Tutorial on methods for estimation of optical absorption and scattering properties of tissue.

Significance: The estimation of tissue optical properties using diffuse optics has found a range of applications in disease detection, therapy monitoring, and general health care. Biomarkers derived from the estimated optical absorption and scattering coefficients can reflect the underlying progression of many biological processes in tissues. Aim: Complex light-tissue interactions make it challenging to disentangle the absorption and scattering coefficients, so dedicated measurement systems are required. We aim to help readers understand the measurement principles and practical considerations needed when choosing between different estimation methods based on diffuse optics. Approach: The estimation methods can be categorized as: steady state, time domain, time frequency domain (FD), spatial domain, and spatial FD. The experimental measurements are coupled with models of light-tissue interactions, which enable inverse solutions for the absorption and scattering coefficients from the measured tissue reflectance and/or transmittance. Results: The estimation of tissue optical properties has been applied to characterize a variety of ex vivo and in vivo tissues, as well as tissue-mimicking phantoms. Choosing a specific estimation method for a certain application has to trade-off its advantages and limitations. Conclusion: Optical absorption and scattering property estimation is an increasingly important and accessible approach for medical diagnosis and health monitoring.

Polynomial chaos expansion of SAR and temperature increase variability in 3 T MRI due to stochastic input data.

Objective.Numerical simulations are largely adopted to estimate dosimetric quantities, e.g. specific absorption rate (SAR) and temperature increase, in tissues to assess the patient exposure to the radiofrequency (RF) field generated during magnetic resonance imaging (MRI). Simulations rely on reference anatomical human models and tabulated data of electromagnetic and thermal properties of biological tissues. However, concerns may arise about the applicability of the computed results to any phenotype, introducing a significant degree of freedom in the simulation input data. In addition, simulation input data can be affected by uncertainty in relative positioning of the anatomical model with respect to the RF coil. The objective of this work is the to estimate the variability of SAR and temperature increase at 3 T head MRI due to different sources of variability in input data, with the final aim to associate a global uncertainty to the dosimetric outcomes.Approach.A stochastic approach based on arbitrary Polynomial Chaos Expansion is used to evaluate the effects of several input variability's (anatomy, tissue properties, body position) on dosimetric outputs, referring to head imaging with a 3 T MRI scanner.Main results.It is found that head anatomy is the prevailing source of variability for the considered dosimetric quantities, rather than the variability due to tissue properties and head positioning. From knowledge of the variability of the dosimetric quantities, an uncertainty can be attributed to the results obtained using a generic anatomical head model when SAR and temperature increase values are compared with safety exposure limits.Significance.This work associates a global uncertainty to SAR and temperature increase predictions, to be considered when comparing the numerically evaluated dosimetric quantities with reference exposure limits. The adopted methodology can be extended to other exposure scenarios for MRI safety purposes.

Absorption of 5G sub-6 GHz electromagnetic radiation from base station to male reproduction system.

BACKGROUND: The impact of electromagnetic radiation from communication on the male reproductive system has emerged as a significant concern in public health. A notable distinction of the 5G sub-6 GHz band, compared to traditional 2G, 3G, and 4G frequency bands, is the inclusion of higher frequency bands. This has raised public concerns regarding the potential effects of these higher frequencies on organisms, particularly their reproductive systems. While it is imperative to investigate the biological effects and potential risks associated with these new frequency bands in laboratory settings, comparing and evaluating differences between various frequency bands remain challenging due to the absence of standardized parameters such as exposure conditions and duration. In contrast, dose assessment offers a simpler and more reliable approach. MATERIALS AND METHODS: The dose assessment method was employed in this study to investigate the risks associated with sub-6 GHz electromagnetic radiation from 5G base stations on the male reproductive system. A classical human body model (Duke) was utilized, and an electromagnetic simulation environment was established based on the actual polarization direction of the exposed base stations and various body postures. This research explored the effects of field direction, posture, public population, and frequency on the specific absorption rate of the reproductive system. RESULTS AND CONCLUSIONS: While maintaining the same level of exposure, a higher frequency results in a reduced dosage on reproductive system. Further analysis reveals that, considering the public exposure threshold, the employment of higher frequency bands in 5G sub-6 GHz does not present a greater dosage on reproductive system compared to lower frequency bands. Consequently, with regard to dosage, there is no need for excessive concern among the general public regarding the impact of electromagnetic radiation emitted by 5G base stations operating below 6 GHz on male reproductive health.

Electromagnetic interference shielding of flexible carboxymethyl cellulose/MWCNT@Fe3O4 composite film with ultralow reflection loss.

Nowadays, various high-performance electromagnetic interference (EMI) shielding materials have enormous application potential in electronic field. However, traditional EMI shielding materials often have high conductivity, resulting in the serious mismatch between the impedance of the material surface and the free space, which will cause a large amount of reflection of electromagnetic (EM) waves, leading to secondary reflection pollution. In this paper, we report a novel flexible EMI shielding composite film with extremely low reflection loss and efficient EM wave absorption, which was prepared by assisted self-assembly based on simple vacuum filtration using carboxymethyl cellulose as the matrix and MWCNT@Fe3O4 synthesized by chemical coprecipitation as the composite functional filler. By adjusting the Fe3O4 coating degree of MWCNTs in the filler, the composite film achieved the construction of a conductive network with high Fe3O4 content. Benefit by the good adaptability of conductivity and magnetic permeability, the composite film has good impedance matching ability and microwave absorption performance. The reflection loss of the composite film with the thickness of 28 µm in the X-band was only 0.23 dB, accounting for 1.7 % of the total loss. This work provides new insights for the development of EMI materials and effective mitigation secondary EM wave reflection pollution.

Immunogenic Radiation Therapy for Enhanced Antitumor Immunity via a Core-Shell Nanosensitizer-Mediated Immunosuppressive Tumor Microenvironment Modulation.

Due to the immunosuppressive tumor microenvironment (TME) and weak radiation absorption, the immune response triggered by radiation therapy (RT) is limited. Herein, a core-shell nanosensitizer UiO@MnS (denoted as UM) was genuinely constructed for the amplification of RT efficacy and induction of immunogenicity via integrating MnS-reprogrammed TME with Hf-based UiO-sensitized RT. The acid-sensitive MnS would produce H2S under acidic TME to improve oxygenation through inhibition mitochondrial respiration and reducing metabolic oxygen consumption, leading to decreased HIF-1α expression and enhanced radiosensitization. In addition, the generated H2S inhibited the catalase activity to increase the H2O2 level, which subsequently enhanced the Mn2+-mediated Fenton-like reaction, resulting in G2/M cell cycle arrest to improve the cellular sensitivity for radiation. This impressive tumor oxygenation, cell cycle arrest, and radiosensitization procedure boosted RT efficacy and resulted in strong antitumor immunogenicity. Taken together, combining the immunosuppressive TME modulation with a sensitizing radiation strategy shows great promise for magnifying immunogenic RT outputs.

Core-Shell Structured Silica-Coated Iron Nanowires Composites for Enhanced Electromagnetic Wave Absorption Properties.

In this study, we successfully prepared core-shell heterostructured nanocomposites (Fe NWs@SiO2), with ferromagnetic nanowires (Fe NWs) as the core and silica (SiO2) as the shell. The composites exhibited enhanced electromagnetic wave absorption and oxidation resistance and were synthesized using a simple liquid-phase hydrolysis reaction. We tested and analyzed the microwave absorption properties of Fe NWs@SiO2 composites with varied filling rates (mass fractions of 10 wt%, 30 wt%, and 50 wt% after mixing with paraffin). The results showed that the sample filled with 50 wt% had the best comprehensive performance. At the matching thickness of 7.25 mm, the minimum reflection loss (RLmin) could reach -54.88 dB at 13.52 GHz and the effective absorption bandwidth (EAB, RL < -10 dB) could reach 2.88 GHz in the range of 8.96-17.12 GHz. Enhanced microwave absorption performance of the core-shell structured Fe NWs@SiO2 composites could be attributed to the magnetic loss of the composite, the core-shell heterogeneous interface polarization effect, and the small-scale effect induced by the one-dimensional structure. Theoretically, this research provided Fe NWs@SiO2 composites with highly absorbent and antioxidant core-shell structures for future practical applications.

Advances and mechanisms in polymer composites toward thermal conduction and electromagnetic wave absorption.

Polymer composites have essential applications in electronics due to their versatility, stable performance, and processability. However, with the increasing miniaturization and high power of electronics in the 5G era, there are significant challenges related to heat accumulation and electromagnetic wave (EMW) radiation in narrow spaces. Traditional solutions involve using either thermally conductive or EMW absorbing polymer composites, but these fail to meet the demand for multi-functional integrated materials in electronics. Therefore, designing thermal conduction and EMW absorption integrated polymer composites has become essential to solve the problems of heat accumulation and electromagnetic pollution in electronics and adapt to its development trend. Researchers have developed different approaches to fabricate thermal conduction and EMW absorption integrated polymer composites, including integrating functional fillers with both thermal conduction and EMW absorption functions and innovating processing methods. This review summarizes the latest research progress, factors that affect performance, and the mechanisms of thermal conduction and EMW absorption integrated polymer composites. The review also discusses problems that limit the development of these composites and potential solutions and development directions. The aim of this review is to provide references for the development of thermal conduction and EMW absorption integrated polymer composites.

Development of a charged model of the SARS-CoV-2 viral surface.

A recent study provided experimental evidence of inactivation of viral activity after radio-frequency (RF) exposures in the 6-12 GHz band that was hypothesized to be caused by vibrations of an acoustic dipole mode in the virus that excited the viral membrane to failure. Here, we develop an atomic-scale molecular dynamics (MD) model of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) viral surface to estimate the electric fields necessary to rupture the viral membrane via dipole shaking of the virus. We computed the absorption spectrum of the system via unbiased MD simulations and found no particular strong absorption in the GHz band. We investigated the mechanical resiliency of the viral membrane by introducing uniaxial strains in the system and observed no pore formation in the membrane for strains up to 50%. Because the computed absorption spectrum was found to be essentially flat, and the strain required to break the viral membrane was >0.5, the field strength associated with rupture of the virus was greater than the dielectric breakdown value of air. Thus, RF disinfection of enveloped viruses would occur only once sufficient heat was transferred to the virus via a thermal mechanism and not by direct action (shaking) of the RF field oscillations on the viral membrane.

Palladium-modified TiO2 films in a photocatalytic microreactor: evaluation of radiation absorption properties and pollutant degradation efficiency.

Pure (TiO2) and 0.1 nominal atomic percent of palladium-modified TiO2 (Pd-TiO2) films were synthesized via a sol-gel method and compared through their physicochemical properties and photocatalytic activity in the degradation of an emerging contaminant, 17-α-ethinylestradiol (EE2). The activity of the films was studied using a continuous flow, planar microreactor under simulated sunlight. Catalysts characterization included X-ray diffraction, UV-Visible diffuse reflectance and transmittance spectroscopy, atomic force microscopy, transmission electron microscopy, Raman spectroscopy, N2 physisorption analysis, and X-ray photoelectron spectroscopy. The modification of TiO2 with palladium confined the size of anatase phase crystallites, increased the specific surface area and improved radiation absorption. PdO domains on TiO2 were observed. In all the tested conditions, higher conversion of EE2 was achieved with the Pd-TiO2 film compared with the TiO2 film, presenting an 80% increase in the reaction rate. The performance of the catalytic films was also assessed by the calculation of two efficiency parameters: radiation absorption efficiency and quantum efficiency of reaction. The Pd-TiO2 film showed a notable enhancement of the absorption of the incident radiation and a more efficient utilization of the absorbed photons to degrade the target pollutant.

Construction of Dual-Shell Mo2C/C Microsphere towards Efficient Electromagnetic Wave Absorption.

Carbon-based carbides have attracted tremendous attention for electromagnetic energy attenuation due to their adjustable dielectric properties, oxidation resistance, and good chemical stability. Herein, we reasonably regulate the growth of dopamine hydrochloride on the surface of the Mo-glycerate (Mo-GL) microsphere and then transform the resultant Mo-polydopamine (Mo-PD) microsphere into a dual-shell Mo2C/C (DS-Mo2C/C) microsphere in a high-temperature pyrolysis process under an inert atmosphere. It is found that the pyrolysis temperature plays an important role in the graphitization degree of the carbon matrix and internal architecture. The fabrication of a dual-shell structure can be propitious to the optimization of impedance matching, and the introduction of Mo2C nanoparticles also prompts the accumulation of polarization loss. When the pyrolysis temperature reaches 800 °C, the optimized composite of DS-Mo2C/C-800 exhibits good EM absorption performance in the frequency range of 2.0-18.0 GHz. DS-Mo2C/C-800's qualified bandwidth can reach 4.4 GHz at a matching thickness of 1.5 mm, and the integrated qualified bandwidth (QBW) even exceeds 14.5 GHz with a thickness range of 1.5-5.0 mm. The positive effects of the dual-shell structure and Mo2C nanoparticles on EM energy attenuation may render the DS-Mo2C/C microsphere as a promising candidate for lightweight and broad bandwidth EM absorption materials in the future.

A novel and effective technique to reduce electromagnetic radiation absorption on biotic components at 2.45 GHz.

A strong evidence of the effects of radiation absorption on the living community together with a better solution to reduce the radiation intensity without compromising the usage of wireless communication systems is presented. This study analyses the radiation effects on living things and validates the proposed techniques for specific absorption rate (SAR) value reduction at 2.45 GHz. To reduce these radiation impacts on the living community, proper shielding from the radiation and effectively reorienting antenna radiation patterns are the solutions suggested. An analogous antenna configuration in wireless communication systems - a coplanar waveguide fed loop antenna is considered and an open loop resonator (OLR) optimized in ANSYS HFSS at 2.45 GHz is incorporated on the back side of the proposed antenna for achieving SAR value reduction. Theoretical and experimental validation is carried out by measuring the variation in absorption power on each vegetable sample using vector network analyzer E5080A. The existence of OLR on the back side of the antenna reduces the absorption power upto 2 dB. From experimental validation, the proposed technique provides 88% to 98% reduction in SAR value when tested in each sample. Along with this OLR exhibits the capability to enhance the shielding characteristics to the controlled environment of experimental setup for analyzing the stages of seed germination, which helps in reducing the reported radiation effects and growth retardation. The proposed method of EMR reduction with miniaturized planar resonator can be effectively used in the communication systems operating at 2.45 GHz for creating a reduced radiation environment.

Core-Shell Structured SiO2@NiFe LDH Composite for Broadband Electromagnetic Wave Absorption.

In this work, a novel core-shell structure material, NiFe layered double hydroxide (NiFe LDH) loaded on SiO2 microspheres (SiO2@NiFe LDH), was synthesized by a one-step hydrothermal method, and the spontaneous electrostatic self-assembly process. The morphology, structure, and microwave absorption properties of SiO2@NiFe LDH nanocomposites with different NiFe element ratios were systematically investigated. The results show that the sample of SiO2@NiFe LDH-3 nanocomposite has excellent microwave absorption properties. It exhibits broadband effective absorption bandwidth (RL < −10 dB) of 8.24 GHz (from 9.76 GHz to 18.0 GHz) and the reflection loss is −53.78 dB at the matched thickness of 6.95 mm. It is expected that this SiO2@NiFe-LDH core-shell structural material can be used as a promising non-precious, metal-based material microwave absorber to eliminate electromagnetic wave contamination.

Corneal riboflavin gradients and UV-absorption characteristics after topical application of riboflavin in concentrations ranging from 0.1 to 0.5.

Avoiding damage of the endothelial cells, especially in thin corneas, remains a challenge in corneal collagen crosslinking (CXL). Knowledge of the riboflavin gradients and the UV absorption characteristics after topical application of riboflavin in concentrations ranging from 0.1% to 0.5% could optimize the treatment. In this study, we present a model to calculate the UV-intensity depending on the corneal thickness. Ten groups of de-epithelialized porcine corneas were divided into 2 subgroups. Five groups received an imbibition of 10 min and the other five groups for 30 min. The applied riboflavin concentrations were 0.1%, 0.2%, 0.3%, 0.4% and 0.5% diluted in a 15% dextran solution for each subgroup. After the imbibition process, two-photon fluorescence microscopy was used to determine fluorescence intensity, which was compared to samples after saturation, yielding the absolute riboflavin concentration gradient of the cornea. The extinction coefficient of riboflavin solutions was measured using a spectrophotometer. Combining the obtained riboflavin concentrations and the extinction coefficients, a depth-dependent UV-intensity profile was calculated for each group. With increasing corneal depth, the riboflavin concentration decreased for all imbibition solutions and application times. The diffusion coefficients of 10 min imbibition time were higher than for 30 min. A higher RF concentration and a longer imbibition time resulted in higher UV-absorption and a lower UV-intensity in the depth of the cornea. Calculated UV-transmission was 6 percentage points lower compared to the measured transmission. By increasing the riboflavin concentration of the imbibition solution, a substantially higher UV-absorption inside the cornea is achieved. This offers a simple treatment option to control the depth of crosslinking e.g. in thin corneas, resulting in a lower risk of endothelial damage.

Physicochemical, antioxidant properties of carotenoids and its optoelectronic and interaction studies with chlorophyll pigments.

The physicochemical and antioxidant properties of seven carotenoids: antheraxanthin, ß-carotene, neoxanthin, peridinin, violaxanthin, xanthrophyll and zeaxanthin were studied by theoretical means. Then the Optoelectronic properties and interaction of chlorophyll-carotenoid complexes are analysed by TDDFT and IGMPLOT. Global reactivity descriptors for carotenoids and chlorophyll (Chla, Chlb) are calculated via conceptual density functional theory (CDFT). The higher HOMO-LUMO (HL) gap indicated structural stability of carotenoid, chlorophyll and chlorophyll-carotenoid complexes. The chemical hardness for carotenoids and Chlorophyll is found to be lower in the solvent medium than in the gas phase. Results showed that carotenoids can be used as good reactive nucleophile due to lower µ and ω. As proton affinities (PAs) are much lower than the bond dissociation enthalpies (BDEs), it is anticipated that direct antioxidant activity in these carotenoids is mainly due to the sequential proton loss electron transfer (SPLET) mechanism with dominant solvent effects. Also lower PAs of carotenoid suggest that antioxidant activity by the SPLET mechanism should be a result of a balance between proclivities to transfer protons. Reaction rate constant with Transition-State Theory (TST) were estimated for carotenoid-Chlorophyll complexes in gas phase. Time dependent Density Functional Theory (TDDFT) showed that all the chlorophyll (Chla, Chlb)-carotenoid complexes show absorption wavelength in the visible region. The lower S1-T1 adiabatic energy gap indicated ISC transition from S1 to T1 state.

3D printing fluorescent material with tunable optical properties.

The 3D printing of fluorescent materials could help develop, validate, and translate imaging technologies, including systems for fluorescence-guided surgery. Despite advances in 3D printing techniques for optical targets, no comprehensive method has been demonstrated for the simultaneous incorporation of fluorophores and fine-tuning of absorption and scattering properties. Here, we introduce a photopolymer-based 3D printing method for manufacturing fluorescent material with tunable optical properties. The results demonstrate the ability to 3D print various individual fluorophores at reasonably high fluorescence yields, including IR-125, quantum dots, methylene blue, and rhodamine 590. Furthermore, tuning of the absorption and reduced scattering coefficients is demonstrated within the relevant mamalian soft tissue coefficient ranges of 0.005-0.05 mm-1 and 0.2-1.5 mm-1, respectively. Fabrication of fluorophore-doped biomimicking and complex geometric structures validated the ability to print feature sizes less than 200 µm. The presented methods and optical characterization techniques provide the foundation for the manufacturing of solid 3D printed fluorescent structures, with direct relevance to biomedical optics and the broad adoption of fast manufacturing methods in fluorescence imaging.

A Halomanganates(II) with P,P'-Diprotonated Bis(2-Diphenylphosphinophenyl)ether: Wavelength-Excitation Dependence of the Quantum Yield and Role of the Non-Covalent Interactions.

A [H2DPEphos][MnX4] [X = Br, Cl] tetrahalomanganates(II) with P,P'-diprotonated bis[2-(diphenylphosphino)phenyl]ether cation has been designed and investigated in photophysics and EPR terms. The complexes exhibit a green luminescence resulted from the Mn(II) d-d transitions (4T1→6A1) with the wavelength-excitation dependence of the quantum yield. The solid [H2DPEphos][MnBr4] complex exhibits a bright green phosphorescence (λmax = 515 nm) with the high luminescence quantum yield depending on the excitation energy whereas the solid [H2DPEphos][MnCl4] complex exhibits a very weak phosphorescence (λmax = 523 nm). The unexpected shorter luminescence lifetime for the [H2DPEphos][MnCl4] than for the [H2DPEphos][MnBr4] at 300 K can be a result of the higher non-radiative relaxation contribution. On the one hand, the non-covalent PH…X(Mn) interactions quench the manganese(II) luminescence. On the other hand, the PH…X(Mn) interactions are a pathway of the excitation transfer from [H2DPEphos]2+ to [MnX4]2-.

Dosimetry during adjuvant 131I therapy in patients with differentiated thyroid cancer-clinical implications.

The activity of radioiodine (131I) used in adjuvant therapy for thyroid cancer ranges between 30 mCi (1.1 GBq) and 150 mCi (5.5 GBq). Dosimetry based on Marinelli's formula, taking into consideration the absorbed dose in the postoperative tumour bed (D) should systematise the determination of 131I activity. Retrospective analysis of 57 patients with differentiated thyroid cancer (DTC) after thyreidectomy and adjuvant 131I therapy with the fixed activity of 3.7 GBq. In order to calculate D from Marinelli's formula, the authors took into account, among other things, repeated dosimetry measurements (after 6, 24, and 72 h) made during scintigraphy and after administration of the therapeutic activity or radioiodine. In 75% of the patients, the values of D were > 300 Gy (i.e. above the value recommended by current guidelines). In just 16% of the patients, the obtained values fell between 250 and 300 Gy, whereas in 9% of the patients, the value of D was < 250 Gy. The therapy was successful for all the patients (stimulated Tg < 1 ng/ml and 131I uptake < 0.1% in the thyroid bed in follow-up examination). Dosimetry during adjuvant 131I therapy makes it possible to diversify the therapeutic activities of 131I in order to obtain a uniform value of D.

Determination of the characteristic inactivation fluence for SARS-CoV-2 under UV-C radiation considering light absorption in culture media.

The optical absorption coefficient of culture media is critical for the survival analysis of pathogens under optical irradiation. The quality of the results obtained from experiments relies on the optical analysis of the spatial distribution of fluence which also depends on the geometry of the sample. In this contribution, we consider both the geometrical shape and the culture medium's absorption coefficient to evaluate how the spatial distribution of optical radiation affects pathogens/viruses. In this work, we exposed SARS-CoV-2 to UV-C radiation ([Formula: see text] = 254 nm) and we calculated-considering the influence of the optical absorption of the culture medium-a characteristic inactivation fluence of [Formula: see text] = 4.7 J/m2, or an equivalent 10% survival (D90 dose) of 10.8 J/m2. Experimentally, we diluted the virus into sessile drops of Dulbecco's Modified Eagle Medium to evaluate pathogen activity after controlled doses of UV irradiation. To validate the optical absorption mode, we carried out an additional experiment where we varied droplet size. Our model-including optical absorption and geometrical considerations-provides robust results among a variety of experimental situations, and represents our experimental conditions more accurately. These results will help to evaluate the capability of UV disinfecting strategies applied to a variety of everyday situations, including the case of micro-droplets generated by respiratory functions.

SAR and temperature distributions in a database of realistic human models for 7 T cardiac imaging.

PURPOSE: To investigate inter-subject variability of B1+ , SAR and temperature rise in a database of human models using a local transmit array for 7 T cardiac imaging. METHODS: Dixon images were acquired of 14 subjects and segmented in dielectric models with an eight-channel local transmit array positioned around the torso for cardiac imaging. EM simulations were done to calculate SAR distributions. Based on the SAR distributions, temperature simulations were performed for exposure times of 6 min and 30 min. Peak local SAR and temperature rise levels were calculated for different RF shim settings. A statistical analysis of the resulting peak local SAR and temperature rise levels was performed to arrive at safe power limits. RESULTS: For RF shim vectors with random phase and uniformly distributed power, a safe average power limit of 35.7 W was determined (first level controlled mode). When RF amplitude and phase shimming was performed on the heart, a safe average power limit of 35.0 W was found. According to Pennes' model, our numerical study suggests a very low probability of exceeding the absolute local temperature limit of 40 °C for a total exposure time of 6 min and a peak local SAR of 20 W/kg. For a 30 min exposure time at 20 W/kg, it was shown that the absolute temperature limit can be exceeded in the case where perfusion does not change with temperature. CONCLUSION: Safe power constraints were found for 7 T cardiac imaging with an eight-channel local transmit array, while considering the inter-subject variability of B1+ , SAR and temperature rise.

Label-Free Multiphoton Microscopy: Much More Than Fancy Images.

Multiphoton microscopy has recently passed the milestone of its first 30 years of activity in biomedical research. The growing interest around this approach has led to a variety of applications from basic research to clinical practice. Moreover, this technique offers the advantage of label-free multiphoton imaging to analyze samples without staining processes and the need for a dedicated system. Here, we review the state of the art of label-free techniques; then, we focus on two-photon autofluorescence as well as second and third harmonic generation, describing physical and technical characteristics. We summarize some successful applications to a plethora of biomedical research fields and samples, underlying the versatility of this technique. A paragraph is dedicated to an overview of sample preparation, which is a crucial step in every microscopy experiment. Afterwards, we provide a detailed review analysis of the main quantitative methods to extract important information and parameters from acquired images using second harmonic generation. Lastly, we discuss advantages, limitations, and future perspectives in label-free multiphoton microscopy.