Search

With the outbreak and pandemic of COVID-19, point-of-care testing (POCT) systems have been attracted much attention due to their significant advantages of small batches of samples, user-friendliness, easy-to-use and simple detection. Among them, flexible biosensors show practical significance as their outstanding properties in terms of flexibility, portability, and high efficiency, which provide great convenience for users. To construct highly functional flexible biosensors, abundant kinds of polymers substrates have been modified with sufficient properties to address certain needs. Paper-based biosensors gain considerable attention as well, owing to their foldability, lightweight and adaptability. The other important flexible biosensor employs textiles as substrate materials, which has a promising prospect in the area of intelligent wearable devices. In this feature article, we performed a comprehensive review about the applications of flexible biosensors based on the classification of substrate materials (polymers, paper and textiles), and illustrated the strategies to design effective and artificial sensing platforms, including colorimetry, fluorescence, and electrochemistry. It is demonstrated that flexible biosensors play a prominent role in medical diagnosis, prognosis, and healthcare.

Significance: Histopathological analysis of tissues is an essential tool for grading, staging, diagnosing and resecting cancers and other malignancies. Current histopathological techniques require substantial sample processing prior to staining with hematoxylin and eosin (H&E) dyes, to highlight nuclear and cellular morphology. Sample preparation and staining is resource-intensive and introduces potential for variability during sample preparation. Aim: We present a novel method for direct label-free histopathological assessment of formalin fixed paraffin embedded tissue blocks and thin tissue sections using a dual contrast photoacoustic remote sensing (PARS) microscopy system. Approach: To emulate the nuclear and cellular contrast of H&E staining, we leverage unique properties of the PARS system. Here the ultraviolet excitation of the PAARS microscope takes advantage of DNA's unique optical absorption to provide nuclear contrast analogous to hematoxylin staining of cell nucelli. Concurrently, the optical scattering contrast of the PARS detection system is leveraged to provide bulk tissue contrast analogous to eosin staining of cell membranes. Results: We demonstrate the efficacy of this technique by imaging human breast tissue and human skin tissues in formalin fixed paraffin embedded tissue blocks and frozen sections respectively. Salient nuclear and extra-nuclear features such as cancerous cells, glands and ducts, adipocytes, and stromal structures such as collagen. Conclusions. The presented dual contrast PARS microscope enables label-free visualizations of tissue with contrast and quality analogous to the current gold standard for histopathological analysis. Thus, the proposed system is well positioned to augment existing histopathological workflows, providing histological imaging directly on unstained tissue blocks and sections., 13 Pages, 3 figures

Introduction: Alternative methods of dry storage and transportation may be a viable alternative to the use of liquid storage medium for cervical samples, especially for screening programs in places with few resources. Objective: The objective of this study is to verify the viability and efficacy of human papillomavirus DNA (HPV-DNA) detection in cervical cell samples collected and stored on a Flinders Technology Associates (FTA) card (Whatman Indicating FTA® Elute Micro Card) and subsequently recovered in ethanol-based liquid medium and to compare the results to those obtained using samples stored directly in ethanol-based liquid medium. Study Design: Thirty-four women submitted to ETZ (excision of the transformation zone of the cervix) were included in this study. Before ETZ, 2 samples of exfoliated cervical cells were collected from each woman by a doctor and stored in ethanol-based liquid medium and on an FTA card. DNA recovery from FTA samples was performed using ethanol-based liquid medium. Detection of HPV-DNA in the samples was performed using the Cobas® 4800 HPV Test Platform. Results and Conclusions: The HPV-DNA detection positivity rates were 70.6% for the samples collected directly in liquid medium and 64.7% for the samples stored on the FTA card, with high detection accuracy in the DNA samples recovered from the FTA card (area under the curve = 0.958; 95% confidence interval = 0.890–1.000). The concordance between the results obtained using the 2 storage media was 94.1% (Kappa = 0.866). These preliminary results suggest that collection of cervical material on an FTA card may be an alternative to storage in liquid medium since the liquid medium has some limitations. In addition, DNA recovery from the card using ethanol-based liquid medium streamlines the workflow in the laboratory and reduces the cost associated with reagents, thereby facilitating access to the HPV test in places with few resources and potentially improving cervical cancer screening.

Significance: To advance our understanding of the contrast observed when imaging with polarization-sensitive optical coherence tomography (PS-OCT) and its correlation with oral cancerous pathologies, a detailed comparison with histology provided via ex vivo fixed tissue is required. The effects of tissue fixation, however, on such polarization-based contrast have not yet been investigated. Aim: A study was performed to assess the impact of tissue fixation on depth-resolved (i.e., local) birefringence measured with PS-OCT. Approach: A PS-OCT system based on depth-encoded polarization multiplexing and polarization-diverse detection was used to measure the Jones matrix of a sample. A wide variety of ex vivo samples were measured freshly after excision and 24 h after fixation, consistent with standard pathology. Some samples were also measured 48 h after fixation. Results: The tissue fixation does not diminish the birefringence contrast. Statistically significant changes were observed in 11 out of 12 samples; these changes represented an increase in contrast, overall, by 11% on average. Conclusions: We conclude that the fixed samples are suitable for studies seeking a deeper understanding of birefringence contrast in oral tissue pathology. The enhancement of contrast removes the need to image immediately postexcision and will facilitate future investigations with PS-OCT and other advanced polarization-sensitive microscopy methods, such as mapping of the local optic axis with PS-OCT and PS-optical coherence microscopy.

2Naphthalene sulfonate (2NS) is an intermediate compound used in textile industries. Being nonbiodegradable, the concerns regarding its biotoxicity have risen. In the present investigation the toxic effects of 2NS were analyzed with the help of Attenuated Total Reflection Fourier Transform Infrared (ATR-FTIR), which was used to monitor changes in the vibrational modes of functional groups within the biomolecules. After calculating LD 50, one half of LD 50 i.e. 0.33 mg/15 g b.w. was intraperitoneally administrated and the brain tissue was collected for investigation after 96 h of exposure. The spectra observed revealed the significant differences in absorbance and areas between control and treated groups reflecting the change in proteins, lipids and nucleic acid due to toxicity induced by 2NS. In addition, protein secondary structure analysis was focused in this study, which reveals alterations in α helix and β sheet structure after 2NS intoxication. Histopathology of brain was also studied, which reveals changes in the histology of brain in group treated with 2NS. In conclusion, the study highlighted the application of ATR-FTIR and histopathology for toxicity assessment.

Significance: Processing and diagnosing a set of 12 prostate biopsies using conventional histology methods typically take at least one day. A rapid and accurate process performed while the patient is still on-site could significantly improve the patient’s quality of life. Aim: We develop and assess the feasibility of a one-hour-to-diagnosis (1Hr2Dx) method for processing and providing a preliminary diagnosis of a set of 12 prostate biopsies. Approach: We developed a fluorescence staining, optical clearing, and 3D open-top light-sheet microscopy workflow to enable 12 prostate needle core biopsies to be processed and diagnosed within an hour of receipt. We analyzed 44 biopsies by the 1Hr2Dx method, which does not consume tissue. The biopsies were then processed for routine, slide-based 2D histology. Three pathologists independently evaluated the 3D 1Hr2Dx and 2D slide-based datasets in a blinded, randomized fashion. Turnaround times were recorded, and the accuracy of our method was compared with gold-standard slide-based histology. Results: The average turnaround time for tissue processing, imaging, and diagnosis was 44.5 min. The sensitivity and specificity of 1Hr2Dx in diagnosing cancer were both >90%. Conclusions: The 1Hr2Dx method has the potential to improve patient care by providing an accurate preliminary diagnosis within an hour of biopsy.

Significance: Research in tissue engineering and in vitro organ formation has recently intensified. To assess tissue morphology, the method of choice today is restricted primarily to histology. Thus novel tools are required to enable noninvasive, and preferably label-free, three-dimensional imaging that is more compatible with futuristic organ-on-a-chip models. Aim: We investigate the potential for using multiphoton microscopy (MPM) as a label-free in vitro approach to monitor calcium-induced epidermal differentiation. Approach: In vitro epidermis was cultured at the air–liquid interface in varying calcium concentrations. Morphology and tissue architecture were investigated using MPM based on visualizing cellular autofluorescence. Results: Distinct morphologies corresponding to epidermal differentiation were observed. In addition, Ca2+-induced effects could be distinguished based on the architectural differences in stratification in the tissue cultures. Conclusions: Our study shows that MPM based on cellular autofluorescence enables visualization of Ca2+-induced differentiation in epidermal skin models in vitro. The technique has potential to be further adapted as a noninvasive, label-free, and real-time tool to monitor tissue regeneration and organ formation in vitro.

Purpose: Prostate cancer primarily arises from the glandular epithelium. Histomophometric techniques have been used to assess the glandular epithelium in automated detection and classification pipelines; however, they are often rigid in their implementation, and their performance suffers on large datasets where variation in staining, imaging, and preparation is difficult to control. The purpose of this study is to quantify performance of a pixelwise segmentation algorithm that was trained using different combinations of weak and strong stroma, epithelium, and lumen labels in a prostate histology dataset. Approach: We have combined weakly labeled datasets generated using simple morphometric techniques and high-quality labeled datasets from human observers in prostate biopsy cores to train a convolutional neural network for use in whole mount prostate labeling pipelines. With trained networks, we characterize pixelwise segmentation of stromal, epithelium, and lumen (SEL) regions on both biopsy core and whole-mount H&E-stained tissue. Results: We provide evidence that by simply training a deep learning algorithm on weakly labeled data generated from rigid morphometric methods, we can improve the robustness of classification over the morphometric methods used to train the classifier. Conclusions: We show that not only does our approach of combining weak and strong labels for training the CNN improve qualitative SEL labeling within tissue but also the deep learning generated labels are superior for cancer classification in a higher-order algorithm over the morphometrically derived labels it was trained on.

Paracrine and endocrine roles have increasingly been ascribed to extracellular vesicles (EVs) generated by multicellular organisms. Central to the biogenesis, content, and function of EVs are their delimiting lipid bilayer membranes. To evaluate research progress on membranes and EVs, the International Society for Extracellular Vesicles (ISEV) conducted a workshop in March 2018 in Baltimore, Maryland, USA, bringing together key opinion leaders and hands-on researchers who were selected on the basis of submitted applications. The workshop was accompanied by two scientific surveys and covered four broad topics: EV biogenesis and release; EV uptake and fusion; technologies and strategies used to study EV membranes; and EV transfer and functional assays. In this ISEV position paper, we synthesize the results of the workshop and the related surveys to outline important outstanding questions about EV membranes and describe areas of consensus. The workshop discussions and survey responses reveal that while much progress has been made in the field, there are still several concepts that divide opinion. Good consensus exists in some areas, including particular aspects of EV biogenesis, uptake and downstream signalling. Areas with little to no consensus include EV storage and stability, as well as whether and how EVs fuse with target cells. Further research is needed in these key areas, as a better understanding of membrane biology will contribute substantially towards advancing the field of extracellular vesicles. Fil: Russell, Ashley E.. University Johns Hopkins; Estados Unidos Fil: Sneider, Alexandra. University Johns Hopkins; Estados Unidos Fil: Witwer, Kenneth W.. University Johns Hopkins; Estados Unidos Fil: Bergese, Paolo. Università Degli Studi Di Brescia; Italia Fil: Bhattacharyya, Suvendra N.. Indian Institute of Chemical Biology; India Fil: Cocks, Alexander. Cardiff University; Reino Unido Fil: Cocucci, Emanuele. Ohio State University; Estados Unidos Fil: Erdbrügger, Uta. University of Virginia; Estados Unidos Fil: Falcon Perez, Juan M.. Ikerbasque Basque Foundation for Science; España Fil: Freeman, David W.. National Institute On Aging National Institute for Helth ; Estados Unidos Fil: Gallagher, Thomas M.. Loyola University Of Chicago; Estados Unidos Fil: Hu, Shuaishuai. Technological University Dublin; Irlanda Fil: Huang, Yiyao. University Johns Hopkins; Estados Unidos. Southern Medical University; China Fil: Jay, Steven M.. University of Maryland; Estados Unidos Fil: Kano, Shin-ichi. The University of Alabama at Birmingham School of Medicine; Estados Unidos Fil: Lavieu, Gregory. Institut Curie; Francia Fil: Leszczynska, Aleksandra. University of California at San Diego; Estados Unidos Fil: Llorente, Alicia M.. Oslo University Hospital; Noruega Fil: Lu, Quan. Harvard University. Harvard School of Public Health; Estados Unidos Fil: Mahairaki, Vasiliki. University Johns Hopkins; Estados Unidos Fil: Muth, Dillon C.. University Johns Hopkins; Estados Unidos Fil: Noren Hooten, Nicole. National Institute On Aging National Institute for Helth ; Estados Unidos Fil: Ostrowski, Matias. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas en Retrovirus y Sida. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas en Retrovirus y Sida; Argentina Fil: Prada, Ilaria. Consiglio Nazionale delle Ricerche; Italia Fil: Sahoo, Susmita. Icahn School of Medicine at Mount Sinai ; Estados Unidos Fil: Schøyen, Tine Hiorth. Uit The Arctic University Of Norway; Noruega. University Johns Hopkins; Estados Unidos Fil: Sheng, Lifuy. University of Washington. School of Medicine; Estados Unidos Fil: Tesch, Deanna. Shaw University; Estados Unidos Fil: Van Niel, Guillaume. No especifíca; Fil: Vandenbroucke, Roosmarijn E.. University of Ghent; Bélgica Fil: Verweij, Frederik J.. No especifíca; Fil: Villar, Ana V.. Universidad de Cantabria; España Fil: Wauben, Marca. University of Utrecht; Países Bajos Fil: Wehman, Ann M.. Universität Würzburg; Alemania Fil: Ardavan, Arzhang. Peking University; ; China Fil: Carter, David Raul Francisco. Oxford Brookes University; Reino Unido Fil: Vader, Pieter. University Medical Center Utrecht; Países Bajos

Molecular testing of tumors from formalin-fixed paraffin-embedded (FFPE) tissue blocks is central to clinical practice; however, it requires histology support and increases test turnaround time. Prospective fresh frozen tissue collection requires special handling, additional storage space, and may not be feasible for small specimens. Filter paper-based collection of tumor DNA reduces the need for histology support, requires little storage space, and preserves high-quality nucleic acid. We investigated the performance of tumor smears on filter paper in solid tumor genotyping, as compared with paired FFPE samples. Whatman FTA Micro Card (FTA preps) smears were prepared from 21 fresh tumor samples. A corresponding cytology smear was used to assess tumor cellularity and necrosis. DNA was isolated from FTA preps and FFPE core samples using automated methods and quantified using SYBR green dsDNA detection. Samples were genotyped for 471 mutations on a mass spectrophotometry-based platform (Sequenom). DNA concentrations from FTA preps and FFPE correlated for untreated carcinomas but not for mesenchymal tumors (Spearman σ=0.39 and σ=-0.1, respectively). Average DNA concentrations were lower from FTA preps as compared with FFPE, but DNA quality was higher with less fragmentation. Seventy-six percent of FTA preps and 86% of FFPE samples generated adequate DNA for genotyping. FTA preps tended to perform poorly for collection of DNA from pretreated carcinomas and mesenchymal neoplasms. Of the 16 paired DNA samples that were genotyped, 15 (94%) gave entirely concordant results. Filter paper-based sample preservation is a feasible alternative to FFPE for use in automated, high-throughput genotyping of carcinomas.

Mueller microscopy studies of fixed unstained histological cuts of human skin models were combined with an analysis of experimental data within the framework of differential Mueller matrix (MM) formalism. A custom-built Mueller polarimetric microscope was used in transmission configuration for the optical measurements of skin tissue model adjacent cuts of various nominal thicknesses (5 to 30 mm). The maps of both depolarization and polarization parameters were calculated from the corresponding microscopic MM images by applying a logarithmic Mueller matrix decomposition (LMMD) pixelwise. The parameters derived from LMMD of measured tissue cuts and the intensity of transmitted light were used for an automated segmentation of microscopy images to delineate dermal and epidermal layers. The quadratic dependence of depolarization parameters and linear dependence of polarization parameters on thickness, as predicted by the theory, was confirmed in our measurements. These findings pave the way toward digital histology with polarized light by presenting the combination of optimal optical markers, which allows mitigating the impact of tissue cut thickness fluctuations and increases the contrast of polarimetric images for tissue diagnostics.

Intraoperative assessment of breast surgical margins will be of value for reducing the rate of re-excision surgeries for lumpectomy patients. While frozen-section histology is used for intraoperative guidance of certain cancers, it provides limited sampling of the margin surface (typically

This study aims to determine if light scatter parameters measured with spatial frequency domain imaging (SFDI) can accurately predict stromal, epithelial, and adipose fractions in freshly resected, unstained human breast specimens. An explicit model was developed to predict stromal, epithelial, and adipose fractions as a function of light scattering parameters, which was validated against a quantitative analysis of digitized histology slides for N = 31 specimens using leave-one-out cross-fold validation. Specimen mean stromal, epithelial, and adipose volume fractions predicted from light scattering parameters strongly correlated with those calculated from digitized histology slides (r = 0.90, 0.77, and 0.91, respectively, p-value