weakness also has a structural correlate through sterical remodeling of myofibrillar cytoarchitecture, how quickly it develops, and to which extent VBP-15 or BGP-15 may specifically recover myofibrillar geometry. To address these questions, we performed label-free multiphoton Second Harmonic Generation (SHG) imaging followed by quantitative morphometry in single diaphragm muscle fibres from healthy rats subjected to five or 10days of MV + NMBA to simulate ICU treatment without underlying confounding pathology (like sepsis). Rats received daily treatment of either Prednisolone, VBP-15, BGP-15 or none. Myosin-II SHG signal intensities, fibre diameters (FD) as well as the parameters of myofibrillar angular parallelism (cosine angle sum, CAS) and in-register of adjacent myofibrils (Vernier density, VD) were computed from SHG images. ICU treatment caused a decline in FD at day 10 as well as a significant decline in CAS and VD from day 5. Vamorolone effectively recovered FD at day 10, while BGP-15 was more effective at day 5. BGP-15 was more effective than VBP-15 in recovering CAS at day 10 although not to control levels. In-register VD levels were restored at day 10 by both compounds. Our study is the first to provide quantitative insights into VIDD-related myofibrillar remodeling unravelled by SHG imaging, suggesting that both VBP-15 and BGP-15 can effectively ameliorate the structure-related dysfunction in VIDD.

Second harmonic generation in air-exposed few-layer black phosphorus

Black phosphorus (BP) shows significant saturable absorption and other unique properties in nonlinear optical effects, and it can be used for nonlinear optical elements and other photonic devices. However, the second harmonic generation (SHG) emission is prohibited due to its inversion symmetry. Here we report emerging SHG emission in BP by exposing BP to oxygen so as to break the inversion symmetry. The intensity of SHG signal shows a dependence on thickness and exposure time. It is interesting to find that the thinner BP flakes have faster SHG intensity change rate. Our results demonstrate that the intensity of SHG signal is attributed to oxidation degree. Air exposure also causes a blue shift of photoluminescence (PL) peaks, which is consistent with our calculation results. Our study demonstrates a feasible approach to modulate the SHG emission of BP, which may provide insights to other inversion symmetric crystals like BP. • We report emerging SHG emission in BP by exposing BP to oxygen so as to break the inversion symmetry. • It is founded that the intensity of SHG signal shows a dependence on thickness and exposure time. • It is founded that air exposure causes a blue shift of PL peaks, which is consistent with our calculation results. The results demonstrate that the intensity of SHG signal is attributed to oxidation degree. • To the best of our knowledge, there is no report on the SHG in BP before. This work demonstrates a feasible approach to modulate the SHG emission of BP, which may provide insights to other inversion symmetric crystals like BP.

Anterior cruciate ligament microfatigue damage detected by collagen autofluorescence in situ

PurposeCertain types of repetitive sub-maximal knee loading cause microfatigue damage in the human anterior cruciate ligament (ACL) that can accumulate to produce macroscopic tissue failure. However, monitoring the progression of that ACL microfatigue damage as a function of loading cycles has not been reported. To explore the fatigue process, a confocal laser endomicroscope (CLEM) was employed to capture sub-micron resolution fluorescence images of the tissue in situ. The goal of this study was to quantify the in situ changes in ACL autofluorescence (AF) signal intensity and collagen microstructure as a function of the number of loading cycles.MethodsThree paired and four single cadaveric knees were subjected to a repeated 4 times bodyweight landing maneuver known to strain the ACL. The paired knees were used to compare the development of ACL microfatigue damage on the loaded knee after 100 consecutive loading cycles, relative to the contralateral unloaded control knee, through second harmonic generation (SHG) and AF imaging using confocal microscopy (CM). The four single knees were used for monitoring progressive ACL microfatigue damage development by AF imaging using CLEM.ResultsThe loaded knees from each pair exhibited a statistically significant increase in AF signal intensity and decrease in SHG signal intensity as compared to the contralateral control knees. Additionally, the anisotropy of the collagen fibers in the loaded knees increased as indicated by the reduced coherency coefficient. Two out of the four single knee ACLs failed during fatigue loading, and they exhibited an order of magnitude higher increase in autofluorescence intensity per loading cycle as compared to the intact knees. Of the three regions of the ACL - proximal, midsubstance and distal - the proximal region of ACL fibers exhibited the highest AF intensity change and anisotropy of fibers.ConclusionsCLEM can capture changes in ACL AF and collagen microstructures in situ during and after microfatigue damage development. Results suggest a large increase in AF may occur in the final few cycles immediately prior to or at failure, representing a greater plastic deformation of the tissue. This reinforces the argument that existing microfatigue damage can accumulate to induce bulk mechanical failure in ACL injuries. The variation in fiber organization changes in the ACL regions with application of load is consistent with the known differences in loading distribution at the ACL femoral enthesis.

Multiphoton microscopy assessment of the structure and variability changes of dermal connective tissue in vulvar lichen sclerosus: A pilot study.

In this article, we offer a novel classification of progressive changes in the connective tissue of dermis in vulvar lichen sclerosus (VLS) relying on quantitative assessment of the second harmonic generation (SHG) signal received from formalin fixed and deparaffinized tissue sections. We formulate criteria for distinguishing four degrees of VLS development: Initial-Mild-Moderate-Severe. Five quantitative characteristics (length and thickness type I Collagen fibers, Mean SHG signal intensity, Skewness and Coherence SHG signal) are used to describe the sequential degradation of connective tissue (changes in the structure, orientation, shape and density of collagen fibers) up to the formation of specific homogeneous masses. Each of the degrees has a characteristic set of quantitatively expressed features. We focus on the identification and description of early, initial changes of the dermis as the least specific. The results obtained by us and the proposed classification of the degrees of the disease can be used to objectify the dynamics of tissue changes during treatment.

Augmentation of surface plasmon-enhanced second harmonic generation from Au nanoprisms on SiO2/Si: interference contribution.

We present an augmentation of Surface Plasmon (SP)-enhanced second harmonic generation (SHG) due to interference field enhancement in Au nanoprisms (AuNPs) on SiO2-coated Si substrates. The SiO2 spacer contributed for the optical interference and increased the coupling efficiency of the pump light with the SP polarization as well as a decoupling efficiency of the SHG waves from nonlinear polarization. The intensity of the SP-enhanced SHG signals increased 4.5-fold with respect to the AuNPs on the bare SiO2 substrate by setting the SiO2 spacer layer to the appropriate thickness. The numerical analysis revealed that the optimal SHG conversion was determined by the balance between the degree of the optical interference at the fundamental and SHG wavelengths.

Second harmonic generation microscopy of otoconia.

The origin of second harmonic generation (SHG) signal in otoconia was investigated. SHG signal intensity from otoconia was compared to pure calcite crystals, given calcite is the primary component of otoconia and is known to emit surface SHG. The SHG intensity from calcite was found to be ∼41× weaker than the SHG intensity from otoconia signifying that the SHG signal from otoconia is likely generated from the organic matrix. Furthermore, the SHG intensity from otoconia increased when treated with a chelating agent known to dissolve calcite which confirms that calcite is not the source of SHG. Additionally, polarization-resolved SHG microscopy imaging revealed that the arrangement of the SHG emitters is radial and can form highly ordered domains.

K2 Na3 [{B6 O10 (OH)}{B3 O4 (OH)3 }] ⋅ H2 O: A Layered Borate Built by Mixed Oxoboron Clusters with Nonlinear-Optical Property.

A new acentric borate, K2 Na3 [{B6 O10 (OH)}-{B3 O4 (OH)3 }] ⋅ H2 O (1) has been made under solvothermal condition. 1 has layered structure made by B6 O13 (OH)-based chains and B3 O5 (OH)3 -bridging clusters. Second-harmonic generation (SHG) measurements reveal that 1 is a phase-matchable nonlinear optical (NLO) material, showing the SHG signal intensity of 1.8 times that of KDP (KH2 PO4 ). Besides, UV-Vis diffuse reflectance spectrum shows that 1 has the short deep UV (DUV) absorption cutoff edge of 198 nm. Thermogravimetric analysis reveals it has good thermal stability. Also 1 represents firstly mixed oxoboron clusters-made 2D layered borate with NLO property.

Late Changes in the Extracellular Matrix of the Bladder after Radiation Therapy for Pelvic Tumors.

Radiation therapy is one of the cardinal approaches in the treatment of malignant tumors of the pelvis. It leads to the development of radiation-induced complications in the normal tissues. Thus, the evaluation of radiation-induced changes in the extracellular matrix of the normal tissue is deemed urgent, since connective tissue stroma degradation plays a crucial role in the development of Grade 3–4 adverse effects (hemorrhage, necrosis, and fistula). Such adverse effects not only drastically reduce the patients’ quality of life but can also become life-threatening. The aim of this study is to quantitatively analyze the bladder collagen state in patients who underwent radiation therapy for cervical and endometrial cancer and in patients with chronic bacterial cystitis and compare them to the normal bladder extracellular matrix. Materials and methods: One hundred and five patients with Grade 2–4 of radiation cystitis, 67 patients with bacterial chronic cystitis, and 20 volunteers without bladder pathology were enrolled. Collagen changes were evaluated depending on its hierarchical level: fibrils and fibers level by atomic force microscopy; fibers and bundles level by two-photon microscopy in the second harmonic generation (SHG) mode; general collagen architectonics by cross-polarization optical coherence tomography (CP OCT). Results: The main sign of the radiation-induced damage of collagen fibrils and fibers was the loss of the ordered “basket-weave” packing and a significant increase in the total area of ruptures deeper than 1 µm compared to the intact sample. The numerical analysis of SHG images detected that a decrease in the SHG signal intensity of collagen is correlated with the increase in the grade of radiation cystitis. The OCT signal brightness in cross-polarization images demonstrated a gradual decrease compared to the intact bladder depending on the grade of the adverse event. Conclusions: The observed correspondence between the extracellular matrix changes at the microscopic level and at the level of the general organ architectonics allows for the consideration of CP OCT as a method of “optical biopsy” in the grading of radiation-induced collagen damage.

Observations of intracellular second-harmonic generation imaging in black phosphorus nanosheets

Functionalized black phosphorus (BP) nanosheets have been considered as promising nanoagents in cancer therapy due to their excellent photothermal conversion efficiency. However, it is still difficult to visually monitor the dynamic localization of BP nanoagents in cancer cells. In this paper, we systematically studied the second-harmonic generation (SHG) signals originating from exfoliated BP nanosheets. Interestingly, under the excitation of a high frequency pulsed laser at 950 nm, the SHG signals of BP nanosheets in vitro are almost undetectable because of their poor stability. However, the intracellular SHG signals from BP nanosheets could be measured by in vivo optical [Formula: see text]imaging due to the efficient enrichment of living HeLa cells. Moreover, the SHG signal intensity from BP nanosheets increases with the prolonged incubation time. It can be expected that the BP nanosheets could be a promising intracellular SHG nanoprobe employed for visually in vivo biomedical imaging in practical cancer photothermal therapy (PIT).

An intrinsic, label-free signal for identifying stem cell-derived cardiomyocyte subtype.

Human-induced pluripotent stem cell (hiPSC)-derived cardiomyocytes have many promising applications, including the regeneration of injured heart muscles, cardiovascular disease modeling, and drug cardiotoxicity screening. Current differentiation protocols yield a heterogeneous cell population that includes pluripotent stem cells and different cardiac subtypes (pacemaking and contractile cells). The ability to purify these cells and obtain well-defined, controlled cell compositions is important for many downstream applications; however, there is currently no established and reliable method to identify hiPSC-derived cardiomyocytes and their subtypes. Here, we demonstrate that second harmonic generation (SHG) signals generated directly from the myosin rod bundles can be a label-free, intrinsic optical marker for identifying hiPSC-derived cardiomyocytes. A direct correlation between SHG signal intensity and cardiac subtype is observed, with pacemaker-like cells typically exhibiting ~70% less signal strength than atrial- and ventricular-like cardiomyocytes. These findings suggest that pacemaker-like cells can be separated from the heterogeneous population by choosing an SHG intensity threshold criteria. This work lays the foundation for developing an SHG-based high-throughput flow sorter for purifying hiPSC-derived cardiomyocytes and their subtypes.

Photo-induced non-linear optical studies on gallium alkali borate glasses doped with TiO2

30 Li2O − (5 − x) Ga2O3 − 65 B2O3 glasses containing small concentrations of TiO2 varying from 0 to 1.0 mol% are prepared by the melt quench technique. The X-ray diffraction results showed the amorphous nature of the sample. Various physical parameters like density, molar volume, Ti4+ ion concentration, molar refraction, polarizability, polaron radius, etc., are evaluated. The differential scanning calorimetry traces of all the samples are recorded and displayed inflection points due to glass transition Tg in the temperature region 446–460 °C. Fourier transform infrared spectra, optical absorption spectra and photoluminescence spectra of all the samples are recorded at room temperature and analyzed. Optical absorption spectra of all these glasses exhibited two clearly resolved absorption due to the excitation of Ti3+ ions. At room temperature, the recorded electron paramagnetic resonance (EPR) spectrum of present glasses exhibited a resonance signal at g ~ 1.911 referred to tetragonaly compressed octahedral sites of Ti3+ ions. Laser-induced second-harmonic generation (SHG) studies are carried out for all the glasses. These results have signified that the glass doped with 0.8 mol% of TiO2 exhibited the highest intensity of SHG signal. These results are further analyzed using the other spectroscopic studies and reasons for the highest SHG efficiency of the glass GT8 are identified and reported.

Quantitative assessment of radiation-induced changes of bladder and rectum collagen structure using optical methods

The objective of the study is the quantitative analysis of the dose-time dependences of changes occurring in collagen of bladder and rectum after gamma-irradiation using optical methods [nonlinear microscopy in a second harmonic generation (SHG) detection regime and cross-polarization optical coherence tomography (CP OCT)]. For quantitative assessment of the collagen structure, regions of interest on the SHG-images of two-dimensional (2-D) distribution of SHG signal intensity of collagen were chosen in the submucosa. The mean SHG signal intensity and its standard deviation were calculated by ImageJ 1.39p (NIH). For quantitative analysis of CP OCT data, an integral depolarization factor (IDF) was calculated. Quantitative calculation of the SHG signal intensity and the IDF can provide additional information about the processes of the collagen radiation-induced degradation and subsequent remodeling. High positive correlation between the mean SHG signal intensity and the mean IDF of bladder and rectum demonstrates that CP OCT can be used as an "optical biopsy" in the grading of collagen radiation damage.

Identification of inactive conformation-selective interleukin-2-inducible T-cell kinase (ITK) inhibitors based on second-harmonic generation.

Many clinically approved protein kinase inhibitors stabilize an inactive conformation of their kinase target. Such inhibitors are generally highly selective compared to active conformation inhibitors, and consequently, general methods to identify inhibitors that stabilize an inactive conformation are much sought after. Here, we have applied a high‐throughput, second‐harmonic generation (SHG)‐based conformational approach to identify small molecule stabilizers of the inactive conformation of interleukin‐2‐inducible T‐cell kinase (ITK). A single‐site cysteine mutant of the ITK kinase domain was created, labeled with an SHG‐active dye, and tethered to a supported lipid bilayer membrane. Fourteen tool compounds, including stabilizers of the inactive and active conformations as well as nonbinders, were first examined for their effect on the conformation of the labeled ITK protein in the SHG assay. As a result, inactive conformation inhibitors were clearly distinguished from active conformation inhibitors by the intensity of SHG signal. Utilizing the SHG assay developed with the tool compounds described above, we identified the mechanism of action of 22 highly selective, inactive conformation inhibitors within a group of 105 small molecule inhibitors previously identified in a high‐throughput biochemical screen. We describe here the first use of SHG for identifying and classifying inhibitors that stabilize an inactive vs. an active conformation of a protein kinase, without the need to determine costructures by X‐ray crystallography. Our results suggest broad applicability to other proteins, particularly with single‐site labels reporting on specific protein movements associated with selectivity.

Effect of composition and temperature on the second harmonic generation in silver phosphate glasses

We herein employ nonlinear laser imaging microscopy to explicitly study the dynamics of second harmonic generation (SHG) in silver iodide phosphate glasses. While glasses of this family have gained extensive scientific attention over the years due to their superior conducting properties, considerably less attention has been paid to their unique nonlinear optical characteristics. In the present study, firstly, it is demonstrated that SHG signal intensity is enhanced upon increasing silver content due to the random formation of silver microstructures within the glass network. Secondly, the SHG temperature dynamics were explored near the glass transition temperature (Tg) regime, where significant glass relaxation phenomena occur. It is found that heating towards the Tg improves the SHG efficiency, whereas above Tg, the capacity of glasses to generate second harmonic radiation is drastically suppressed. The novel findings of this work are considered important in terms of the potential employment of these glasses for the realization of advanced photonic applications like optical-switches and wavelength conversion devices.

Low Temperature Fluorinated Silicon FILMS Synthesis

Second-harmonic generation (SHG), photoluminescence (PL) and Fourier-transformed infrared (FT IR) spectra from silicon films with different average size of nanocrystals were analized to develop new possible material for active channel in nonlinear optical devices. Silicon films were prepared with dominant concentrations of silicon bonding with oxygen or fluorine atoms. Also, the volume fraction of nanocrystals was varied from 30% to 70%. It is seen the spectral peak with energy 3.26 eV is related to defects appeared in interface area silicon-silicon dioxide. For films with small silicon crystals (less than 20 nm) the nonlinear optical response contains two spectral peaks. The second peak is caused by optical response from nanocrystal grain boundary that contains oxygen or fluorine atoms incorporated in silicon as dipoles inside film. The optical nonlinear switch device based on the nonlinear optical response of SiOxmedia inside film was proposed. Also, the silicon film with quartz micro-clusters was investigated as material for making the nonlinear optical transmitter device. The structural properties of silicon films can be easily illustrated by using the films morphology which was determined by using atomic-force microscopy. Figure 1 shows the atomic-force microscopic photos for Sample 1, Sample 2 and Sample3, respectively. It is seen, that the nanocrystal grains are in all types of investigated silicon films. The lateral distribution of nanocrystals with dominant orientation (111) are homogeneous for Sample 1 (see Fig. 1 a)), but for the Sample 2 is unhomogeneous due to appeared conglomerates which are created from the amorphous phase separately (see Fig. 1 b)). The Sample 3 has the triangular shape of nanocrystals (111) which are distributed in submicron crystalline (111) domens. It is clear, that there is great difference in morphological properties of three types of films. The HF etching the silicon surface is efficient because at Td=100oC the surface diffusion and desorption from the surface are low. By adding the SiF4 we change the H2 :SiF4 ratio and some amount of fluorine atoms are included into H2SiF6 assembly creation. Because, the etching process is low at [SiF4] =0.25 sccm and [SiF4] =0.375 sccm. After this values of silicon tetra-fluoride flow rate the H2:SiF4 ratio is changed to destruct intensive creation of molecular assemblies. This stage at [SiF4] =0.5 sccm the HF significant etches the silicon and causes the decreasing the grain sizes. It is assumed that there are two possibilities of fluorine removing: the first is hydrogen incorporation with fluorine, and, the second, the H2SiF6 assemble is creation by 300oC. The SHG is forbidden for centrosymmetric crystal such as bulk silicon because the sum dipole moment is zero, but it is possible due to the surface breaking symmetry and quadruple terms contributions. The opposite situation is for nanostructured oxidized (or fluorinated) silicon films for which the surface area of great quantity of nanocrystals is significant, and the breaking of symmetry is permanent and lateral isotropic. The spectral characteristic of optical nonlinear response from the thin silicon film strictly depends on the spectral properties of polarization and susceptibility tensors χ(2) distortion and χ(2) Si-O.F are the nonlinear susceptibilities produced by nonzero dipole moment caused by shape distortion from the spherical shape of nanocrystals and due to the existing of dangling bonds caused by fluorine or oxygen incorporation in grain boundary region, respectively. By these values of densities the dipole moments of such nanoscaled ellipsoidal dipoles covered by surface charges can be estimated as PSiO =0.04 Debye and PSiH =0.015 Debye. Therefore, the concentration of nanocrystals mainly plays a great role in PL and SHG signal intensity of radiation due to their great surface area covered by silicon bonds with contaminants (atoms of O, H, F) in grain boundary but not as a separate nanoscaled dipoles. By increasing the nanocrystal quantity inside the silicon film the ratio S/V grows significant. The size effect in PL from nanocrystalline silicon film can be explained by means of statistical method. We suggested that the ƒ(x) is distribution function of crystallites in their sizes, θ(x) is a quantum efficiency of nanocrystalls. It is supposed, that the large quantity of nanocrystals is Gauss distributed in their sizes. The Nnc-Si value is proportional to the square under the both functions ƒ(x) and θ(x) near the zero values. Accordingly, the quantity of emitted hydrogenized nanocrystals by band-to-band radiative transitions is important to esimate the PL linear optical response. For nanocrystalline silicon films for Pin àSout optical scheme χ(2) xxx component of susceptibility tensor is detected. By using optical scheme Pin àPout SHG spectra contains isotropic and anisotropic component of susceptibility tensor χ(2) zzz, χ(2) zxx,χ(2) xxz , and χ(2) xxx. Figure 1

Deep‐Ultraviolet Nonlinear Optics in a Borate Framework with 21‐Ring Channels

A new borate LiBa3 (OH)[B9 O16 ][B(OH)4 ], which combines the uniform porosity of open-frameworks with the extraordinary NLO properties of borates, has been obtained under hydrothermal conditions by using mixed lithium and barium ions as templates. The framework displays an acs-type net with large 21-ring channels. The second harmonic generation (SHG) measurement shows that it is a type I phase-matchable material with a strong SHG signal intensity about 3.1 times that of KDP (KH2 PO4 ). UV/Vis-NIR diffuse reflectance analysis indicates that the compound has a wide transparency range with the short-wavelength absorption edge below 200 nm. These characteristics reveal that the compound is a promising deep-UV nonlinear optical material.

Continuous Grading of Early Fibrosis in NAFLD Using Label-Free Imaging: A Proof-of-Concept Study.

Background and AimsEarly detection of fibrosis is important in identifying individuals at risk for advanced liver disease in non-alcoholic fatty liver disease (NAFLD). We tested whether second-harmonic generation (SHG) and coherent anti-Stokes Raman scattering (CARS) microscopy, detecting fibrillar collagen and fat in a label-free manner, might allow automated and sensitive quantification of early fibrosis in NAFLD.MethodsWe analyzed 32 surgical biopsies from patients covering histological fibrosis stages 0–4, using multimodal label-free microscopy. Native samples were visualized by SHG and CARS imaging for detecting fibrillar collagen and fat. Furthermore, we developed a method for quantitative assessment of early fibrosis using automated analysis of SHG signals.ResultsWe found that the SHG mean signal intensity correlated well with fibrosis stage and the mean CARS signal intensity with liver fat. Little overlap in SHG signal intensities between fibrosis stages 0 and 1 was observed. A specific fibrillar SHG signal was detected in the liver parenchyma outside portal areas in all samples histologically classified as having no fibrosis. This signal correlated with immunohistochemical location of fibrillar collagens I and III.ConclusionsThis study demonstrates that label-free SHG imaging detects fibrillar collagen deposition in NAFLD more sensitively than routine histological staging and enables observer-independent quantification of early fibrosis in NAFLD with continuous grading.

The Impact of Collagen Fibril Polarity on Second Harmonic Generation Microscopy

In this work, we report the implementation of interferometric second harmonic generation (SHG) microscopy with femtosecond pulses. As a proof of concept, we imaged the phase distribution of SHG signal from the complex collagen architecture of juvenile equine growth cartilage. The results are analyzed in respect to numerical simulations to extract the relative orientation of collagen fibrils within the tissue. Our results reveal large domains of constant phase together with regions of quasi-random phase, which are correlated to respectively high- and low-intensity regions in the standard SHG images. A comparison with polarization-resolved SHG highlights the crucial role of relative fibril polarity in determining the SHG signal intensity. Indeed, it appears that even a well-organized noncentrosymmetric structure emits low SHG signal intensity if it has no predominant local polarity. This work illustrates how the complex architecture of noncentrosymmetric scatterers at the nanoscale governs the coherent building of SHG signal within the focal volume and is a key advance toward a complete understanding of the structural origin of SHG signals from tissues.