Superparamagnetic Composite Nanobeads Anchored with Molecular Glues for Ultrasensitive Label-free Proteomics

Mass spectrometry has emerged as a mainstream technique for label-free proteomics. However, proteomic coverage for trace samples is constrained by adsorption loss during repeated elution at sample pretreatment. Here, we demonstrated superparamagnetic composite nanoparticles functionalized with molecular glues (MGs) to enrich proteins in trace human biofluid. We showed high protein binding (>95 %) and recovery (≈90 %) rates by anchor-nanoparticles. We further proposed a Streamlined Workflow based on Anchor-nanoparticles for Proteomics (SWAP) method that enabled unbiased protein capture, protein digestion and pure peptides elution in one single tube. We demonstrated SWAP to quantify over 2500 protein groups with 100 HEK 293T cells. We adopted SWAP to profile proteomics with trace aqueous humor samples from cataract (n=15) and wet age-related macular degeneration (n=8) patients, and quantified ≈1400 proteins from 5 μL aqueous humor. SWAP simplifies sample preparation steps, minimizes adsorption loss and improves protein coverage for label-free proteomics with previous trace samples.     

Real-time Imaging of Nascent DNA in Live Cells by Monitoring the Fluorescence Lifetime of DNA-Incorporated Thiazole Orange-Modified Nucleotides

A modified 2′-deoxycytidine triphosphate derivative ( dC^TOTP ) bearing a thiazole orange moiety tethered via an oligoethylene glycol linker was designed and synthesized. The nucleotide was incorporated into DNA by DNA polymerases in vitro as well as in live cells. Upon incorporation of dC^TOTP into DNA, the thiazole orange moiety exhibited a fluorescence lifetime that differed significantly from the non-incorporated (i.e. free and non-covalently intercalated) forms of dC^TOTP . When dC^TOTP was delivered into live U-2 OS cells using a synthetic nucleoside triphosphate transporter, it allowed us to distinguish and monitor cells that were actively synthesizing DNA in real time, from the very first moments after the treatment. We anticipate that this probe could be used to study chromatin organization and dynamics.     

Mobility-Modulated Sequential Dissociation Analysis Enables Structural Lipidomics in Mass Spectrometry Imaging

Spatial lipidomics based on mass spectrometry imaging (MSI) is a powerful tool for fundamental biology studies and biomarker discovery. But the structure-resolving capability of MSI is limited because of the lack of multiplexed tandem mass spectrometry (MS/MS) method, primarily due to the small sample amount available from each pixel and the poor ion usage in MS/MS analysis. Here, we report a mobility-modulated sequential dissociation (MMSD) strategy for multiplex MS/MS imaging of distinct lipids from biological tissues. With ion mobility-enabled data-independent acquisition and automated spectrum deconvolution, MS/MS spectra of a large number of lipid species from each tissue pixel are acquired, at no expense of imaging speed. MMSD imaging is highlighted by MS/MS imaging of 24 structurally distinct lipids in the mouse brain and the revealing of the correlation of a structurally distinct phosphatidylethanolamine isomer (PE 18 : 1_18 : 1) from a human hepatocellular carcinoma (HCC) tissue. Mapping of structurally distinct lipid isomers is now enabled and spatial lipidomics becomes feasible for MSI.     

Expanding the Range of Bioorthogonal Tags for Multiplex Stimulated Raman Scattering Microscopy

Multiplex optical detection in live cells is challenging due to overlapping signals and poor signal-to-noise associated with some chemical reporters. To address this, the application of spectral phasor analysis to stimulated Raman scattering (SRS) microscopy for unmixing three bioorthogonal Raman probes within cells is reported. Triplex detection of a metallacarborane using the B−H stretch at 2480–2650 cm⁻¹, together with a bis-alkyne and deuterated fatty acid can be achieved within the cell-silent region of the Raman spectrum. When coupled to imaging in the high-wavenumber region of the cellular Raman spectrum, nine discrete regions of interest can be spectrally unmixed from the hyperspectral SRS dataset, demonstrating a new capability in the toolkit of multiplexed Raman imaging of live cells.     

Dimensional and Optoelectronic Tuning of Lead-free Perovskite Cs3Bi2I9−nBrn Single Crystals for Enhanced Hard X-ray Detection

Inorganic Bi-based perovskites have shown great potential in X-ray detection for their large absorption to X-rays, diverse low-dimensional structures, and eco-friendliness without toxic metals. However, they suffer from poor carrier transport properties compared to Pb-based perovskites. Here, we propose a mixed-halogen strategy to tune the structural dimensions and optoelectronic properties of Cs₃Bi₂I_9−nBr_n (0≤n≤9). Ten centimeter-sized single crystals are successfully grown by the Bridgman technique. Upon doping bromine to zero-dimensional Cs₃Bi₂I₉, the crystal transforms into a two-dimensional structure as the bromine content reaches Cs₃Bi₂I₈Br. Correspondingly, the optoelectronic properties are adjusted. Among these crystals, Cs₃Bi₂I₈Br exhibits negligible ion migration, moderate resistivity, and the best carrier transport capability. The sensitivities in 100 keV hard X-ray detection are 1.33×10⁴ and 1.74×10⁴ μC Gy_air⁻¹ cm⁻² at room temperature and 75 °C, respectively, which are the highest among all reported bismuth perovskites. Moreover, the lowest detection limit of 28.6 nGy_air s⁻¹ and ultralow dark current drift of 9.12×10⁻⁹ nA cm⁻¹ s⁻¹ V⁻¹ are obtained owing to the high ionic activation energy. Our work demonstrates that Br incorporation is an effective strategy to enhance the X-ray detection performance by tuning the dimensional and optoelectronic properties.     

SNR enhancement for composite application using multiple Doppler vibrometers based laser ultrasonic propagation imager

In recent years, the technology of using laser ultrasonic propagation imaging for damage visualization of composite structures were applied to real-world applications. Among many choices of sensor for the Ultrasonic Propagation Imager, the laser interferometry has several advantages: it is non-invasive, and portable, and with extraordinarily long-range measurement. However, the critical issue with interferometry sensing is its low signal-to-noise ratio (SNR), where the background noise can mask the damage-induced waves and making it impossible to identify the damages, especially in composite structures. In this paper, we propose a hardware-based SNR enhancement technique using multiple Laser Doppler Vibrometers (LDVs). The out-of-plane mode of ultrasonic signals are measured by multiple LDVs at a common sensing point and then averaged in real time. We showed that the SNR enhancement in experiments was consistent with the theoretical prediction, and also the test results showed a clear improvement for damage visualization of structures using Ultrasonic Wave Propagation Imaging and Ultrasonic Wavenumber Imaging algorithms.     

带有波前编码板的虹膜采集光学系统的设计

设计了一款采用波前编码技术的虹膜采集光学系统,以实现大景深、自然非约束状态下的成像.以传统的光学系统评价指标斯特利尔比和能量集中度作为波前编码系统的成像特征,利用ZEMAX软件对波前编码系统进行设计,权衡了离焦不敏感性和图像可恢复性.比较了带有立方相位编码板和带有修正项的立方相位编码板的虹膜采集系统以及传统系统的焦深延拓效果.发现采用立方相位编码板的虹膜采集光学系统的焦深延拓效果最佳,其焦深是传统系统的8倍.该系统增大了虹膜采集的距离,对于自然状态虹膜成像系统设计有一定的参考价值.     

基于单相机-双棱镜的小型三维指纹获取系统

提出了一种小型非接触式三维指纹获取系统.该系统采用单个CCD相机作为图像传感器,利用相机镜头结合双棱镜的成像装置在单帧CCD图像上获得立体图像对,再基于立体视觉原理重建三维指纹.整个系统封装后尺寸为12 mm×12 mm×10 mm.基于该系统进行了指纹获取实验,测量的均方根误差为0.022 mm.该系统操作简单且结构紧凑,为三维指纹的获取提供了一种小型化方案.     

Review: Recent applications of scanning electrochemical microscopy to the study of charge transfer kinetics

Scanning electrochemical microscopy (SECM) has been proven to be a valuable technique for the quantitative investigation and surface analysis of a wide range of processes that occur at interfaces. In particular, there is a great deal of interest in studying the kinetics of charge transfer characteristics at the solid/liquid and liquid/liquid interface. This overview outlines recent advances and applications of SECM to the investigation of charge transfer reactions at the solid/liquid interface and liquid/liquid interface.     

Partial least squares for discrimination in fMRI data

Multivariate methods for discrimination were used in the comparison of brain activation patterns between groups of cognitively normal women who are at either high or low Alzheimer's disease risk based on family history and apolipoprotein-E4 status. Linear discriminant analysis (LDA) was preceded by dimension reduction using principal component analysis (PCA), partial least squares (PLS) or a new oriented partial least squares (OrPLS) method. The aim was to identify a spatial pattern of functionally connected brain regions that was differentially expressed by the risk groups and yielded optimal classification accuracy. Multivariate dimension reduction is required prior to LDA when the data contain more feature variables than there are observations on individual subjects. Whereas PCA has been commonly used to identify covariance patterns in neuroimaging data, this approach only identifies gross variability and is not capable of distinguishing among-groups from within-groups variability. PLS and OrPLS provide a more focused dimension reduction by incorporating information on class structure and therefore lead to more parsimonious models for discrimination. Performance was evaluated in terms of the cross-validated misclassification rates. The results support the potential of using functional magnetic resonance imaging as an imaging biomarker or diagnostic tool to discriminate individuals with disease or high risk.