Strong Second Harmonic Generation Signal Research Articles

A Universal Strategy for Synthesis of 2D Ternary Transition Metal Phosphorous Chalcogenides.

The 2D ternary transition metal phosphorous chalcogenides (TMPCs) have attracted extensive research interest due to their widely tunable band gap, rich electronic properties, inherent magnetic and ferroelectric properties. However, the synthesis of TMPCs via chemical vapor deposition (CVD) is still challenging since it is difficult to control reactions among multi-precursors. Here, a subtractive element growth mechanism is proposed to controllably synthesize the TMPCs. Based on the growth mechanism, the TMPCs including FePS3 , FePSe3 , MnPS3 , MnPSe3 , CdPS3 , CdPSe3 , In2 P3 S9 , and SnPS3 are achieved successfully and further confirmed by Raman, second-harmonic generation (SHG), and scanning transmission electron microscopy (STEM). The typical TMPCs-SnPS3 shows a strong SHG signal at 1064nm, with an effective nonlinear susceptibility χ(2) of 8.41×10-11 mV-1 , which is about 8 times of that in MoS2 . And the photodetector based on CdPSe3 exhibits superior detection performances with responsivity of 582mAW-1 , high detectivity of 3.19×1011 Jones, and fast rise time of 611µs, which is better than most previously reported TMPCs-based photodetectors. These results demonstrate the high quality of TMPCs and promote the exploration of the optical properties of 2D TMPCs for their applications in optoelectronics.

#3197 A NOVEL INVESTIGATION OF AVF WITH A SECOND HARMONIC GENERATION MICROSCOPY IN PATIENTS WITH END STAGE RENAL DISEASE

Abstract Background and Aims The population of patients with end-stage renal disease is rapidly growing and hemodialysis (HD) remains the most common treatment option. In clinical practice, the formation of arteriovenous fistulas (AVFs) is considered a priority in order to ensure optimal vascular access for HD patients. One of the complications in the formation of AVF is the hyperplasia of the neointimal layer of the vein, which leads to the failure of the fistula. The impact of the arrangement of collagen fibers in the veins of AVFs is little studied and opens wide opportunities for investigations. The structural tissue can be imaged with polarimetric nonlinear microscopy. Collagen fibers have a non-centrosymmetric structure, so they can be effectively imaged by second harmonic generation (SHG) microscopy [1]. The polarimetric nonlinear microscopy, exploiting the multidimentional tensor nature of light-matter interactions, is emerging as a valuable research tool in biomedical investigations and applications to digital pathology, and is only now becoming technically feasible for potential clinical translation [2,3]. This proposal represents a novel application of multimodal polarimetric nonlinear microscopy in histopathology, and biomedical imaging. The aim of this study is to determine the influence of the arrangement of collagen fibers in veins on the formation efficiency of AVFs by using the innovative method of nonlinear SHG microscopy. Method In this prospective study, we first used a novel SHG microscopy method to investigate AVFs venous intima hyperplasia. V. Cephalica segment is being obtained during arteriovenous fistula formation surgery from patients with end stage renal disease and after the removal of aneurysmatic or fibrotic AVF. Cross-sections of venous specimens are being fixed in 10% buffered formalin and stained with hematoxylin and eosin (H&E). Further, sample investigation is being performed in Laser Research Centre, Vilnius University using a home-built multiphoton laser-scanning microscope. The investigation is being performed with the permission of Lithuanian Biomedical Research Ethics Committee. Results We determined the arrangement of collagen fibers in veins before AVF formation and after the removal of aneurysmatic AVFs. It was observed that tunica intima of V. Cephalica is dominated both by endothelial cells and connective tissue made up of high-density collagen fibers. Whereas tunica adventitia consists mainly of collagen fibers. Collagen in tunica intima and adventitia are found to be highly noncentrosymmetric. Thus a strong SHG signal, resulting in high-contrast structural visualization of the V. Cephalica segment is generated. Conclusion Our initial investigations of AVF show clearly visible collagen fibers. The future work will focus on the structural changes of collagen fibers in the vein intimal hyperplasia. Further investigations need to be done to create artificial intelligence-based identification of new predictive collagen markers.

Large Spontaneous Polarization Ferroelectric Property, Switchable Second-Harmonic Generation Responses, and Magnetism in an Fe-Based Compound.

Since the switchable spontaneous polarization of ferroelectric materials endows it with many useful properties such as a large pyroelectric coefficient, switchable spontaneous polarization, and semiconductor, it has a wide range of application prospects, and the research of high-performance molecular ferroelectric materials has become a hot spot. We obtained a 0D organic-inorganic hybrid ferroelectric [(CH3)3NCH2CH2CH3]2FeCl4 (1) with well-defined ferroelectric domains and excellent domain inversion and exhibited a relatively large spontaneous polarization (Ps = 9 μC/m-2) and a Curie temperature (Tc) of 394 K. Furthermore, compound 1 belongs to the non-centrosymmetrical space group Cmc21 and has a strong second-harmonic generation signal. Interestingly, we also performed magnetic tests on 1, which confirmed that it is a magnetic material. This work provides clues for exploring the application of high-performance molecular ferroelectric materials in future multifunctional smart devices.

Noncentrosymmetric Lanthanide-Based MOF Materials Exhibiting Strong SHG Activity and NIR Luminescence of Er3+: Application in Nonlinear Optical Thermometry

Optically active luminescent materials based on lanthanide ions attract significant attention due to their unique spectroscopic properties, nonlinear optical activity, and the possibility of application as contactless sensors. Lanthanide metal-organic frameworks (Ln-MOFs) that exhibit strong second-harmonic generation (SHG) and are optically active in the NIR region are unexpectedly underrepresented. Moreover, such Ln-MOFs require ligands that are chiral and/or need multistep synthetic procedures. Here, we show that the NIR pulsed laser irradiation of the noncentrosymmetric, isostructural Ln-MOF materials (MOF-Er3+ (1) and codoped MOF-Yb3+/Er3+ (2)) that are constructed from simple, achiral organic substrates in a one-step procedure results in strong and tunable SHG activity. The SHG signals could be easily collected, exciting the materials in a broad NIR spectral range, from ≈800 to 1500 nm, resulting in the intense color of emission, observed in the entire visible spectral region. Moreover, upon excitation in the range of ≈900 to 1025 nm, the materials also exhibit the NIR luminescence of Er3+ ions, centered at ≈1550 nm. The use of a 975 nm pulse excitation allows simultaneous observations of the conventional NIR emission of Er3+ and the SHG signal, altogether tuned by the composition of the Ln-MOF materials. Taking the benefits of different thermal responses of the mentioned effects, we have developed a nonlinear optical thermometer based on lanthanide-MOF materials. In this system, the SHG signal decreases with temperature, whereas the NIR emission band of Er3+ slightly broadens, allowing ratiometric (Er3+ NIR 1550 nm/SHG 488 nm) temperature monitoring. Our study provides a groundwork for the rational design of readily available and self-monitoring NLO-active Ln-MOFs with the desired optical and electronic properties.

Revealing natural fluorescence in transparent insect wings by linear and nonlinear optical techniques

For most natural organisms, the physical, chemical and biological aspects of fluorescence emission are poorly understood. For example, to the best of our knowledge, fluorescence from the transparent wings of any of the 3000 known species of cicadas has never been reported in the literature. These wings are known to exhibit anti-reflective properties arising from quasi-periodic arrays of nipples. Our study, using linear and nonlinear optical techniques, including spectrofluorimetry, two-photon fluorescence spectroscopy and Second Harmonic Generation (SHG), reveals the fluorescence properties in the wings the grey and the common cicadas (Cicada orni and Lyristes (Tibicen) plebejus, respectively), as well as the broad-bordered bee hawk-moth (Hemaris fuciformis). The study suggests that fluorescence would be more widespread in transparent insect wings than what was previously believed. Comparing this result to the fluorescence emission from the wings of the Bornean damselfly (Vestalis amabilis), we inferred that this emission probably arises from resilin, a protein reported to enhance wing flexibility. Moreover, the nonlinear optical investigation of the insects’ wings provided further insight into wing structure, indicating that multiphoton techniques add valuable information for the analysis of insect integuments. The strong SHG signal detected from the wing veins implies that these veins are materially organised in a non-centrosymmetric and hence non-random fashion.

Label-free second harmonic generation imaging of cerebral vascular wall in local ischemia mouse model in vivo

Second harmonic generation (SHG) imaging is label-free and non-invasive, and it has been extensively applied in multiple biological and medical studies, but not in the brain in vivo. In this study, we modified classical two photon excited fluorescence (TPEF) system to perform in vivo simultaneous TPEF and SHG imaging in the local ischemia mouse model. In cerebral vascular walls, we found strong SHG signal, which co-localized with collagen. In the continuous 2 days’ in vivo imaging, this SHG signal remained stable in the local ischemic blood vessel in the initial 4 h, then its signal abruptly increased and got spatially thickened 5 h after thrombosis, and this tendency continued in the following 48 h. This study provides direct and precise timeline of rapid collagen change in cerebral vascular walls in vivo, and reveals the subtle but significant temporal-spatial dynamics of this structural signal during local ischemia. Thus, this cerebral in vivo SHG imaging provides a powerful tool to identify the early and subtle pathological change of collagen around clinical key therapeutic time window.

Strong near band-edge excited second-harmonic generation from multilayer 2H Tin diselenide

We report strong second-harmonic generation (SHG) from 2H polytype of multilayer Tin diselenide (SnSe2) for fundamental excitation close to the indirect band-edge in the absence of excitonic resonances. Comparison of SHG and Raman spectra from exfoliated SnSe2 flakes of different polytypes shows strong (negligible) SHG and Raman Eg mode at 109 cm−1 (119 cm−1), consistent with 2H (1T) polytypes. The difference between the A1g–Eg Raman peak positions is found to exhibit significant thickness dependent for the 1T form, which is found to be absent for the 2H form. The observed thickness dependence of SHG with rapid oscillations in signal strength for small changes in flake thickness are in good agreement with a nonlinear wave propagation model considering nonlinear polarization with alternating sign from each monolayer. The nonlinear optical susceptibility extracted from SHG signal comparison with standard quartz samples for 1040 nm excitation is found to be more than 4-times higher than that at 1550 nm. This enhanced nonlinear response at 1040 nm is attributed to the enhanced nonlinear optical response for fundamental excitation close to the indirect band-edge. We also study SHG from heterostructures of monolayer MoS2/multilayer SnSe2 which allows us to unambiguously compare the nonlinear optical response of SnSe2 with MoS2. We find the SHG signal and any interference effect in the overlap region to be dominated by the SnSe2 layer for the excitation wavelengths considered. The comparison of SHG from SnSe2 and MoS2 underscores that the choice of the 2D material for a particular nonlinear optical application is contextual on the wavelength range of interest and its optical properties at those wavelengths. The present works further highlights the usefulness of near band-edge enhancement of nonlinear processes in emerging 2D materials towards realizing useful nanophotonic devices.

Second harmonic generation in two-dimensional transition metal dichalcogenides with growth and post-synthesis defects

Strong second harmonic generation (SHG) in monolayer transition metal dichalcogenides demonstrates great promise for nonlinear photonic applications. However, to utilize these materials in photonic applications, it is imperative to investigate the robustness of the nonlinear optical properties against various defects introduced during or post synthesis. Here we study the SHG in hexagonal monolayer WS2 with synthesis defects, in triangular monolayer WS2 with focused ion beam induced structural defects, and in WS2 under ambient and aqueous environmental conditions. We find that WS2 hexagonal monolayers exhibit significantly more uniform SHG intensity across the sample with a coefficient of variation, than the varying photoluminescence (PL) intensity (). We also show that introducing structural defects in WS2 monolayers leads to diminishing, but not vanishing SHG. To further explore the robustness of SHG in comparison with PL, we show that aged WS2 monolayers have greatly varying PL across different samples (), while exhibiting much more uniform SHG (). Lastly, we find that WS2 monolayers submerged in deionized water shows similar intensity in SHG signal. To understand these experimental results, density functional theory calculations have been performed. Our analysis suggests that potential defects and water introduced to the WS2 monolayers produce relatively little change in the second-order nonlinear susceptibility, whereas nonradiative recombination caused by these defects can significantly affect the PL. These results show a good qualitative agreement with the experimental results.

Near-Unity Polarization of Valley-Dependent Second-Harmonic Generation in Stacked TMDC Layers and Heterostructures at Room Temperature.

With unique valley-dependent optical and optoelectronic properties, 2D transition metal dichalcogenides (2D TMDCs) are promising materials for valleytronics. Second-harmonic generation (SHG) in 2D TMDCs monolayers has shown valley-dependent optical selection rules. However, SHG in monolayer TMDCs is generally weak; it is important to obtain materials with both strong SHG signals and a large degree of polarization. In the work, a variety of inversion-symmetry-breaking (3R-like phase) TMDCs (WSe2 , WS2 , MoS2 ) atomic layers, spiral structures, and heterostructures are prepared, and their SHG polarization is studied. Through circular-polarization-resolved SHG experiments, it is demonstrated that the SHG intensity is enhanced in thicker samples by breaking inversion symmetry while maintaining the degree of polarization close to unity at room temperature. By studying TMDCs with different twist angles and the spiral structures, it is found that there is no significant effect of multilayer interlayer interaction on valley-dependent SHG. The realization of strong SHG with high degree of polarization may pave the way toward a new platform for nonlinear optical valleytronics devices based on 2D semiconductors.

Enhancement of Second-Order Optical Nonlinearity in a Lutetium Selenite by Monodentate Anion Partial Substitution

The systematic modification of the molecular structure of nonlinear optical (NLO)-active materials is a very attractive approach to the improvement of NLO performance. In this study, one lutetium selenite compound Lu(SeO3)(HSeO3)(H2O)·(H2O) (1) was synthesized by hydrothermal means, while a further two lutetium selenites Lu3F(SeO3)4 (2) and Lu(SeO3)(NO3)(H2O) (3) were successfully synthesized by anion partial substitution (APS) of the parent structure 1 under hydrothermal conditions. Specifically, substitution of the [HSeO3]– anions in the noncentrosymmetric 1 by F– or [NO3]− anions with differing denticity leads to the formation of polar 2 and centrosymmetric 3. Our study reveals that the denticity of the secondary functional anions has a significant influence on the coordination environments of the rare-earth-metal cation Lu3+ and consequently the molecular configuration and NLO performance of the resultant multidimensional selenites. In contrast to 1, which displays a weak second-harmonic generation (SHG) response (0.1 × KH2PO4 (KDP)), 2 exhibits a greatly improved NLO performance, including a strong SHG signal (2.5 × KDP, the highest value among rare-earth-metal-based NLO selenites), a wide band gap (3.57 eV) and optical transparency window (0.35–10.3 μm), high thermal stability (∼550 °C), and a large laser damage threshold (36 × AgGaS2). These results suggest that 2, as the first example of a fluorinated lutetium selenite, is a strong NLO candidate crystal spanning a region from the near-ultraviolet to the mid-infrared. These APS studies highlight a new feasible approach toward high-performance NLO crystals.

Mechanism of Extreme Optical Nonlinearities in Spiral WS2 above the Bandgap.

Layered two-dimensional transition-metal dichalcogenides (2D-TMDs) are promising building blocks for ultracompact optoelectronic applications. Recently, a strong second harmonic generation (SHG) was observed in spiral stacked TMD nanostructures which was explained by its low crystal symmetry. However, the relationship between the efficiency of SHG signals and the electronic band structure remains unclear. Here, we show that the SHG signal in spiral WS2 nanostructures is strongly enhanced (∼100 fold increase) not only when the second harmonic signal is in resonance with the exciton states but also when the excitation energy is slightly above the electronic band gap, which we attribute to a large interband Berry connection associated with certain optical transitions in spiral WS2. The giant SHG enhancement observed and explained in this study could promote the understanding and utility of TMDs as highly efficient nonlinear optical materials and potentially lead to a new pathway to fabricate more efficient optical energy conversion devices.

Broadband mid-infrared second harmonic generation using epitaxial polydomain barium titanate thin films

The mid-infrared (mid-IR) second-order optical nonlinearity of the barium titanate (BTO) thin films was characterized by second harmonic generation (SHG). The epitaxial BTO thin films were grown on strontium titanate substrates by pulsed-laser deposition. From the azimuthal-dependent polarized SHG measurements, the tensorial optical nonlinear coefficients, dij, and ferroelectric domain fraction ratio, δAY/δAz, were resolved. Strong SHG signals were obtained at the pumping laser wavelength λ between 3.0 and 3.6 μm. The SHG intensity was linearly dependent upon the square of the pumping laser power. The broadband mid-IR optical nonlinearity enables BTO thin films for applications in chip-scale quantum optics and nonlinear integrated photonic circuits.

Second-harmonic generation in single-layer monochalcogenides: A response from first-principles real-time simulations

Second Harmonic Generation (SHG) of single-layer monochalcogenides, such as GaSe and InSe, has been recently reported [2D Mater. 5 (2018) 025019; J. Am. Chem. Soc. 2015, 137, 79947997] to be extremely strong with respect to bulk and multilayer forms. To clarify the origin of this strong SHG signal, we perform first-principles real-time simulations of linear and non-linear optical properties of these two-dimensional semiconducting materials. The simulations, based on ab-initio many-body theory, accurately treat the electron-hole correlation and capture excitonic effects that are deemed important to correctly predict the optical properties of such systems. We find indeed that, as observed for other 2D systems, the SHG intensity is redistributed at excitonic resonances. The obtained theoretical SHG intensity is an order of magnitude smaller than that reported at the experimental level. This result is in substantial agreement with previously published simulations which neglected the electron-hole correlation, demonstrating that many-body interactions are not at the origin of the strong SHG measured. We then show that the experimental data can be reconciled with the theoretical prediction when a single layer model, rather than a bulk one, is used to extract the SHG coefficient from the experimental data.

Chiral to Chiral Phase Transition in a Novel Hydrogen Bond Type Molecular Ferroelectric

AbstractAlthough great progress has been made for H‐bonds type molecular ferroelectrics, the systems accompanying with chiral to chiral phase transitions are still very spare. Here we display a new hydrogen bond type molecular ferroelectric, the salt of 1‐(1‐naphthyl) ethylammonium‐nopinate, 1, which undergoes a paraelectric–ferroelectric phase transition around 202 K from one chiral point group (P212121) to another chiral point group (P21). The electric hysteresis loop under 100 K directly indicates it is a potential ferroelectric material with maximum spontaneous polarization (Ps) 2.3 uC/cm2. X‐ray single crystal diffraction analysis shows that the significant hydrogen bond change along the b‐axis direction results net dipole moment and leads to the spontaneous polarization. The exceptional dielectric response in the vicinity of 202 K exhibits remarkable dielectric anomalies along the b‐axis direction than the other two directions. The strong SHG (second harmonic generation) signal (‐1.31) without obvious changes dependence on temperature further confirm the chiral to chiral structural phase transition.

Strong SHG Response via High Orientation of Tetrahedral Functional Motifs in Polyselenide A2Ge4Se10 (A = Rb, Cs)

AbstractNonlinear optical (NLO) materials with strong second harmonic generation (SHG) responses are technologically and scientifically important for tunable lasers. This work puts forward a new strategy for designing promising infrared nonlinear optical (IR‐NLO) materials with strong SHG responses by strengthening both the static and induced contributions via the high orientation of NLO functional motifs. Two new polyselenides Rb2Ge4Se10 and Cs2Ge4Se10 are studied to verify the strategy, they have 2D infinite [Ge4Se10]2− layers comprising highly oriented distorted GeSe4 tetrahedra. Their large SHG signals (8.0 and 8.5 times that of commercial AgGaS2) can be ascribed to both the static contribution enhancement from the high orientation of the local dipole moment of GeSe4 tetrahedra and the induced contribution enhancement from terminal and SeSe bonded Se atoms. The results confirm the effectiveness of the proposed approaches, shedding light on the exploration of practical IR‐NLO materials with maximized performance.

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.

Single potassium niobate nano/microsized particles as local mechano-optical Brownian probes.

Perovskite alkaline niobates, due to their strong nonlinear optical properties, including birefringence and the capability to produce second-harmonic generation (SHG) signals, attract a lot of attention as potential candidates for applications as local nano/microsized mechano-optical probes. Here, we report on an implementation of photonic force microscopy (PFM) to explore the Brownian motion and optical trappability of monocrystalline potassium niobate (KNbO3) nano/microsized particles having sizes within the range of 50 to 750 nm. In particular, we exploit the anisotropic translational diffusive regime of the Brownian motion to quantify thermal fluctuations and optical forces of singly-trapped KNbO3 particles within the optical trapping volume of a PFM microscope. We also show that, under near-infrared (NIR) excitation of the highly focused laser beam of the PFM microscope, a single optically-trapped KNbO3 particle reveals a strong SHG signal manifested by a narrow peak (λ(em) = 532 nm) at half the excitation wavelength (λ(ex) = 1064 nm). Moreover, we demonstrate that the thus induced SHG emission can be used as a local light source that is capable of optically exciting molecules of an organic dye, Rose Bengal (RB), which adhere to the particle surface, through the mechanism of luminescence energy transfer (LET).

Engineering a genetically-encoded SHG chromophore by electrostatic targeting to the membrane.

Although second harmonic generation (SHG) microscopy provides unique imaging advantages for voltage imaging and other biological applications, genetically-encoded SHG chromophores remain relatively unexplored. SHG only arises from non-centrosymmetric media, so an anisotropic arrangement of chromophores is essential to provide strong SHG signals. Here, inspired by the mechanism by which K-Ras4B associates with plasma membranes, we sought to achieve asymmetric arrangements of chromophores at the membrane-cytoplasm interface using the fluorescent protein mVenus. After adding a farnesylation motif to the C-terminus of mVenus, nine amino acids composing its β-barrel surface were replaced by lysine, forming an electrostatic patch. This protein (mVe9Knus-CVIM) was efficiently targeted to the plasma membrane in a geometrically defined manner and exhibited SHG in HEK293 cells. In agreement with its design, mVe9Knus-CVIM hyperpolarizability was oriented at a small angle (~7.3°) from the membrane normal. Genetically-encoded SHG chromophores could serve as a molecular platform for imaging membrane potential.

Detecting Subtle Plasma Membrane Perturbation in Living Cells Using Second Harmonic Generation Imaging

The requirement of center asymmetry for the creation of second harmonic generation (SHG) signals makes it an attractive technique for visualizing changes in interfacial layers such as the plasma membrane of biological cells. In this article, we explore the use of lipophilic SHG probes to detect minute perturbations in the plasma membrane. Three candidate probes, Di-4-ANEPPDHQ (Di-4), FM4-64, and all-trans-retinol, were evaluated for SHG effectiveness in Jurkat cells. Di-4 proved superior with both strong SHG signal and limited bleaching artifacts. To test whether rapid changes in membrane symmetry could be detected using SHG, we exposed cells to nanosecond-pulsed electric fields, which are believed to cause formation of nanopores in the plasma membrane. Upon nanosecond-pulsed electric fields exposure, we observed an instantaneous drop of ∼50% in SHG signal from the anodic pole of the cell. When compared to the simultaneously acquired fluorescence signals, it appears that the signal change was not due to the probe diffusing out of the membrane or changes in membrane potential or fluidity. We hypothesize that this loss in SHG signal is due to disruption in the interfacial nature of the membrane. The results show that SHG imaging has great potential as a tool for measuring rapid and subtle plasma membrane disturbance in living cells.

Non-Invasive Second Harmonic Generation (SHG) in Macroscopy (MacroSHG) as Bio-Diagnosis to Image Collagen Network Organization in Extracellular Matrix

In this work, we have considered a new multimodality imaging for macroscopy based on Second Harmonic Generation (SHG) method to monitor invasivelessly the matrix collagen. As the triple helicoidally structure of collagen molecules appearing as not centrosymetric, very organized and spatially oriented, collagen fibrils give rise to a very strong SHG signal and can be imaged without any exogenous dye. To integrate a multidimensional scale with a large field of view (non-sliced samples), we have adapted and validated an instrumental coupling between a two photon excitation laser and a macroscope to collect cartography of SHG signal. We introduced an index (F-SHG) based on decay time response measured by TCSPC for respectively Fluorescence (F) and Second Harmonic Generation (SHG) values. For various sample where protein collagen is the major component of extracellular matrix (vessel, skin, carotide vessel, rat femoral head cartilage, mouse tumor, human wharton’s jelly and rat tendon) or not (nacre), we compared the index distribution obtained with MacroSHG. In this work, we showed for the first time that multiscale large field imaging (Macroscopy) combined to Multimodality approaches (SHG-TCSPC) could be an innovative and non-invasive technique to detect and identify some biological interest molecules (collagen) in biomedical topics.