Terahertz s-SNOM Imaging of a Single Cell with Nanoscale Resolution.

Terahertz scattering scanning near-field optical microscopy is a robust spectral detection technique with a nanoscale resolution. However, there are still major challenges in investigating the heterogeneity of cell membrane components in individual cells. Here, we present a novel and comprehensive analytical approach for detecting and investigating heterogeneity in cell membrane components at the single-cell level. In comparison to the resolution of the topographical atomic force microscopy image, the spatial resolution of the terahertz near-field amplitude image is 3 times that of the former. This ultrafine resolution enables the compositional distribution in the cell membrane, such as the distribution of extracellular vesicles, to be finely characterized. Furthermore, via extraction of the near-field absorption images at specific frequencies, the visualization and compositional difference analysis of cell membrane components can be presented in detail. These findings have significant implications for the intuitive and visual analysis of cell development and disease evolutionary pathways.

Consumer electronics based smart technologies for enhanced terahertz healthcare having an integration of split learning with medical imaging.

The proposed work contains three major contribution, such as smart data collection, optimized training algorithm and integrating Bayesian approach with split learning to make privacy of the patent data. By integrating consumer electronics device such as wearable devices, and the Internet of Things (IoT) taking THz image, perform EM algorithm as training, used newly proposed slit learning method the technology promises enhanced imaging depth and improved tissue contrast, thereby enabling early and accurate disease detection the breast cancer disease. In our hybrid algorithm, the breast cancer model achieves an accuracy of 97.5 percent over 100 epochs, surpassing the less accurate old models which required a higher number of epochs, such as 165.

Near-Field Terahertz Morphological Reconstruction Nanoscopy for Subsurface Imaging of Protein Layers.

Protein layers formed on solid surfaces have important applications in various fields. High-resolution characterization of the morphological structures of protein forms in the process of developing protein layers has significant implications for the control of the layer's quality as well as for the evaluation of the layer's performance. However, it remains challenging to precisely characterize all possible morphological structures of protein in various forms, including individuals, networks, and layers involved in the formation of protein layers with currently available methods. Here, we report a terahertz (THz) morphological reconstruction nanoscopy (THz-MRN), which can reveal the nanoscale three-dimensional structural information on a protein sample from its THz near-field image by exploiting an extended finite dipole model for a thin sample. THz-MRN allows for both surface imaging and subsurface imaging with a vertical resolution of ∼0.5 nm, enabling the characterization of various forms of proteins at the single-molecule level. We demonstrate the imaging and morphological reconstruction of single immunoglobulin G (IgG) molecules, their networks, a monolayer, and a heterogeneous double layer comprising an IgG monolayer and a horseradish peroxidase-conjugated anti-IgG layer. The established THz-MRN presents a useful approach for the label-free and nondestructive study of the formation of protein layers.

Enhanced in-situ liquid transport investigation setup for pharmaceutical tablet disintegration analysis using terahertz radiation.

The disintegration process of pharmaceutical solid dosage forms commences on contact with the dissolution medium and continues with subsequent spontaneous imbibition of the medium in the tablet matrix. Identifying the location of the liquid front in situ during imbibition, therefore, plays a significant role in understanding and modelling the disintegration process. Terahertz pulsed imaging (TPI) technology can be used to investigate this process by its ability to penetrate and identify the liquid front in pharmaceutical tablets. However, previous studies were limited to samples suitable for a flow cell environment, i.e. flat cylindrical disk shapes; thus, most commercial tablets could only be measured with prior destructive sample preparation. This study presents a new experimental setup named open immersion to measure a wide range of pharmaceutical tablets in their intact form. Besides, a series of data processing techniques to extract subtle features of the advancing liquid front are designed and utilised, effectively increasing the maximum thickness of tablets that can be analysed. We used the new method and successfully measured the liquid ingress profiles for a set of oval convex tablets prepared from a complex eroding immediate-release formulation.

Studying the dissolution of immediate release film coating using terahertz pulsed imaging.

Coated tablets introduce complexity to the dissolution process, even with readily soluble immediate release coating layers. Therefore, a more detailed understanding of the physical steps involved in the dissolution process can improve the efficiency of formulation and process design. The current study uses terahertz pulsed imaging to visualise the hydration process of microcrystalline cellulose (MCC) tablet cores that were film coated with an immediate release coating formulation upon exposure to the dissolution medium. Film coated tablets that were prepared from three levels of core porosity (10%, 20% and 30%) and with coating thickness in the range of 30µm to 250µm were investigated. It was possible to resolve and quantify the distinct stages of wetting of the coating layer, swelling of the MCC particles at the core surface, and dissolution of the coating layer followed by the ingress of dissolution media into the tablet core. The liquid transport process through the coating layer was highly consistent and scalable. The penetration rate through the coating layer and the tablet core both strongly depended on coating thickness and core porosity.

Research progress on the application of spectral imaging technology in pharmaceutical tablet analysis.

Tablet as a traditional dosage form in pharmacy has the advantages of accurate dosage, ideal dissolution and bioavailability, convenient to carry and transport. The most concerned tablet quality attributes include active pharmaceutical ingredient (API) contents and polymorphic forms, components distribution, hardness, density, coating state, dissolution behavior, etc., which greatly affect the bioavailability and consistency of tablet final products. In the pharmaceutical industry, there are usually industry standard methods to analyze the tablet quality attributes. However, these methods are generally time-consuming and laborious, and lack a comprehensive understanding of the properties of tablets, such as spatial information. In recent years, spectral imaging technology makes up for the shortcomings of traditional tablet analysis methods because it provides non-contact and rich information in time and space. As a promising technology to replace the traditional tablet analysis methods, it has attracted more and more attention. The present paper briefly describes a series of spectral imaging techniques and their applications in tablet analysis. Finally, the possible application prospect of this technology and the deficiencies that need to be improved were also prospected.

Towards super-resolved terahertz microscopy for cellular imaging.

Biomedical imaging includes the use of a variety of techniques to study organs and tissues. Some of the possible imaging modalities are more spread at clinical level (CT, MRI, PET), while others, such as light and electron microscopy are preferred in life sciences research. The choice of the imaging modalities can be based on the capability to study functional aspects of an organism, the delivered radiation dose to the patient, and the achievable resolution. In the last few decades, spectroscopists and imaging scientists have been interested in the use of terahertz (THz) frequencies (30 µm to 3 mm wavelength) due to the low photon energy associated (E∼1 meV, not causing breaking of the molecular bonds but still interacting with some vibrational modes) and the high penetration depth that is achievable. THz has been already adopted in security, quality control and material sciences. However, the adoption of THz frequencies for biological and clinical imaging means to face, as a major limitation, the very scarce resolution associated with the use of such long wavelengths. To address this aspect and reconcile the benefit of minimal harmfulness for bioimaging with the achievable resolving power, many attempts have been made. This review summarises the state-of-the-art of THz imaging applications aimed at achieving super-resolution, describing how practical aspects of optics and quasi-optics may be treated to efficaciously implement the use of THz as a new low-dose and versatile modality in biomedical imaging and clinical research.

Revealing inscriptions obscured by time on an early-modern lead funerary cross using terahertz multispectral imaging.

The presence of a corrosion layer on lead art and archæological objects can severely impede the interpretation of inscriptions, thus hampering our overall understanding of the object and its context. While the oxidation of lead that dominates corrosion may be chemically reversible via reduction, potentially providing some access to inscriptions otherwise obscured by time, corrosion damage is overall neither entirely reversible nor is the reduction process in all cases easy or feasible to carry out. In this study, by taking advantage of the unique penetration ability of terahertz radiation and the abundant frequency bands covered by a single-cycle terahertz pulse, we perform nondestructive terahertz multispectral imaging to look under the corrosion on a sixteenth century lead funerary cross (croix d'absolution) from Remiremont in Lorraine, France. The multispectral images obtained from various terahertz frequency bands are fed into a judiciously designed post-processing chain for image restoration and enhancement, thus allowing us for the first time to read obscured inscriptions that might have otherwise been lost. Our approach, which brings together in a new way the THz properties of the constituent materials and advanced signal- and image-processing techniques, opens up new perspectives for multi-resolution analysis at terahertz frequencies as a technique in archæometry and will ultimately provide unprecedented information for digital acquisition and documentation, character extraction, classification, and recognition in archæological studies.

Visualising liquid transport through coated pharmaceutical tablets using Terahertz pulsed imaging.

Dissolution of pharmaceutical tablets is a complex process, especially for coated tablets where layered structures form an additional barrier for liquid transport into the porous tablet matrix. A better understanding of the role of the coating structure in the mass transport processes that govern drug release, starting with the wetting of the coating layer by the dissolution medium, can benefit the formulation design and optimisation of the production. For this study, terahertz pulsed imaging was used to investigate how dissolution medium can penetrate coated tablets. In order to focus on the fundamental process, the model system for this proof-of-principle study consisted of tablet cores made from pure microcrystalline cellulose compacted to a defined porosity coated with Opadry II, a PVA-based immediate release coating blend. The coating was applied to a single side of flat-faced tablets using vacuum compression moulding. It was possible to resolve the hydration of the coating layer and the subsequent liquid ingress into the dry tablet core. The analysis revealed a discontinuity in density at the interface between coating and core, where coating polymer could enter the pore space at the immediate surface of the tablet cores during the coating process. This structure affected the liquid transport of the dissolution medium into the core. We found evidence for the formation of a gel layer upon hydration of the coating polymer. The porosity of the tablet core impacted the quality of coating and thus affected its dissolution performance (r =  0.6932 for the effective liquid penetration rate RPeff and the core porosity). This study established a methodology and can facilitate a more in-depth understanding of the role of coating on tablet dissolution.

Terahertz imaging demonstrates its diagnostic potential and reveals a relationship between cutaneous dehydration and neuropathy for diabetic foot syndrome patients.

Diabetic foot syndrome, a long term consequence of Diabetes Mellitus, is the most common cause of non-traumatic amputations. Around 8% of the world population suffers from diabetes, 15% of diabetic patients present a diabetic foot ulcer which leads to amputation in 2.5% of the cases. There is no objective method for the early diagnosis and prevention of the syndrome and its consequences. We test terahertz imaging, which is capable of mapping the cutaneous hydration, for the evaluation of the diabetic foot deterioration as an early diagnostic test as well as ulcers prevention and tracking tool. Furthermore, the analysis of our terahertz measurements combined with neurological and vascular assessment of the patients indicates that the dehydration is mainly related to the peripheral neuropathy without a significant vascular cause.

THz medical imaging: from in vitro to in vivo.

Terahertz (THz) radiation has attracted considerable attention in medical imaging owing to its nonionizing and spectral fingerprinting characteristics. To date, most studies have focused on in vitro and ex vivo objects with water-removing pretreatment because the water in vivo excessively absorbs the THz waves, which causes deterioration of the image quality. In this review, we discuss how THz medical imaging can be used for a living body. The development of imaging contrast agents has been particularly useful to this end. In addition, we also introduce progress in novel THz imaging methods that could be more suitable for in vivo applications. Based on our discussions, we chart a developmental roadmap to take THz medical imaging from in vitro to in vivo.

Development of a mechanism for reconstruction of terahertz single-frequency images of biological samples.

Algorithmic mechanisms are used to improve terahertz (THz) image quality, which is critical to a biological sample analysis. A complete mechanism for the super-resolution reconstruction and evaluation of THz biological sample images was constructed in this study. With eucalyptus leaves as an example, the THz spectral region screening technique was adopted to select the characteristic frequencies for imaging, and the THz single-frequency images were reconstructed with the single-image super-resolution image reconstruction technique. The THz super-resolution reconstructed images without ideal reference were evaluated after the introduction of three no-reference image evaluation criteria considering the diversity and complexity of organisms. The results show that the THz image reconstruction mechanism proposed in this study led to an increase in resolution and a decrease in noise. At the same time, the imaging quality of biological samples was considerably improved, and the detailed information was enriched. These provide a reference for a THz imaging analysis of leaves and other biological samples.

Terahertz sensor based on a three-dimensional double I-type metamaterial integrated microfluidic channel.

Terahertz metamaterial sensors have received extensive attention in biosensing applications. However, sensitivity toward terahertz frequencies emitted by liquid samples remains challenging because of the strong absorption of terahertz waves by water. Here, we present a highly sensitive terahertz sensor based on a three-dimensional double I-type metamaterial integrated microfluidic channel. The designed sensor produces an inductive-capacitive (LC) resonance with a high quality factor of approximately 72, while demonstrating a maximum sensitivity of 832 GHz/RIU. Furthermore, we analyzed the relationship between the resonance frequency and ethanol concentration. These findings would promote the application of terahertz technology in label-free and rapid biomedical sensing as well as substance detection.

Trace Identification and Visualization of Multiple Benzimidazole Pesticide Residues on Toona sinensis Leaves Using Terahertz Imaging Combined with Deep Learning.

Molecular spectroscopy has been widely used to identify pesticides. The main limitation of this approach is the difficulty of identifying pesticides with similar molecular structures. When these pesticide residues are in trace and mixed states in plants, it poses great challenges for practical identification. This study proposed a state-of-the-art method for the rapid identification of trace (10 mg·L-1) and multiple similar benzimidazole pesticide residues on the surface of Toona sinensis leaves, mainly including benzoyl (BNL), carbendazim (BCM), thiabendazole (TBZ), and their mixtures. The new method combines high-throughput terahertz (THz) imaging technology with a deep learning framework. To further improve the model reliability beyond the THz fingerprint peaks (BNL: 0.70, 1.07, 2.20 THz; BCM: 1.16, 1.35, 2.32 THz; TBZ: 0.92, 1.24, 1.66, 1.95, 2.58 THz), we extracted the absorption spectra in frequencies of 0.2-2.2 THz from images as the input to the deep convolution neural network (DCNN). Compared with fuzzy Sammon clustering and four back-propagation neural network (BPNN) models (TrainCGB, TrainCGF, TrainCGP, and TrainRP), DCNN achieved the highest prediction accuracies of 100%, 94.51%, 96.26%, 94.64%, 98.81%, 94.90%, 96.17%, and 96.99% for the control check group, BNL, BCM, TBZ, BNL + BCM, BNL + TBZ, BCM + TBZ, and BNL + BCM + TBZ, respectively. Taking advantage of THz imaging and DCNN, the image visualization of pesticide distribution and residue types on leaves was realized simultaneously. The results demonstrated that THz imaging and deep learning can be potentially adopted for rapid-sensing detection of trace multi-residues on leaf surfaces, which is of great significance for agriculture and food safety.

Near-field Terahertz Sensing of HeLa Cells and Pseudomonas Based on Monolithic Integrated Metamaterials with a Spintronic Terahertz Emitter.

Label-free biosensors operating within the terahertz (THz) spectra have helped to unlock a myriad of potential THz applications, ranging from biomaterial detection to point-of-care diagnostics. However, the THz wave diffraction limit and the lack of emitter-integrated THz biosensors hinder the proliferation of high-resolution near-field label-free THz biosensing. Here, a monolithic THz emission biosensor (TEB) is achieved for the first time by integrating asymmetric double-split ring resonator metamaterials with a ferromagnetic heterojunction spintronic THz emitter. This device exhibits an electromagnetically induced transparency window with a resonance frequency of 1.02 THz and a spintronic THz radiation source with a bandwidth of 900 GHz, which are integrated on a fused silica substrate monolithically for the first time. It was observed that the resonance frequency experienced a red-shift behavior with increasing concentration of HeLa cells and Pseudomonas because of the strong interaction between the spintronic THz radiation and the biological samples on the metamaterials. The spatial frequency red-shift resolution is ∼0.01 THz with a Pseudomonas concentration increase from ∼0.5 × 104 to ∼1 × 104/mL. The monolithic THz biosensor is also sensitive to the sample concentration distribution with a 15.68 sensitivity under a spatial resolution of 500 µm, which is determined by the infrared pump light diffraction limit. This TEB shows great potential for high-resolution near-field biosensing applications of trace biological samples.

Terahertz Imaging and Characterization Protocol for Freshly Excised Breast Cancer Tumors.

This manuscript presents a protocol to handle, characterize, and image freshly excised human breast tumors using pulsed terahertz imaging and spectroscopy techniques. The protocol involves terahertz transmission mode at normal incidence and terahertz reflection mode at an oblique angle of 30°. The collected experimental data represent time domain pulses of the electric field. The terahertz electric field signal transmitted through a fixed point on the excised tissue is processed, through an analytical model, to extract the refractive index and absorption coefficient of the tissue. Utilizing a stepper motor scanner, the terahertz emitted pulse is reflected from each pixel on the tumor providing a planar image of different tissue regions. The image can be presented in time or frequency domain. Furthermore, the extracted data of the refractive index and absorption coefficient at each pixel are utilized to provide a tomographic terahertz image of the tumor. The protocol demonstrates clear differentiation between cancerous and healthy tissues. On the other hand, not adhering to the protocol can result in noisy or inaccurate images due to the presence of air bubbles and fluid remains on the tumor surface. The protocol provides a method for surgical margins assessment of breast tumors.

A Nondestructive Eggshell Thickness Measurement Technique Using Terahertz Waves.

Eggshells play a number of important roles in the avian and reptile kingdom: protection of internal contents and as a major source of minerals for developing embryos. However, when researching these respective roles, eggshell thickness measurement remains a bottleneck due to the lack of a non-destructive measurement techniques. As a result, many avian and reptile research protocols omit consideration of eggshell thickness bias on egg or embryo growth and development. Here, we validate a non-destructive method to estimate eggshell thickness based on terahertz (THz) reflectance spectroscopy using chicken white coloured eggs. Since terahertz waves are reflected from outer air-eggshell interface, as well as the inner eggshell-membrane boundary, the resulting interference signals depend on eggshell thickness. Thus, it is possible to estimate shell thickness from the oscillation distance in frequency-domain. A linear regression-based prediction model for non-destructive eggshell thickness measurement was developed, which had a coefficient of determination (R2) of 0.93, RMSEP of 0.009, RPD of 3.45 and RER 13.67. This model can estimate eggshell thickness to a resolution of less than 10 µm. This method has the potential to expand the protocols in the field of avian and reptile research, as well as be applied to industrial grading of eggs.

Three-dimensional water mapping of succulent Agave victoriae-reginae leaves by terahertz imaging.

While terahertz imaging has been used before for the determination of water content in vegetative tissue, most studies have either presented measurements of the temporal evolution of water content at a single-point of the plant or have presented two-dimensional images of leaves, demonstrating the potential of the technique, but relatively little of such information has been used to support biologically relevant conclusions. In this article we introduce terahertz time-domain spectroscopic imaging as a technique for the determination of the three-dimensional distribution of water in succulent plant tissues. We present the first three-dimensional water mapping of an agave leaf, which demonstrates an unprecedented capability to study the water retention mechanisms within succulent plants. We found that agave leaves are composed of a low-hydration outer tissue layer, defined by the outermost layer of vascular tissue that surrounds a high-hydration tissue, the carbohydrate rich hydrenchyma. The findings are supported by histological images and the correlation between the water content and carbohydrate presence is consistent with recently published findings of a remarkably large hydration shell associated with agave fructans.

At-line validation of optical coherence tomography as in-line/at-line coating thickness measurement method.

Optical Coherence Tomography (OCT) is a promising technology for monitoring of pharmaceutical coating processes. However, the pharmaceutical development and manufacturing require a periodic validation of the sensor's accuracy. For this purpose, we propose polyethylene terephthalate (PET) films as a model system, to periodically validate the measurements during manufacturing. This study proposes a new approach addressing the method validation requirement in the pharmaceutical industry and presents results for complementary methods. The methods investigated include direct measurement of the layer thickness using a micrometer gauge as reference, X-ray micro computed tomography, transmission and reflectance terahertz pulsed imaging, as well as 1D- and 3D-OCT. To quantify the significance of OCT for pharmaceutical coatings, we compared the OCT results for commercial Thrombo ASS and Pantoloc tablets with direct measurements of coating thickness via light microscopy of microtome cuts. The results of both methods correlate very well, indicating high intra- and inter-tablet variations in the coating thickness for the commercial tablets. The light microscopy average measured coating thickness of Thrombo ASS (Pantoloc) was 71.0 µm (83.7 µm), with an inter-coating variability of 8.7 µm (6.5 µm) and an intra-coating variability of 2.3 µm to 9.4 µm (2.1 µm to 6.7 µm).

Determining the effect of photodegradation on film coated nifedipine tablets with terahertz based coating thickness measurements.

Film coating of nifedipine tablets is commonly performed to reduce photo-degradation. The coating thickness of these tablets is a primary dictating factor of photo-stability. Terahertz spectroscopy enables accurate measurement of coating thickness. This study identifies a method to determine an end-point of a photo-protective coating process by using coating thickness measurements from terahertz time of flight spectroscopy (THz-TOF). For this method, nifedipine tablets, at different coating thicknesses, were placed in a photostability chamber. The illumination conditions of the coated tablets were adjusted based on the time duration of these tablets inside the chamber. A multiple linear regression model was developed with the coating thickness estimates from THz-TOF and illumination conditions information to predict the amount of drug remaining after photo-degradation (percent label claim). The prediction error of this model was 1.03% label claim in the range of 88.4-100.6% label claim. According to this model, acceptable levels of photo-protection in illumination conditions of up to approximately 700,000 lx hours was achieved at the end of the coating process (approximately 50 µm coating thickness) performed in this study. These results suggest THz-TOF as a viable process analytical technology tool for process understanding and end-point determination of a photo-protective coating process.