Assessing pathological features of breast cancer via the multimodal information of multiphoton and Raman imaging

For unveiling the pathological evolution of breast cancer, nonlinear multiphoton microscopic(MPM) and confocal Raman microspectral imaging(CRMI) techniques were both utilized to address the structural and constitutional characteristics of healthy(H), ductal carcinoma in situ(DCIS), and invasive ductal carcinoma(IDC) tissues. MPM-based techniques,including two-photon excited fluorescence(TPEF) and second harmonic generation(SHG), visualized label-free and the fine structure of breast tissue. Meanwhile, CRMI not only presented the chemical images of investigated samples with the K-mean cluster analysis method(KCA), but also pictured the distribution of components in the scanned area through univariate imaging. MPM images illustrated that the cancer cells first arranged around the basement membrane of the duct,then proliferated to fill the lumens of the duct, and finally broke through the basement membrane to infiltrate into the stroma.Although the Raman imaging failed to visualize the cell structure with high resolution, it explained spectroscopically the gradual increase of nucleic acid and protein components inside the ducts as cancer cells proliferated, and displayed the distribution pattern of each biological component during the evolution of breast cancer. Thus, the combination of MPM and CRMI provided new insights into the on-site pathological diagnosis of malignant breast cancer, also ensured technical support for the development of multimodal optical imaging techniques for precise histopathological analysis.

Particle velocity measurement of binary mixtures in the riser of a circulating fluidized bed by the combined use of electrostatic sensing and high-speed imaging

The flow dynamics of binary particle mixtures in the fluidized bed needs to be monitored in order to optimize the related industrial processes.In this paper,electrostatic sensing and high-speed imaging are applied to measure the velocities of polyethylene and sand particles in the binary particle mixtures in fluidization.Experimental studies were conducted on a lab-scale cold circulating fluidized bed.Correlation function between electrostatic signals from upstream and downstream electrodes placed along the riser shows two peaks that represent transit times for the two types of particles.To verify the above results,high-speed imaging was adopted to capture the flow images of particle mixtures.Particle Image Velocimetry and Particle Tracking Velocimetry algorithms were utilized to process the resulted images in order to measure the velocities of polyethylene and sand particles.The reasons for two-peak correlation functions are illustrated based on the frequency spectrums of the mono-solid-phase electrostatic signals and the velocity difference between polyethylene and sand particles.Finally,comparisons on the velocities obtained from electrostatic sensing and high-speed imaging demonstrate the electrostatic sensor can roughly estimate the particle velocity of binary particle mixtures in the near wall region of the circulating fluidized bed.

Compact and robust dual-color linearly polarized illumination source for three-photon fluorescence imaging

The miniaturized femtosecond laser in near infrared-Ⅱregion is the core equipment of threephoton microscopy.In this paper,we design a compact and robust illumination source that emits dual-color linearly polarized light for three-photon microscopy.Based on an all-polarizationmaintaining passive mode-locked fiber laser,we shift the center wavelength of the pulses to the 1.7m band utilizing cascade Raman effect,thereby generate dual-wavelength pulses.To enhance clarity,the two wavelengths are separated through the graded-index multimode fiber.Then we obtain the dual-pulse sequences with 1639.4 nm and 1683.7 nm wavelengths,920 fs pulse duration,and 23.75 MHz pulse repetition rate.The average power of the signal is 53.64mW,corresponding to a single pulse energy of 2.25 nJ.This illumination source can be further amplified and compressed for three-photon fluorescence imaging,especially dual-color three-photon fluorescence imaging,making it an ideal option for biomedical applications.

Wide-Field Vibrational Phase Contrast Imaging Based on Coherent Anti-Stokes Raman Scattering Holography

We propose and implement a wide-field vibrational phase contrast detection to obtain imaging of imaginary components of third-order nonlinear susceptibility in a coherent anti-Stokes Raman scattering （CARS） microscope with full suppression of the non-resonant background. This technique is based on the unique ability of recovering the phase of the generated CARS signal based on holographic recording. By capturing the phase distributions of the generated CARS field from the sample and from the environment under resonant illumination, we demonstrate the retrieval of imaginary components in the CARS microscope and achieve background free coherent Raman imaging.

In situ Raman imaging of high-temperature solid-state reactions in the CaSO4-SiO2 system

The deposition of mineral phases on the heat transfer surfaces of brown coal power plants may have a negative effect on power plant boilers. The paragenesis of these deposits contains information about the actual temperature prevailed during the combustion of lignite, if the temperature-dependences of distinct mineral transformations or reactions are known. Here, we report results of a sintering study (to ?1100℃) with samples containing anhydrite, quartz, and gehlenite, which are typical components of Rhenish lignite ashes. Thermal decompositions and solid-state reactions were analyzed (1) in situ and (2) both in situ and after quenching using confocal hyperspectral Raman imaging. This novel application of confocal Raman spectroscopy provides temperature-and time-resolved, 2-dimensional information about sintering processes with a micrometer-scale resolution. In the course of the sintering experiments with anhydrite and quartz with a weight ratio of 2:1 both polymorphs wollastonite and pseudowollastonite were identified in situ at about 920 and 1000℃, respectively. The formation of pseudowollastonite was thus observed about 120℃ below the phase transition temperature, demonstrating that it can form metastably. In addition,α′L-Ca2SiO4 was identified at about 1100℃. In samples containing equal weight fractions of anhydrite and quartz that were quenched after firing for 9h at about 1100℃,β-Ca2SiO4 (larnite) crystallized as rims around anhydrite grains and in direct contact to wollastonite. We furthermore observed that, depending on the ratio between quartz and anhydrite, wollastonite replaced quartz grains between 920 and 1100℃., i.e., the higher the quartz content, the lower the formation temperature of wollastonite.

Raman Micro-Imaging of the Coexistence of sⅠ and sⅡ Hydrates Formed from a Mixed Methane-Propane Gas in a Confined Space

Natural gas hydrate contains a specific amount of heavy hydrocarbons, such as ethane, propane, etc., aside from the primary guest gas of methane. Although the coexistence of two or even three hydrate structures has been discovered at several hydrate sites, the requisite formation mechanism is still not well understood. In-situ observation of the formation process of mixed methane-propane hydrate in a confined space was conducted using confocal Raman imaging microscopy. The Raman imaging results reveal that sI methane hydrate and sII mixed methane-propane hydrate are formed and coexist in the reaction system. In the confined space, the sI hydrate originates from the dissolved gas in water, while the sII hydrate is formed from free gas. The results obtained can help explain the coexistence of sI and sII hydrates found in natural hydrate samples, as well as providing insights into a possible dynamic scenario of hydrate reservoirs in geological history.

3D Confocal Raman Imaging of Oil-Rich Emulsion from Enzyme-Assisted Aqueous Extraction of Extruded Soybean Powder

The understanding of the structure morphology of oil-rich emulsion from enzyme-assisted extraction processing(EAEP)was a critical step to break the oil-rich emulsion structure in order to recover oil.Albeit EAEP method has been applied as an alternative way to conventional solvent extraction method,the structure morphology of oil-rich emulsion was still unclear.The current study aimed to investigate the structure morphology of oil-rich emulsion from EAEP using 3 D confocal Raman imaging technique.With increasing the enzymatic hydrolysis duration from 1 to 3 h,the stability of oil-rich emulsion was decreased as visualized in the 3 D confocal Raman images that the protein and oil were mixed together.The subsequent Raman spectrum analysis further revealed that the decreased stability of oil-rich emulsion was due to the protein aggregations via SS bonds or protein-lipid interactions.The conformational transfer in protein indicated the formation of a compact structure.

Stimulated Raman scattering tomography for rapid three-dimensional chemical imaging of cells and tissue

Three-dimensional(3D)imaging is essential for understanding intricate biological and biomedical systems,yet live cell and tissue imaging applications still face challenges due to constrained imaging speed and strong scattering in turbid media.Here,we present a unique phase-modulated stimulated Raman scattering tomography(PM-SRST)technique to achieve rapid label-free 3D chemical imaging in cells and tissue.To accomplish PM-SRST,we utilize a spatial light modulator to electronically manipulate the focused Stokes beam along the needle Bessel pump beam for SRS tomography without the need for mechanical z scanning.We demonstrate the rapid 3D imaging capability of PM-SRST by real-time monitoring of 3D Brownian motion of polystyrene beads in water with 8.5 Hz volume rate,as well as the instant biochemical responses to acetic acid stimulants in MCF-7 cells.Further,combining the Bessel pump beam with a longer wavelength Stokes beam(NIR-II window)provides a superior scattering resilient ability in PM-SRST,enabling rapid tomography in deeper tissue areas.The PM-SRST technique providestwofold enhancement in imaging depth in highly scattering media(e.g.,polymer beads phantom and biotissue like porcine skin and brain tissue)compared with conventional point-scan SRS.We also demonstrate the rapid 3D imaging ability of PM-SRST by observing the dynamic diffusion and uptake processes of deuterium oxide molecules into plant roots.The rapid PM-SRST developed can be used to facilitate label-free 3D chemical imaging of metabolic activities and functional dynamic processes of drug delivery and therapeutics in live cells and tissue.

Parallel Spectral-Domain Optical Coherence Tomography for Non-Scattering Object Imaging

The parallel spectral-domain optical coherence tomography（PSDOCT） is described for highspeed optical coherence tomography（OCT） without lateral scanning. In this setup, the self-elimination of auto-correlation（AC） interference algorithm was used for eradicating the AC interference and ghost images. However, when performed in free space OCT, this algorithm still generated a weak DC component. The algorithm was improved by adding the background intensity part to compensate for the mutual interference between object and reference arms. The results demonstrate that the DC component can be eradicated. Compared with conventional QCT and complex Fourier-domain optical coherence to- mography, the advantages of PSDOCT with the improved algorithm in free space are that it has no moving parts to generate consecutive phase shift, the structure of the object can be reconstructed immediately and automatically, and the speed is approximately 16 times faster than those of the other two in the same case.

The application of label-free imaging technologies in transdermal research for deeper mechanism revealing

The penetration behavior of topical substances in the skin not only relates to the transdermal delivery efficiency but also involves the safety and therapeutic effect of topical products,such as sunscreen and hair growth products.Researchers have tried to illustrate the transdermal process with diversified theories and technologies.Directly observing the distribution of topical substances on skin by characteristic imaging is the most convincing approach.Unfortunately,fluorescence labeling imaging,which is commonly used in biochemical research,is limited for transdermal research for most topical substances with a molecular mass less than 500 Da.Label-free imaging technologies possess the advantages of not requiring any macromolecular dyes,no tissue destruction and an extensive substance detection capability,which has enabled rapid development of such technologies in recent years and their introduction to biological tissue analysis,such as skin samples.Through the specific identification of topical substances and endogenous tissue components,label-free imaging technologies can provide abundant tissue distribution information,enrich theoretical and practical guidance for transdermal drug delivery systems.In this review,we expound the mechanisms and applications of the most popular label-free imaging technologies in transdermal research at present,compare their advantages and disadvantages,and forecast development prospects.

ACCURATE DETECTION OF HIGH-SPEED MULTI-TARGET VIDEO SEQUENCES MOTION REGIONS BASED ON RECONSTRUCTED BACKGROUND DIFFERENCE

The paper first discusses shortcomings of classical adjacent-frame difference. Sec ondly, based on the image energy and high order statistic(HOS) theory, background reconstruction constraints are setup. Under the help of block-processing technology, background is reconstructed quickly. Finally, background difference is used to detect motion regions instead of adjacent frame difference. The DSP based platform tests indicate the background can be recovered losslessly in about one second, and moving regions are not influenced by moving target speeds. The algorithm has important usage both in theory and applications.

Analysis of droplet transfer of pulsed MIG welding based on electrical signal and high-speed photography

In order to study how welding parameters affect welding quality and droplet transfer, a synchronous acquisition and analysis system is established to acquire and analyze electrical signal and instantaneous images of droplet transfer simultaneously, which is based on a self-developed soft-switching inverter. On the one hand, welding current and voltage signals are acquired and analyzed by a self-developed dynamic wavelet analyzer. On the other hand, images are filtered and optimized after they are captured by high-speed camera. The results show that instantaneous waveforms and statistical data of electrical signal contribute to make an overall assessment of welding quality, and that optimized high-speed images allow a visual and clear observation of droplet transfer process. The analysis of both waveforms and images leads to a further research on droplet transfer mechanism and provides a basis for precise control of droplet transfer.

Electromagnetic Tomography System for Defect Detection of High-Speed Rail Wheel

A novel electromagnetic tomography(EMT)system for defect detection of high-speed rail wheel is proposed,which differs from traditional electromagnetic tomography systems in its spatial arrangements of coils.A U-shaped sensor array was designed,and then a simulation model was built with the low frequency electromagnetic simulation software.Three different algorithms were applied to perform image reconstruction,therefore the defects can be detected from the reconstructed images.Based on the simulation results,an experimental system was built and image reconstruction were performed with the measured data.The reconstructed images obtained both from numerical simulation and experimental system indicated the locations of the defects of the wheel,which verified the feasibility of the EMT system and revealed its good application prospect in the future.

Recent advances in Raman spectroscopy for skin diagnosis

The skin is the largest organ in humans.It comprises about 16%of our body.Many diseases originate from the skin,including acne vulgaris,skin cancer,fungal skin disease,etc.As a common skin cancer in China,melanoma alone grows at year rate of nearly 4%.Therefore,it is crucial to develop an objective,reliable,accurate,non-invasive,and easy-to-use diagnostic method for skin diseases to support clinical decision-making.Raman spectroscopy is a highly specic imaging technique,which is sensitive,even to the single-cell level in skin diagnosis.Raman spectroscopy provides a pattern of signals with narrow bandwidths,making it a common and essential tool for researching individual characteristics of skin cells.Raman spectroscopy already has a number of clinical applications,including in thyroid,cervical and colorectal cancer.This review will introduce the advantages and recent developments in Raman spectroscopy,before focusing on the advances in skin diagnosis,including the advantages,methods,results,analysis,and notications.Finally,we discuss the current limitations and future progress of Raman spectroscopy in the context of skin diagnosis.

High-speed imaging observation on molten bridges of AgNi10 electrical contact material

The electrical contact-high-speed imaging experimental system was developed to investigate the molten bridge phenomena of AgNi10 electrical contact material.The dimension of molten bridges was measured along with the measurement of waveforms in the contact voltage under the load of direct current(DC) 6 V(8-20 A)and breaking speed of 50.0 mm·s^(-1).A part of the observed results was presented as well as surface morphology of the contacts after electrical contact behavior,which shows some interesting and new phenomena.Molten bridges and arc could exist simultaneously.The stable molten bridge looks like cylindrical shape and then becomes needle tip at its rupture,the diameter and length of molten bridges both increase with the increase in current and the growth gradient of the diameter is larger than that of the length.The morphology and elemental distribution of the contact surface are changed by the behavior of electrical contact.

Stimulated Raman scattering microscopy on biological cellular machinery

Beneting from the developments of advanced optical microscopy techniques,the mysteries of biological functions at the cellular and subcellular levels have been continuously revealed.Stimulated Raman scattering(SRS)microscopy is a rapidly growing technique that has attracted broad attentions and become a powerful tool for biology and biomedicine,largely thanks to its chemical specicity,high sensitivity and fast image speed.This review paper introduces the principles of SRS,discusses the technical developments and implementations of SRS microscopy,then highlights and summarizes its applications on biological cellular machinery andnally shares our visions of potential breakthroughs in the future.

共聚焦拉曼成像技术研究PE⁃LD/EVOH共混物的三维相结构

利用共聚焦拉曼成像技术研究了低密度聚乙烯(PE⁃LD)/乙烯⁃乙烯醇共聚物(EVOH)共混物中各相在压塑样品水平方向、深度方向和三维空间的分布情况,首次获得了PE⁃LD/EVOH共混物的三维相结构,并直观地将相区与共混物化学成分相对应。结果表明,PE⁃LD/EVOH共混物为不相容体系;当EVOH的质量分数为20%时,其作为分散相,以较规则的圆柱体形态分散于PE⁃LD基体中,圆柱直径尺寸在3~6μm范围内。加入马来酸酐接枝聚乙烯(PE⁃g⁃MAH)作为相容剂后,共混物的相态结构发生显著变化,分散相形状由规则变为不规则,截面的平均尺寸减小到约2μm,这说明PE⁃g⁃MAH可显著增强PE⁃LD与EVOH两组分间的界面相容性。

线扫式拉曼光谱仪的研制与生物医学应用进展

拉曼光谱技术是一种无损、高灵敏的分子成分检测技术,近年来在众多领域得到广泛应用,但由于逐点扫描的观测方式耗时较长,限制了在大面积样品拉曼扫描和成像中的应用。线扫式拉曼光谱通过线形激发光照明、同步采集线形样本区域拉曼信号的方式,相对于点扫式拉曼光谱仪成像速率获得了若干数量级的提升,从而使大面积、大数量样本具有时效性的拉曼光谱观测成为可能。本文对线扫式拉曼光谱仪的原理、搭建方式、系统结构和特点做了简要介绍,综述了其在细胞、组织观测等方面的应用进展,并对线扫式拉曼光谱性能改进及功能拓展技术进行了展望。

高温^(107)Ag^(5+)离子辐照后核石墨的截面微区拉曼表征与缺陷演化规律研究

在第四代反应堆中,核石墨作为慢化体和反射体材料服役于高温和高通量的快中子辐照环境中。快中子辐照会在核石墨中产生大量的弗伦克尔缺陷对。这些缺陷经过湮灭、扩散、最终形成更大的缺陷团簇,从而改变核石墨的微观结构,进而改变核石墨的宏观性能。因此,研究核石墨在高温辐照条件下的缺陷演化行为和机理对提高反应堆安全性具有重要意义。本研究采用30 MeV的^(107)Ag^(5+)离子在420℃下辐照IG-110核石墨来模拟核石墨在快中子辐照过程中的缺陷演化行为。通过微区拉曼光谱对IG-110核石墨截面结构进行表征,并对比不同深度处的拉曼光谱特征参数和辐照损伤剂量之间的关系,研究IG-110核石墨微观结构随辐照损伤剂量(Displacements Per Atom,DPA)的演化行为。研究结果表明,随着注量的增加,核石墨拉曼光谱的特征参数D峰高度与G峰高度比值(I_(D)/I_(G))、G峰半高宽(Full Width at Half Maximum of the G peak,FWHM(G))以及G峰的偏移量都显著增加。与^(58)Ni^(5+)辐照样品相比,相同辐照损伤剂量下,^(107)Ag^(5+)辐照的石墨拉曼光谱的I_(D)/I_(G)和FWHM(G)更大。相同的FWHM(G)下,^(107)Ag^(5+)辐照的石墨拉曼光谱的I_(D)/I_(G)比^(58)Ni^(5+)辐照样品大。这些结果说明更重的重离子辐照会在核石墨中引起更高速率的缺陷积累,从而更快地导致石墨晶粒尺寸变小,并促进纳米晶化进程。

基于拉曼光谱技术的聚合物材料制备与组成分布的研究进展

聚合物材料制备过程和聚合物中添加剂的含量及分布决定了材料的性能,拉曼光谱技术为相关研究提供了一种有效的表征手段。基于拉曼光谱的基本原理,介绍了表面增强拉曼光谱、拉曼光谱成像、激光显微共聚焦拉曼光谱等多种拉曼光谱衍生技术,总结了相关技术在聚合物加工和聚合反应过程中的应用,评述了聚合物中添加剂分布的检测,涵盖了乳化剂、染料以及其他添加剂的浓度、分布及迁移规律等,并对拉曼光谱衍生技术进行了展望。