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Past, present and future: overview on Histology and histopathology
Abstract
First paragraph (this article has no abstract)
With sincere satisfaction and pride, I present this first editorial of the Journal of Histology and Histopathology edited by myself. I know this is a very challenging task, assuming the
... The histopathological examination of endometrial biopsy or full-thickness sections remains indispensable in assessing the alterations in the uterine wall structure [3,13,39,40]; hematoxylin and eosin (HE) is a routinely used staining method to visualize tissue morphology. The HE stain is commonly used to assess connective tissue fibers as well, besides the availability of well-established histochemical methods for the visualization of connective tissue and fibrosis [1,5,[41][42][43][44][45][46][47]. As the HE is not specific to connective tissue, the use of special stains is potentially beneficial, especially for the quantitative assessment of fibrosis. ...
... While some claim that specific staining does not provide any additional value to the breeding soundness exam [15,43], assessing the scale and severity of fibrosis is considered more reliable when a special stain is used. Moreover, in scientific research on the assessment of connective tissue in the endometrium, the specific staining for fibers is widely used and recommended [45][46][47][68][69][70][71]. This review serves to systematically summarize the usefulness of methods of connective tissue staining in the equine uterus, which make the diagnosis of endometrial and cervical diseases more accurate. ...
... Thus, staining selection should be directly related to the aim of the study and the possibility of evaluation. HE is globally used for the evaluation of cell morphology and tissue architecture [45]. It is commonly used to diagnose any histological alterations in a tissue, including fibrosis in endometrial samples, although it is not a specific stain for the connective tissue [68,69,87,88]. ...
Simple Summary
Uterine diseases are the leading cause of infertility in mares, causing increasing costs and losses in horses’ breeding. Their current diagnosis is often supported by obtaining endometrial biopsies and hematoxylin–eosin staining, which is the basic staining used in histopathology. This review aims to show the variety of uterine changes affecting fertility and highlights the usefulness of special stains for connective tissue visualization—Masson trichrome, picrosirius red, elastica van Gieson, or periodic acid–Schiff—for a more comprehensive diagnosis. The fibrosis evaluation includes connective tissue changes in the cervix, the endometrium, and around/in the wall of blood vessels. Cervical connective tissue undergoes cyclic changes impacting fertility, whereas vascular changes, especially in multiparous mares, are crucial for adapting to physiological shifts, affecting early pregnancy and hindering placental development. Special stains are valuable for the identification of structural changes in the cervix and fibrosis in uterine blood vessels. Moreover, equine endometrosis, linked to fibrotic processes in the endometrium, emphasizes the need for wider use of special stains in diagnosis. Therefore, we advocate for special staining in reproductive tract evaluation due to its simplicity, accessibility, and effectiveness. We encourage scientists and diagnosticians to adopt additional tools for clearer pathology visualization, enabling reliable fertility predictions.
Abstract
Uterine diseases stand as the primary cause of infertility in mares; however, the diagnostic process often relies on obtaining endometrial biopsies and their hematoxylin–eosin staining. This review seeks to present the variability of uterine changes and their impact on fertility and underscore the utility of special stains, such as Masson trichrome, picrosirius red, elastica van Gieson, or periodic acid–Schiff, in enhancing diagnostic breadth. Connective tissue evaluation in the cervix is discussed, as it is subjected to cyclic changes and the impact on overall fertility. Vascular changes, particularly prevalent in multiparous mares, play a crucial role in adapting to physiological and pathological alterations, affecting early gestation and impeding placental development. Given that uterine vascular pathologies often involve fibrotic changes, connective tissue stains emerge as a valuable tool in this context. Moreover, equine endometriosis, predominantly associated with endometrial fibrosis, further highlights the relevance of special stains, suggesting their underutilization in the diagnostic process. Recognizing the subjective nature of diagnosing uterine pathologies and the need for additional diagnostic tools, we advocate for using dedicated stains in the histopathological evaluation of uterine samples. In conclusion, we encourage scientists and diagnosticians to embrace additional tools that enhance pathology visualization, enabling more reliable diagnoses concerning expected fertility.
... These fixatives and staining agents paved the way for modern histological staining techniques, leading to the development of dyes still in use today (Black 2012). Notable examples include silver nitrate, which remains important in staining specific tissue structures, and trichrome stains, commonly applied in liver and kidney biopsies (Musumeci 2014). The invention of histologic stains, particularly aniline dyes in Germany in 1856, revolutionized the field by introducing numerous new staining techniques. ...
... A typical medical procedure for diagnosing tumors is staining, which involves applying a dye to the edges of sample tissues to identify diseased, tumorous, or other abnormal cells (Musumeci 2014). Staining is used in scientific investigations to identify cells and highlight proteins, nucleic acids, or analyze gel electrophoresis to facilitate microscopic examination (Jackson and Blythe 2013). ...
Improper husbandry techniques, environmental pollution, fluctuating weather conditions, and shifting market dynamics pose significant challenges to aquaculture and fisheries production, making fish more susceptible to diseases. Understanding and characterizing the interventions involved in disease outbreaks are crucial, as they can lead to significant financial losses in the aquaculture and fisheries sectors. Histopathology plays a vital role in fish disease diagnosis, as it involves the microscopic examination of tissue abnormalities. This study aimed to evaluate the tissue preparation procedures for aquatic organisms and provide a historical review of tissue staining techniques. Over time, histopathological staining methods have advanced, leading to the development of simpler, more precise, and more efficient techniques. While newer genetic, non-culture, immunostaining, and other advanced staining methods have largely replaced traditional staining procedures, several older techniques remain in use. To enhance their effectiveness, modern histological stains are being modified and combined with other staining methods. Additionally, emerging fields like proteomics can complement traditional approaches, such as pathogen identification and histological examination, to achieve more accurate disease diagnoses in aquaculture and fisheries.
... [4] In many circumstances, H&E stain may not be able to accurately identify or recognise the initial epithelial pathology that pathologists require, particularly in cases of early, micro-invasive squamous cell carcinoma (SCC), carcinoma in situ, and atypical epithelial malignancies. [5][6][7][8][9] Hence, diagnosing initial micro-invasiveness in OSCC using H&E stain alone will be challenging. In modern histology, the staining process is made more effective by combining various stains, which are then utilised to identify specific cells and structures. ...
... [10] A special or differential staining procedure that differentiates different tissue elements in a histology section can prove to be an easy and affordable fix facilitating an easy and accurate diagnosis of some challenging cases. [9,11] One such good differential stain capable of identifying epithelial cells in connective tissue stroma is the modified Cajal's trichrome stain (MCTS), which was developed by Gallego in 1919 after being first presented by Ramon Y. Cajal in 1897. It provides distinct hues in SCC according to the degree of cellular differentiation and keratinisation. ...
Background
Modified Cajal’s trichrome stain (MCTS) is a good differential stain that allows one to visibly distinguish between connective tissue and epithelial elements with different tonalities of colour.
Aim
Our study aims to evaluate and analyse the effectiveness of oral squamous cell carcinoma (OSCC) using MCTS.
Materials and Methods
A study was conducted retrospectively with 30 tissue blocks embedded in paraffin from cases of OSCC that have been confirmed by histopathology. Both standard haematoxylin and eosin (H&E) and MCTS were applied to each section. Then all the sections were analysed by two observers for nucleus cytoplasmic intensity, break in the basement membrane, and advancing front of the tumour, muscle, and surrounding stroma. The efficacy of the stain was assessed and was graded as 1, poor; 2, fair; and 3, good based on the staining intensity.
Statistical Analysis
The parameters were graded for H&E and modified Cajal’s stain. The results were subjected to the Chi-square test.
Result
The above-mentioned parameters analysed showed a uniformly significant P value of 0.001 for comparing modified Cajal’s trichrome stain to H&E stain. Measurement of the agreement was done based on Kappa statistics between two observers, and the values for each expression show that there was good agreement between the two for all the parameters.
Conclusion
MCTS can also be used as a diagnostic aid to pathologists for better distinction of cellular components and easier identification, thereby solving difficulties in diagnosis at earlier stages.
... Hematoxylin and Eosin staining is not always sufficient because not all substance features can be visualized. Therefore, special stains with higher substance selectivity are valuable tools in research practice [3,9,28]. ...
... An overview of the examined tissue enables collecting information on the visible macroscopic variance or the topography using the "naked eye." As for microscopic studies, light or electron microscopes are used to extract information about collected samples [28]. Microscopic examination precedes morphological gross examination to obtain quality samples for further tests. ...
Background: In anatomical research, there is often a need to present complex topographical relationships in more detail, under magnification, beyond the scope of gross anatomy. However, more than magnification is often needed to better understand the arrangement of small anatomical structures. Special stains allow for the differentiation of connective tissue stroma and mutual relations between small structures such as blood vessels and nerves. Aim: This review presents selected histochemical methods that can be applied in anatomical research. Such a concise outline can be both a compendium of knowledge for beginning researchers and an inspiration for anatomists and clinicians exploring the human body's details. Conclusions: The advantage of histochemical staining is its relatively low cost and the large amount of information that can be obtained with their use.
... [4] H and E stain is a permanent histologic stain which imparts blue color to the nuclei of the cells and stains the cytoplasm as pink. [5] The final diagnosis of some initial epithelial neoplasms may be challenging and difficult even with routine H and E staining, especially in cases of microinvasive oral squamous cell carcinoma (OSCC) and some atypical epithelial neoplasms. [6] In few cases, H and E stain may not provide the accurate identification or recognition of some abnormal components which the pathologists or researchers need. ...
... They are simple to interpret for diagnosis and remain as an essential auxiliary tool for routine histopathologic diagnosis. [5] One such It delineates various components in the tissue visually with different hues of color. This differential stain helps in the detection of tumors where connective tissue stromal components are associated with epithelial cells, for example, OSCC. ...
Introduction
In histopathologic laboratories, hematoxylin and eosin (H and E) staining is the routine and gold standard technique employed for biopsy tissues. But, few differential stains are being used in questionable diagnostic cases of oral squamous cell carcinoma (OSCC) to confirm its final diagnosis. Modified Cajal trichrome stain (CTS) is one such stain which may be employed as an adjunctive supporting aid for arriving at a conclusive diagnosis. To assess epithelial and connective tissue components in normal oral mucosa, oral epithelial dysplasia (OED), and OSCC cases stained with routine H and E and modified CTS. And also to compare and evaluate the efficacy of Modified CTS with that of H and E stain.
Materials and Methods
A total of 40 tissue samples, 10 each of normal oral mucosa and OED, 20 cases of OSCC (10 cases each of microinvasive and frank OSCC) were stained with standard H and E and modified CTS simultaneously.
Results
Modified CTS showed statistically significant and better results in differentiated terms of parameters such as depth of tumor invasion and differentiation between epithelial and connective tissue components in OSCC cases. Epithelial basement membrane interface and microinvasion are clearly appreciated in challenging cases of early epithelial malignancies such as microinvasive OSCC using this stain.
Conclusion
As this stain can easily delineate epithelial structures from various connective tissue components, it may be utilized as an auxiliary diagnostic support along with routine H and E stain for confirmatory diagnosis.
... Inside; water residues, molten salts, nutrients, etc. [1]. In biological studies, staining is used to mark nucleic acids, proteins, or gel electrophoresis to mark cells and aid microscopic examination [2]. ...
... When we look at the embedding process, the obtained tissues must be embeded before painting. The embedding process is made by using paraffin wax, which is used to increase the extraction [1]. However, as these fixatives are very useful, they can also have side effects, especially cell deterioration due to long-term exposure to the structures of the tissue. ...
... This has created a mindset in most life sciences researchers that only a pathologist can obtain information from their research slides. H& E is not only basic for clinical investigation, but is almost exclusively used in basic research [3]. ...
Basic and accurate histological examination of hematoxylin and eosin (H&E) stained slides is a critical skill for biomedical researchers. However, many postgraduate students struggle to interpret these slides, relying on external assistance for histological reporting. This knowledge gap can have long-term implications for research validity and career advancement. This paper presents a concise and structured approach to documenting H&E-stained histology slides, outlining essential assessment criteria and addressing common challenges. Emphasizing the importance of practice and experience in achieving proficiency, this framework aims to enhance research competency in biomedicine and promote accurate histological reporting.
... Histopathological refers to studies at the micro level caused by disease or abnormal biological tissues and cells changes. It has a crucial function in the identification and comprehension of different clinical situations, particularly in the diagnosis of cancer [1]. By conducting thorough analysis of specimens, histopathologists can discern the structural modifications that diseases cause in organs and tissues, offering crucial understanding of the characteristics, advancement, and possible therapeutic approaches for these conditions. ...
Sarcoidosis is a complex inflammatory disease involving many systems and its exact cause is still unknown. The characteristic in histopathology of it is the occurrence of epithelioid cell granulomas. The most common type of lymph node involvement is the lungs and chest has risks of other organs. Common diagnostic methods rely on clinical symptoms, imaging, and histopathology. Histopathology is the microscopic study of tissues affected by a disease and is essential for diagnosing sarcoidosis and differentiating it from other granulomatous disorders. According to different affected organs, the histologic pattern of sarcoidosis is also different. Lungs is the most affected organ. Granulomas are usually found within the interstitium, often with varying degrees of fibrosis. Cutaneous manifestations include a range of lesions such as erythema nodosum and lupus chilblain, and histological test may present dermal granulomas and lymphocytic infiltrates. Although liver involvement usually asymptomatic, its characteristics is a portal with a triple granuloma, occasionally accompanied by mild portal inflammation. This paper aims to investigate the histopathological findings of sarcoidosis in different organs.
... [11][12][13][14][15][16][17] Virtual staining circumvents the need for laborious sample preparation, specialized laboratory equipment, and technical personnel requirement in traditional chemical histological staining. 1,18 By leveraging deep learning for digital histological staining generation, virtual staining offers an efficient, precise, and cost-effective alternative. Deep learning-based virtual staining methods are primarily categorized into supervised and unsupervised approaches. ...
The emergence of virtual staining technology provides a rapid and efficient alternative for researchers in tissue pathology. It enables the utilization of unlabeled microscopic samples to generate virtual replicas of chemically stained histological slices, or facilitate the transformation of one staining type into another. The remarkable performance of generative networks, such as CycleGAN, offers an unsupervised learning approach for virtual coloring, overcoming the limitations of high-quality paired data required in supervised learning. Nevertheless, large-scale color transformation necessitates processing large field-of-view images in patches, often resulting in significant boundary inconsistency and artifacts. Additionally, the transformation between different colorized modalities typically needs further efforts to modify loss functions and tune hyperparameters for independent training of networks. In this study, we introduce a general virtual staining framework that is adaptable to various conditions. We propose a loss function based on the value mapping constraint to ensure the accuracy of virtual coloring between different pathological modalities, termed the Value Mapping Generative Adversarial Network (VM-GAN). Meanwhile, we present a confidence-based tiling method to address the challenge of boundary inconsistency arising from patch-wise processing. Experimental results on diverse data with varying staining protocols demonstrate that our method achieves superior quantitative indicators and improved visual perception.
... 4 In the discrimination of tumourous cells from the normal one, staining is performed at the rear and front ends of the tissue sample. 5 Staining is also performed to identify the cell structures, nucleic acid and proteins during microscopic investigation. 6 Multiple staining methods are employed to obtain an extensive diagnostic information like identifying the collagen, microorganisms, mucin etc in addition to tissue architecture simultaneously improving the accuracy of diagnosis. ...
Histopathology images of the lung biopsy tissues is considered as the gold standard modality to confirm the presence and
type of cancer. However, owing to the complex morphology, texture and presence of artifacts during sample preparation,
staining and imaging makes histopathology interpretation a challenging task and is subjective relying on the expertise of
pathologists. Additionally, the interpretation is laborious due to careful examination of multiples slides and multiple
regions within the slide adding burden to pathologists. Computer aided diagnosis from histopathology slides will greatly
assist the pathologists in their routine and will reduce the inter pathologist interpretation variability. An empirical study is
proposed to investigate the proficiency of three prominent pre-trained networks -AlexNet, GoogLeNet and ResNet-50
in classifying the lung histopathology images belonging to three categories of lung pathology, lung adenocarcinoma, lung
benign tissue and lung squamous cell carcinoma. The empirical study resulted in a phenomenal classification performance
by all three classifiers with ResNet-50 model taking a marginal advantage over the other two classifiers with a classification
accuracy of 99.26% and a Kappa coefficient of 0.989. AlexNet and GoogLeNet demonstrated a notable performance with
98.8% and 98.97% accuracy and a Kappa coefficient of 0.982 and 0.985.
... About 4-5-µm-thick sections were cut from all the paraffin blocks and stained with the H&E stain for evaluation of the histopathological changes. This included the assessment of granuloma formation and hepatic parenchymal changes (Drury andWallington 1980, Titford 2006;Musumeci 2014;Mossallam et al. 2015). ...
Despite the long history of experimental trials to combat schistosomiasis, it remains a significant burden due to drug resistance and the effectiveness of the standard treatment only against the mature stage, while skipping other early developmental stages thus leading to severe permanent pathological sequelae. Therefore, repurposing a commonly used well-known safe drug would be a wise alternative. We investigated the potential anti-schistosomal drug activity of Daflon® (DAF) against different schistosomal developmental stages. DAF was administrated at a dose of 100 mg/kg/mouse on days zero, 21, and 42 post-infection towards the invasive, immature, and mature stages of Schistosoma mansoni respectively in comparison to the standard anti-schistosomal drug (Praziquantel). All mice were sacrificed on day 49 post-infection. DAF induced a significant reduction in the total and female worm count, hepatic granuloma size, and number, the extent of liver parenchymal injury and fibrosis as well as intestinal and hepatic egg count compared to the infected untreated control. Liver malondialdehyde (MDA) levels significantly decreased in all DAF-treated groups. Scanning electron microscope findings revealed edema, tegumental blebs, cracks, and fissures in male tegument in all DAF-treated groups with distortion of the ventral suckers and disarrangement of the spines of the oral sucker. The female worm from DAF-treated groups showed tegumental edema with loss of the spines at the posterior end. Compared to the documented reduction of testosterone levels and distortion of testicular architecture in the S. mansoni-infected untreated group, DAF significantly restored testosterone levels and testicular architecture.
Graphical abstract
... To assess histopathological changes, specimens from the liver and the spleen of all infected GII subgroups were fixed in 10% of neutral formalin, sectioned and stained with H&E stain [24] . ...
... This information is crucial for accurate diagnosis, prognosis, and treatment planning in clinical practice. Histopathological studies will be conducted on various organs of fish, including the gills, intestine, liver, and muscle (Musumeci, 2014) ...
This book
compiles recent views, ideas, and advancements in the field of aquaculture pertaining to aquatic
animal health. It aims to cover all aspects of disease diagnostic techniques in aquaculture, covering
conventional methods, molecular biological techniques, and haemato-immunological techniques.
... Specimens from the liver and the spleen of the infected treated group and the infected non-treated control subgroup were fixed in 10% neutral formalin, sectioned, and stained with hematoxylin-eosin (H and E) stain to assess histologic alteration in different groups (Musumeci 2014). ...
Side effects and low efficacy of current anti-toxoplasmosis therapeutics against encysted bradyzoites necessitate research into alternative safe therapeutic options. The safety, immunostimulatory, and antimicrobial properties of alginate nanoparticle formulation (Alg-NP) highlight its potential as an oral therapy against acute toxoplasmosis. In the current study, Alg-NP was formulated and characterized and then assessed for its anti-Toxoplasma effects using parasitological, ultrastructural, immunological, and histopathological studies. Treatment with Alg-NP significantly prolonged mice survival and reduced the parasite burden in both peritoneal fluid and tissue impression smears. In addition, it altered parasite viability and caused severe tachyzoite deformities as evidenced by ultrastructural studies. Alg-NP induced high levels of serum IFN-γ in infected mice with significant amelioration in histopathological changes in both hepatic and splenic tissue sections. In conclusion, Alg-NP could be considered a promising therapeutic agent against acute murine toxoplasmosis, and owing to its safety, it could potentially be enlisted for human use.
... . At the microscopic level, histology and the field of cellular pathology, also referred to as histopathology, analyze patient tissues to understand the structural and functional alterations associated with diseases (Musumeci, 2014). While histopathology and the experimental laboratory techniques it routinely relies on are already being potentiated by rapid advances in computer science and artificial intelligence (AI) (de Matos et al., 2021), unique opportunities will be forthcoming to enhance cellular pathology as machine learning (ML) methods also extend the capabilities of cryogenic electron microscopy (cryoEM) and tomography (cryoET) and related techniques, facilitating the efficient, quantitative processing of patient derived imaging datasets. ...
Advances in cryogenic electron microscopy (cryoEM) single particle analysis have revolutionized structural biology by facilitating the in vitro determination of atomic- and near-atomic-resolution structures for fully hydrated macromolecular complexes exhibiting compositional and conformational heterogeneity across a wide range of sizes. Cryogenic electron tomography (cryoET) and subtomogram averaging are rapidly progressing toward delivering similar insights for macromolecular complexes in situ , without requiring tags or harsh biochemical purification. Furthermore, cryoET enables the visualization of cellular and tissue phenotypes directly at molecular, nanometric resolution without chemical fixation or staining artifacts. This forward-looking review covers recent developments in cryoEM/ET and related technologies such as cryogenic focused ion beam milling scanning electron microscopy and correlative light microscopy, increasingly enhanced and supported by artificial intelligence algorithms. Their potential application to emerging concepts is discussed, primarily the prospect of complementing medical histopathology analysis. Machine learning solutions are poised to address current challenges posed by “big data” in cryoET of tissues, cells, and macromolecules, offering the promise of enabling novel, quantitative insights into disease processes, which may translate into the clinic and lead to improved diagnostics and targeted therapeutics.
... Histomorphology provides information that cannot be obtained from external skeletal morphology (Bailleul et al., 2019;Kolb et al., 2015a;Quekett, 1849;Wang et al., 2020), such as ecological niches , sex differences via medullary bone (Bloom et al., 1941;Schweitzer et al., 2005Schweitzer et al., , 2007Wang et al., 2020), ontogenetic growth patterns (Erickson et al., 2004;Woodward, 2019;Woodward et al., 2014), pathological healing (Ekhtiari et al., 2020), and stress distribution in bones caused by animal locomotion (Bromage et al., 2003;de Margerie et al., 2004;Suniaga et al., 2018). Therefore, bone histomorphology has been emphasized not only in paleontological, but also in extant biological, pathological, and veterinary medical research (Musumci, 2014). ...
Observing bone microstructure may reveal aspects of the ecological behavior and physiological mechanisms of extant and extinct vertebrates. High quality bone thin sections are required for superior resolution of histomorphological observation. We encountered three problems in studying bone thin sections of the material available to us: (1) opacity due to post-mortem migration of blood pigment; (2) birefringence of the epoxy resin used to embed the samples which disturbs observations in a polarizing microscope; and (3) abrasive powder residue which becomes incorporated into the bone sections. Modified procedures were developed to solve these problems. The blood pigment can be removed by N,N,N’,N’-tetrakis (2-hydroxypropyl) ethylenediamine. The epoxy resin can be removed by repetitive treatments with acetone and dichloromethane. Water-proof silicon carbide papers can be used for grinding and polishing thin sections instead of abrasive powder. These improvements make clearer, more transparent sections with improved images, which are easier to measure.
... To gain a deep insight into pathologies and diseases, analyses at morphological, physiological, and functional levels are typically performed starting from histological slides [1]. For instance, in oncology, most tumour diagnoses go through some histology/histopathology analysis that is performed to describe and extract different features of the cancer cells which are illustrative of their in vivo behaviour [2]. Furthermore, in the era of immunotherapy, understanding the immune context beyond cancer cells is also pivotal for defining prognostic and predictive biomarkers [3]. ...
Most of the time, the deep analysis of a biological sample requires the acquisition of images at different time points, using different modalities and/or different stainings. This information gives morphological, functional, and physiological insights, but the acquired images must be aligned to be able to proceed with the co-localisation analysis. Practically speaking, according to Aristotle’s principle, “The whole is greater than the sum of its parts”, multi-modal image registration is a challenging task that involves fusing complementary signals. In the past few years, several methods for image registration have been described in the literature, but unfortunately, there is not one method that works for all applications. In addition, there is currently no user-friendly solution for aligning images that does not require any computer skills. In this work, DS4H Image Alignment (DS4H-IA), an open-source ImageJ/Fiji plugin for aligning multimodality, immunohistochemistry (IHC), and/or immunofluorescence (IF) 2D microscopy images, designed with the goal of being extremely easy to use, is described. All of the available solutions for aligning 2D microscopy images have also been revised. The DS4H-IA source code; standalone applications for MAC, Linux, and Windows; video tutorials; manual documentation; and sample datasets are publicly available.
... To assess histopathological changes, specimens from the liver and the spleen of all infected GII subgroups were fixed in 10% of neutral formalin, sectioned and stained with H&E stain [24] . ...
... Se sabe que Leeuwenhoek en el siglo XVIII, usó colorantes extraídos de plantas como el azafrán y el índigo solo para dar contraste a sus preparaciones, pero no hay evidencia de su uso en la observación de las células de la sangre solo en fibras musculares.3, 10,11,12 El principal componente natural del azafrán, responsable de producir color en solución es la crocetina, mientras que para el índigo es la indigotina (Figura 1). 13 15,16,17,18 La crocetina es una aglicona, presenta la estructura de un ácido conjugado poliinsaturado, cuatro grupos metilo y siete dobles enlaces conjugados. ...
... Poco a poco, a finales del siglo XIX y principios del XX, la interacción entre la histología y patología se hizo mucho más sólida, no se puede pensar en una sin la otra, y las dos requieren de la microtecnia. Gracias a esta herramienta se fundamentaron las bases de estas ciencias hasta la actualidad, y se cuenta con la madurez teórica y técnica para poder armar el complejo vital de la estructura viva (Musumeci, 2014). ...
A lo largo de la historia de nuestra civilización, los seres humanos hemos creado
una estrecha relación con los colores, como reflejo natural del mundo circundante.
Así como el hombre ancestral y el infante que se enfrentan por vez primera a la
comprensión de los colores primarios, el tono de sus artefactos más cercanos y los
matices de la naturaleza en los cambios de estación, esa relación evoluciona para
finalmente ver, observar y entender los tonos y los matices, además de crear mediante la aplicación del color (coloreando y tiñendo).
El planteamiento de la organización de los temas de este libro obedece principalmente a la necesidad que ha surgido dentro de los laboratorios de ciencias biológicas, a lo largo de los años, al hacer visible la materia orgánica, y revelar la delicada
estructura celular que permite que la vida exista.
Este libro posee diferentes cualidades para diferentes tipos de necesidades, desde
las del técnico laboratorista, quien necesita el número adecuado para solicitar la
compra del reactivo colorante; hasta las del estudiante, que quiere explorar su objeto de estudio con las diferentes técnicas posibles. De esta forma, a quien esté interesado se le facilitará acercarse al mundo del color, a través de la historia natural
de la civilización. Mediante una breve revisión, se ofrecerán las teorías que explican
cómo se ve el color y cómo lo percibimos. Asimismo, poco a poco se adentrará en
las vicisitudes técnicas necesarias para conocer el nombre origen, la química de
los compuestos, fórmulas, sinónimos, además de las metodologías más empleadas
para su aplicación en los tejidos orgánicos.
La histología, como ciencia que explora y explica la morfología microscópica, tiene
a la microtecnia como herramienta fundamental para la creación del artefacto y la
preparación permanente o “laminilla”, que permite la observación. Esta mancuerna
de ciencia y técnica ha producido algunos de los descubrimientos más importantes
de las ciencias naturales, específicamente, en biología y medicina.
La histología ha creado una forma de ver la estructura fundamental de los organismos, y los histólogos e histopatólogos han creado un lenguaje semiótico universal
que les permite la comunicación entre ellos. Este lenguaje semiótico está conformado por dos cuerpos: el lenguaje oral o escrito, aparentemente críptico, pero
puede ser desvelado mediante el otro cuerpo, el lenguaje de las imágenes, de lo
visual, de lo sensual. Estas imágenes provocan un irremediable impacto en la percepción sensorial de cualquier ser humano, y esta percepción aumentada se crea
a través de un objeto creado exprofeso, que puede ser entendido dentro de una
configuración imaginaria de realidades no visibles, pero con varias posibilidades y
con una diversidad de resultados legítimos posibles.
Este texto no es una guía final de los colorantes o de lo que se conoce sobre ellos,
solo pretende ser un instrumento, que puede ser mejorado a lo largo del tiempo, y
con su uso en los laboratorios que han adquirido, como feliz compromiso, revelar
la maravillosa variedad que se esconde en cualquier ser vivo, así como en sus
componentes más finos.
... There is a constant pursuit in modern biomedical research to observe and record processes on the cellular and intercellular level while minimizing any external influences on the object of interest [1,2]. Especially in the field of histology, advances are facilitated by enhancements and developments in microscopy methods [3,4]. Conventional methods of cultivating cell cultures have limited growth to virtually two spatial dimensions, effectively blocking cells from developing and interacting as they would in tissue. ...
Advances in imaging of microscopic structures are supported and complemented by adaptive visualization tools. These tools enable researchers to precisely capture and analyze complex three-dimensional structures of different kinds such as crystals, microchannels and electronic or biological material. In this contribution, we focus on 3D cell cultures. The new possibilities can play a particularly important role in biomedical research, especially here in the study of 3D cell cultures such as spheroids in the field of histology. By applying advanced imaging techniques, detailed information about the spatial arrangement and interactions between cells can be obtained. These insights help to gain a better understanding of cellular organization and function and have potential implications for the development of new therapies and drugs. In this context, this study presents a multi-modal light sheet microscope designed for the detection of elastic and inelastic light scattering, particularly Rayleigh scattering as well as the Stokes Raman effect and fluorescence for imaging purposes. By combining multiple modalities and stitching their individual results, three-dimensional objects are created combining complementary information for greater insight into spatial and molecular information. The individual components of the microscope are specifically selected to this end. Both Rayleigh and Stokes Raman scattering are inherent molecule properties and accordingly facilitate marker-free imaging. Consequently, altering influences on the sample by external factors are minimized. Furthermore, this article will give an outlook on possible future applications of the prototype microscope.
... There is a constant pursuit in modern biomedical research to observe and record processes on the cellular and intercellular level while minimizing any external influences on the object of interest [1,2]. Especially in the field of histology, advances are facilitated by enhancements and developments in microscopy methods [3,4]. Conventional methods of cultivating cell cultures have limited growth to virtually two spatial dimensions, effectively blocking cells from developing and interacting as they would in tissue. ...
Advances in imaging of microscopic structures are supported and complemented by adaptive visualization tools. These tools enable researchers to precisely capture and analyze complex three-dimensional structures of different kinds such as crystals, microchannels and electronic or biological material. In this contribution, we focus on 3D cell cultures. The new possibilities can play a particularly important role in biomedical research, especially here in the study of 3D cell cultures such as spheroids in the field of histology. By applying advanced imaging techniques, detailed information about the spatial arrangement and interactions between cells can be obtained. These insights help to gain a better understanding of cellular organization and function and have potential implications for the development of new therapies and drugs. In this context, this study presents a multi-modal light sheet microscope designed for the detection of elastic and inelastic light scattering, particularly Rayleigh scattering as well as the Stokes Raman effect and fluorescence for imaging purposes. By combining multiple modalities and stitching their individual results, three-dimensional objects are created combining complementary information for greater insight into spatial and molecular information. The individual components of the microscope are specifically selected to this end. Both Rayleigh and Stokes Raman scattering are inherent molecule properties and accordingly facilitate marker-free imaging. Consequently, altering influences on the sample by external factors are minimized. Furthermore, this article will give an outlook on possible future applications of the prototype microscope.
... 3.6.9. Histopathological Examination For the detection of pathological changes in the brains of mice infected with the Me49 Toxoplasma strain, a part of the brain tissues from all studied groups was fixed in 10 % formalin using hematoxylin and eosin stain (H&E) [45]. ...
The hydrazones 3a–c, were synthesized from the reaction of indole-3-carbaldehyde and nicotinic acid hydrazide, isonicotinic acid hydrazide, and benzoic acid hydrazide, respectively. Their structures were confirmed using FTIR, ¹HNMR, and ¹³CNMR spectroscopic techniques. Exclusively, hydrazones 3b and 3c were confirmed using single crystal X-ray crystallography to exist in the Eanti form. With the aid of DFT calculations, the most stable configuration of the hydrazones 3a–c in gas phase and in nonpolar solvents (CCl4 and cyclohexane) is the ESyn form. Interestingly, the DFT calculations indicated the extrastability of the EAnti in polar aprotic (DMSO) and polar protic (ethanol) solvents. Hirshfeld topology analysis revealed the importance of the N…H, O…H, H…C, and π…π intermolecular interactions in the molecular packing of the studied systems. Distribution of the atomic charges for the hydrazones 3a–c was presented. The hydrazones 3a–c showed a polar character where 3b has the highest polarity of 5.7234 Debye compared to the 3a (4.0533 Debye) and 3c (5.3099 Debye). Regarding the anti-toxoplasma activity, all the detected results verified that 3c had a powerful activity against chronic toxoplasma infection. Compound 3c showed a considerable significant reduction percent of cyst burden in brain homogenates of toxoplasma infected mice representing 49%.
... Various stains, such as blue-eosin and Masson's trichomes, have become popular in modern histology. Trichrome stain shows how complicated the method of coloring is in looking for efficient and consistent coloring that will show network refined and differentiated [89]. ...
Diabetic nephropathy (DN) is the most common complication of diabetes mellitus (DM). It is characterized by high blood glucose levels or hyperglycemia and is accompanied by changes in lipid, carbohydrate, and protein metabolism which can lead to an increased risk of complications due to vascular disease. DN is probably the most insidious among these complications, causing substantial morbidity and mortality. In this article, we will review the literature on animal models of diabetes. We will discuss several species as animal models for Type 1 and 2 diabetes, including zebrafish, rabbits, mice, rats, and rat models. This article also provides various methods used in research with model animals and presents the required result for studying diabetic DN.
... Biological samples have limited intrinsic contrast in the aforementioned imaging modalities. Therefore, an extensive set of extrinsic contrast agents 4 and genetically encoded tools such as Green Fluorescent Protein (GFP) 2,5 that produce intrinsic contrast are now available. ...
A cloneable NanoParticle (cNP) is an inorganic nanoparticle that is synthesized by a protein. The protein determines the elemental composition, size, morphology, and other properties of the nanoparticle. Here, we describe the use of a cloneable Selenium NanoParticle (cSeNP) as a cloneable imaging contrast agent in electron microscopy, fluorescence microscopy, and X-ray computed tomography. Combined, these three imaging modalities produce images of biological length scales spanning meters to angstroms but are difficult to correlate. The cSeNP is comprised of an NADPH dependent enzyme that reduces selenodiglutathione to zerovalent Se precipitates. SeNP binding peptides fused to the enzyme ligate the Se precipitates, retain them at the enzyme, and regulate SeNP size to ~5 nm inside cells. The cSeNP-protein can be genetically fused to any protein-of-interest, creating a chimeric protein-of-interest-cSeNP. The cSeNPs endow the protein-of-interest with distinguishable contrast in X-ray and EM images, due to the contrast of SeNPs in these imaging modalities relative to biological background. The cSeNPs can react spontaneously with transition metals such as Znn+ or Cun+, forming fluorescent metal-selenides, imageable in florescence microscopy. The cSeNP, therefore, represents a cloneable imaging contrast agent that facilitates location and correlation of proteins-of-interest across all biological length scales.
... Biopsy is a diagnostic technique involving the collection of the tissue samples and investigation using a microscope. The examination of tissue related to disease is called histopathology [6]. The diagnosis of breast cancer in terms of grading and staging is carried out by a pathologist through visual inspection of histopathological samples using a microscope [7]. ...
This study assesses the impact of using an Adaptive Mean Filter (AMF) as a preprocessing stage for classification of breast tumor histopathological images at various magnifications. The histopathological image was converted from red-green-blue (RGB) into grayscale before AMF is applied. In this study, AMF was performed with kernel sizes of 3 × 3 and 5 × 5 pixels. The datasets were extracted using transfer learning VGG16 before being classified using Bagging classifier. To obtain unbiased performance of the model, stratified K fold cross-validation with K = 10 was used. The dataset was divided into K-equal-sized folds. For each fold, the model was trained on the remaining K-1 folds then evaluated on the held-out fold. This process was repeated K times, with each fold used once as the validation set. The accuracy of the model was then averaged over the K folds to estimate its generalization performance. The AMF with a kernel size of 3 × 3 pixels improves the multi-class classification accuracy for magnifications of 40× and 200×, resulting in accuracy increases of 0.20% and 0.89%, respectively. However, at a magnification of 100×, the model's performance decreases. While the use of AMF with a kernel size of 3 × 3 pixels did not raise the accuracy at magnification 400×, it resulted in a lower standard deviation by 0.24%. In binary-class classification, the use of the AMF with a kernel size of 3 × 3 pixels improves accuracy by 1.10% for magnification 40× and by 0.85% for magnification 200×. However, when implemented at magnifications of 100× and 400×, the AMF filter results in decreased performance. In conclusion, the use of the AMF with a kernel size of 3 × 3 pixels as a preprocessing stage for the histopathological image classification of breast tumor has shown to have a positive impact on the accuracy of multi-class and binary-class classifications for magnifications of 40× and 200×, but not for magnifications of 100× and 400×. The results also indicate that the use of Publication Issue 275 AMF filter can reduce the standard deviation compared to without AMF for some magnifications. However, caution should be considered when applying the AMF filter, as it can decrease the model performance in some cases.
... Man has always sought for ways to improve his health quality, eradicate, and prevent disease conditions that always attack him, and for many years, natural products have been recognised as one of the major sources of medicine across the world for management and prevention of diseases. [1][2][3][4] Also, over the years, the use of plant derived drugs has proven to be a good choice of drug for all kinds of diseases as it has better cultural adaptability with the human body and pose lesser side effects, 5 as compared to conventional drugs that have shown incidences of relapse, side effects and drug-body interaction, 5,6 Newman and Cragg 3 reported that prior to 2007, about 50% of all standard drugs registered over a 25-year timeline were sourced from natural products and their synthetic derivatives. Histology, originally derived from the Greek root words; histos (tissues) and logos (study) 7 is a science that deals with the study of body organs and tissues at the microscopic level. ...
This study was aimed to investigate the histological effect and antimicrobial activity of a compound [3,5,6,7-tetrahydroxy-19-vouacanoic acid;(3β, 5α, 6β, 7β)-form,6,7-dibenzoyl] previously isolated from Caesalpinia pulcherrima stem bark prior to this study. Different doses (2, 4 and 8 mg/kg) of the compound was administered to Wistar rats (both sexes) for a 28-day period. After the 28-day administration, vital organs (liver, heart, kidney, lung, uterus and testes) were harvested from the animals to ascertain their organ weight variations and histology. Microbial activity of the compound was investigated (using zone of inhibition test and minimum inhibitory concentration) for the microorganisms (Bacillus subtilis, Klebsiella pneumoniae, P. aeruginosa, Staphylococcus aureus, Candida. albican and Trychophyton rubrum). The observed weight changes in organs for all treated groups in relative to control was not significant at p < 0.05 and no observed toxic abnormalities for all organs in treated groups upon their gross examination in relative to control. Highest zone inhibition was observed at 100 mg/kg for all tested microorganisms (B. subtilis, K. pneumonia, P. aeruginosa, S. aureus, C. albican and T. rubrum). with their corresponding MICs 11, 13, 16, 33, 30 and 31 mg/ml respectively. This study suggests that the compound could be a potential candidate in view of developing new antimicrobial agent and potential novel drug as a vasodilator.
... On day 70 of the experiment, a portion of the brain tissues from all studied groups were fixed in 10 % formalin solution and assessed for any pathological findings using hematoxylin and eosin (H&E) staining. Grocott-Gomori methenamine silver stain (GMS) was used to stain the cyst wall and bradyzoites [34,35]. ...
The tight relationship between immunity and retinoid levels provides evidence on the critical role of retinoic acid (RA) in regulating immune activity, especially the mucosal one. Mucosal immune response is the key for determination of the outcome of infection, particularly against intracellular mucosal pathogens such as Toxoplasma gondii, where it plays a crucial role as a sentinel against parasite invasion. Herein, the immunomodulatory adjuvant role of RA was evaluated for prophylactic vaccination against chronic Toxoplasma infection. A quantity of 15 µg of RA pre-encapsulated with lipid-based nanoparticles (SLNs) was intranasally used in three doses, two weeks apart, as an adjuvant to the Toxoplasma lysate antigen (TLA). Afterward, mice were infected with 20 cysts of T. gondii (ME49 strain) and were sacrificed at the 4th week post-infection. Parasitological, immunological, biochemical, and histopathological studies were applied as vaccine efficacy measures. The protective role of the tested vaccine was noted using the statistically marked reduction in brain cyst count, accompanied by remarkable levels of protective IFN-γ and antibodies, with amelioration of infection-induced oxidative stress and brain pathology. Ultimately, this experiment outlined the prospective role of a novel, natural, nano-encapsulated and mucosal vaccine adjuvant RA-SLNs as a propitious candidate against chronic toxoplasmosis.
... There was no clear difference in the number of degenerated cells between the three doses, but there was an impression at medium and high doses on the histopathological picture. It is difficult to distinguish the proportion of pancreatic cell damage in the assessment with hematoxylin and eosin staining accurately between each treatment level of bitter melon fruit because the assessment with hematoxylin-eosin staining is less sensitive in detecting endocrine cell degeneration, and is not a specific stain [21]. ...
This study aims to determine the class of chemical compounds, phytochemical levels, antihyperglycemic effects, and histopathology of the pancreas of rats treated with bitter gourd (Momordica charantia L) for the treatment of diabetes mellitus. Bitter gourd is made by adding water, then mashed and squeezed. The ingredients were tested for their phytochemical content and antidiabetic power. 15 male rats were divided into 5 groups. Group 1 as a negative control, namely normal rats without treatment, group 2 as a positive control given alloxan, group 3, namely rats given alloxan and bitter melon fruit 0.9 mL, group 4, namely rats given alloxan and bitter melon ingredients 1.8 mL, and group 5, namely rats, were given alloxan and a mixture of bitter melon 3.6 mL. The phytochemical test results of bitter melon fruit contain carbohydrates, alkaloids, saponins, and terpenoids. The total alkaloid content of the bitter melon fruit is 0.232%, the saponin content of the bitter melon fruit is 0.6375%, and the terpenoid content of the bitter melon fruit is 0.5317%. The administration of bitter melon fruit to diabetic rats can reduce blood sugar levels of rats and the histopathological observations of the pancreas of rats show a repair effect on pancreatic damage after administration of bitter melon herb.
... Dalam kasus diagnosis medis tomor, pewarnaan adalah proses medis umum yang dilakukan. Pewarnaan tersebut dapat menjelaskan batasan posterior dan anterior jaringan sampel untuk menemukan sel yang sakit atau tumor atau sel patologis lainnya (Musumeci, 2014). Dalam studi biologi, pewarnaan digunakan untuk menandai sel dan menandai asam nukleat, protein, atau elektroforesis gel untuk membantu pemeriksaan mikroskopis (Yadav et al., 2019). ...
Xylol adalah agen deparafinisasi yang umum digunakan ditiap tahapan deparafinisasi pada pewarnaan histologi jaringan. Namun karena sifat toksisitas tinggi dan harga yang mahal, maka diupayakan mencari alternatif yang lebih aman dan murah. Ez Prep concentrate adalah salah satu agen deparafinisasi yang umum digunakan pada alat Ventana Medical Systems, Inc (Ventana). Produk ini dirancang untuk digunakan sebagai pewarna slide otomatis BenchMark Series. Selain harga nya lebih murah dibanding xylol, toksisitasnya juga rendah. Perlu dibuktikan efektifitas reagen EZ Prep concentrate agar dapat digunakan sebagai alternatif pengganti xylol. Penelitian ini bersifat eksperimen yang dilaksanakan pada bulan September – November 2022 di Laboratorium Patologi Anatomi Siloam Hospital Lippo Village. Sampel yang digunakan jaringan miom dan payudara. Hasil penelitian menunjukkan semua sampel yang dideparfinasi dengan EZ prep concentrate memberikan kualitas hasil mikroskopis yang baik (skor 3). Maka, EZ Prep concentrate dapat dipertimbangakan penggunaannya sebagai pengganti xylol.
... Part of the brain tissues harvested at day 70 from all studied groups was fixed in 10 % formalin for histopathological examination using hematoxylin and eosin (H&E). Pathological changes in the brains of mice challenged with the Me49 Toxoplasma strain were recorded [35]. ...
Melatonin (MLT) is now emerging as one of the universally accepted immunostimulators with broad applications in medicine. It is a biological manipulator of the immune system, including mucosal ones. MLT was encapsulated in solid lipid nanoparticles (SLNs), then 100 mg/kg/dose of MLT-SLNs was used as an adjuvant of Toxoplasma lysate antigen (TLA). Experimental mice were intra-nasally inoculated with three doses of different regimens every two weeks, then challenged with 20 cysts of T. gondii Me49 strain, where they were sacrificed four weeks post-infection. Protective vaccine efficacy was evident via the significant brain cyst count reduction of 58.6%, together with remarkably high levels of humoral systemic and mucosal anti-Toxoplasma antibodies (Ig G, Ig A), supported by a reduced tachyzoites invasion of Vero cells in vitro upon incubation with sera obtained from these vaccinated mice. A cellular immune response was evident through the induction of significant levels of interferon-gamma (IFN γ), associated with morphological deteriorations of cysts harvested from the brains of vaccinated mice. Furthermore, the amelioration of infection-induced oxidative stress (OS) and histopathological changes were evident in mice immunized with TLA/MLT-SLNs. In conclusion, the present study highlighted the promising role of intranasal MLT-SLNs as a novel mucosal adjuvant candidate against chronic toxoplasmosis.
... The cells were then collected and plated in six-well plates at a density of 5 × 10 2 cells per well. After 10 days, cells were washed with phosphate-buffered saline, followed by fixation with methanol/acetic acid (3:1; v:v), and finally stained with Giemsa [18]. The number of colonies were counted under a microscope [19]. ...
Background
Poly(A)-binding protein interacting protein 1 (PAIP1) is a translational initiation regulatory factor that has been reported as oncogene in multiple malignant diseases. However, its role in hepatocellular carcinoma (HCC) and the potential mechanisms have not been explored.
Methods
PAIP1 expression level in HCC cell lines were detected by real-time quantitative PCR and western blotting. The proliferation and colony formation of HCC cell lines were detected by MTT and colony formation assay. The apoptosis and cell cycle were detected by flow cytometry. The volume and growth rate of the xenograft tumors were observed. The potential mechanism of PAIP1 was analyzed by miRNA Microarray Analysis and TargetScan analysis.
Results
PAIP1 is significantly upregulated in HCC cell lines. PAIP1 knockdown dramatically inhibits cell proliferation and colony formation, induces apoptosis and alters the cell cycle distribution by increasing the G2/M cell percentage. Moreover, PAIP1 knockdown significantly reduces tumorigenesis in a murine transplantation model. Bioinformatics and immunoblotting analysis reveal that PAIP1 knockdown dysregulates cyclin D pathway-related proteins.
Conclusion
PAIP1 plays an oncogenic role in hepatocellular carcinoma.
... 20,28,29 Moreover, it has been demonstrated that a dermatopathologist's tendency towards rendering a malignant as opposed to non-malignant (i.e., atypical) diagnosis is subject to diagnostic drift favoring the former over time. 30 Thus, although the human microscopic evaluation of hematoxylin and eosin stained tissue sections has been the cornerstone of histological cancer diagnoses for the past century, 25,31,32 the need for more objective and standardized methods of histochemical detection and classification that meet the current demand is of paramount clinical concern. 23,31 Digital pathology-the practice of using high-resolution digital imaging to manage and analyze data from digitized specimen slides-is rapidly becoming the standard of care in the field of pathology. ...
Background
Skin cancers are the most common malignancies diagnosed worldwide. While the early detection and treatment of pre-cancerous and cancerous skin lesions can dramatically improve outcomes, factors such as a global shortage of pathologists, increased workloads, and high rates of diagnostic discordance underscore the need for techniques that improve pathology workflows. Although AI models are now being used to classify lesions from whole slide images (WSIs), diagnostic performance rarely surpasses that of expert pathologists.
Objectives
The objective of the present study was to create an AI model to detect and classify skin lesions with a higher degree of sensitivity than previously demonstrated, with potential to match and eventually surpass expert pathologists to improve clinical workflows.
Methods
We combined supervised learning (SL) with semi-supervised learning (SSL) to produce an end-to-end multi-level skin detection system that not only detects 5 main types of skin lesions with high sensitivity and specificity, but also subtypes, localizes, and provides margin status to evaluate the proximity of the lesion to non-epidermal margins. The Supervised Training Subset consisted of 2188 random WSIs collected by the PathologyWatch (PW) laboratory between 2013 and 2018, while the Weakly Supervised Subset consisted of 5161 WSIs from daily case specimens. The Validation Set consisted of 250 curated daily case WSIs obtained from the PW tissue archives and included 50 “mimickers”. The Testing Set (3821 WSIs) was composed of non-curated daily case specimens collected from July 20, 2021 to August 20, 2021 from PW laboratories.
Results
The performance characteristics of our AI model (i.e., Mihm) were assessed retrospectively by running the Testing Set through the Mihm Evaluation Pipeline. Our results show that the sensitivity of Mihm in classifying melanocytic lesions, basal cell carcinoma, and atypical squamous lesions, verruca vulgaris, and seborrheic keratosis was 98.91% (95% CI: 98.27%, 99.55%), 97.24% (95% CI: 96.15%, 98.33%), 95.26% (95% CI: 93.79%, 96.73%), 93.50% (95% CI: 89.14%, 97.86%), and 86.91% (95% CI: 82.13%, 91.69%), respectively. Additionally, our multi-level (i.e., patch-level, ROI-level, and WSI-level) detection algorithm includes a qualitative feature that subtypes lesions, an AI overlay in the front-end digital display that localizes diagnostic ROIs, and reports on margin status by detecting overlap between lesions and non-epidermal tissue margins.
Conclusions
Our AI model, developed in collaboration with dermatopathologists, detects 5 skin lesion types with higher sensitivity than previously published AI models, and provides end users with information such as subtyping, localization, and margin status in a front-end digital display. Our end-to-end system has the potential to improve pathology workflows by increasing diagnostic accuracy, expediting the course of patient care, and ultimately improving patient outcomes.
... Over the past century, histological staining has been established as a principal tool for tissue examination in disease diagnostics and life-science research 1,2 . By labeling different biological elements with specific markers based on their biochemical properties, histological staining enables the visualization of tissue and cellular structures and allows the assessment of pathophysiology and disease development when the stained samples are observed under a light microscope [3][4][5] . ...
Histological staining is the gold standard for tissue examination in clinical pathology and life-science research, which visualizes the tissue and cellular structures using chromatic dyes or fluorescence labels to aid the microscopic assessment of tissue. However, the current histological staining workflow requires tedious sample preparation steps, specialized laboratory infrastructure, and trained histotechnologists, making it expensive, time-consuming, and not accessible in resource-limited settings. Deep learning techniques created new opportunities to revolutionize staining methods by digitally generating histological stains using trained neural networks, providing rapid, cost-effective, and accurate alternatives to standard chemical staining methods. These techniques, broadly referred to as virtual staining, were extensively explored by multiple research groups and demonstrated to be successful in generating various types of histological stains from label-free microscopic images of unstained samples; similar approaches were also used for transforming images of an already stained tissue sample into another type of stain, performing virtual stain-to-stain transformations. In this Review, we provide a comprehensive overview of the recent research advances in deep learning-enabled virtual histological staining techniques. The basic concepts and the typical workflow of virtual staining are introduced, followed by a discussion of representative works and their technical innovations. We also share our perspectives on the future of this emerging field, aiming to inspire readers from diverse scientific fields to further expand the scope of deep learning-enabled virtual histological staining techniques and their applications.
... Herbal medicines related to hepatotoxicity is the second most common cause of drug-induced liver injury nowadays. Histological examination is the benchmark for assessing treatment associated to pathological changes in tissues and organs (Haouas et al., 2014;Musumeci, 2014;Shubin et al., 2016). Histology of kidneys in both studies showed no significant change of lesion scores. ...
... In animal studies, the histological examination is widely used to detect signs of disease not easily recognised by gross examination and is a vital means of supervising general fish health (Gallhoefer et al., 2013;Musumeci, 2014). ...
We designed this experiment to examine the impact of high levels of dietary Cr(III) on the histological structure of the gut, liver, kidney, and spleen of the juvenile common carp (Cyprinus carpio L.; average weight 15 ± 0.69 g). Fish were fed with different levels of dietary Cr as chromium chloride CrCl3.6H2O (0.0, 4.0, 6.0, and 8.0) mg kg-1 diet for 10 weeks at 25 ± 1.5ºC. At the end of the experiment, no mortalities were recorded. High levels of dietary Cr impaired the normal histological structure of the tested organs. Different Cr treatments showed signs of toxicity in the liver and gut that involved the appearance of inflammatory foci cells, pyknotic nuclei, foci of melanomacrophages, vacuole formation, degeneration of hepatocytes and erythrocytes aggregation in the liver and lifting of the lining epithelium, vacuolation, and surface erosion in the intestine. Cr treatments showed remarkable changes in the kidney that included renal tubular separation, cytoplasmic vacuolation, aggregation of red blood cells, degeneration of renal tubules, necrosis of hematopoietic tissue, and oedema. Injuries and damage were observed in the spleens of fish fed with Cr-fortified diets, including necrosis, depletion of lymphoid tissues, and vacuoles formation. In conclusion, ingestion of high levels of dietary Cr(III) generated adverse health effects, and the application of Cr(III) in fish diets should be cautious.
... Spleen sections were assessed for architecture, changes in red and white pulp and presence/absence of tachyzoites. Brain sections were also examined in different groups [64,65]. ...
Toxoplasma gondii (T. gondii) is a highly prevalent parasite that has no gold standard treatment due to the poor action or the numerous side effects. Focused sulfonamide-1,2,3-triazole hybrids 3a–c were wisely designed and synthesized via copper catalyzed 1,3-dipolar cycloaddition approach between prop-2-yn-1-alcohol 1 and sulfa drug azides 2a–c. The newly synthesized click products were fully characterized using different spectroscopic experiments and were loaded onto chitosan nanoparticles to form novel nanoformulations for further anti-Toxoplasma investigation. The current study proved the anti-Toxoplasma effectiveness of all examined compounds in experimentally infected mice. Relative to sulfadiazine, the synthesized sulfonamide-1,2,3-triazole (3c) nanoformulae demonstrated the most promising result for toxoplasmosis treatment as it resulted in 100% survival, 100% parasite reduction along with the remarkable histopathological improvement in all the studied organs.
The gold standard for studying biological soft tissues at the microscale (i.e., histology) is tissue sectioning with subsequent colorimetric or fluorescent staining and visual inspection under the microscope. When tissue integrity must be maintained for 3D histological assessment, contrast-enhanced microfocus X-ray computed tomography (CECT) is a promising solution, but there is still a lack of staining protocol optimization of contrast-enhancing staining agents (CESAs). Therefore, in this study, mouse auricles were incubated with Hafnium-substituted Wells-Dawson polyoxometalate, cationic iodinated contrast agent, or Lugol's iodine and were imaged with high-resolution CECT. Alignment with corresponding H&E-stained sections enabled the identification and segmentation of different tissue types. Contrast differences between tissue types were increased by washing the samples after staining or by combining CESAs. Finally, we proved that the latter could be used to quantitatively assess the 3D thickness distribution of the epidermis in the ears of a mouse model of psoriasis-like dermatitis. In conclusion, CECT and bright-field microscopy are complementary and not mutually exclusive techniques for the histological assessment of biological tissues. While bright-field microscopy gives detailed information about the cellular composition of tissues, CECT provides a better insight into the spatial interrelationship of tissues and is a powerful tool for performing 3D structural quantification.
Many developments occurred in the later years of the twentieth century and the early years of the twenty-first century that have contributed to improved diagnosis and treatment of patients with bone tumors and that have advanced the specialties of bone pathology and skeletal radiology. These include immunohistochemistry, genetic analysis, liquid biopsy, CT, MRI, PET scanning, and artificial intelligence, among others.
In pathological diagnostics, histological images highlight the oncological features of excised specimens, but they require laborious and costly staining procedures. Despite recent innovations in label-free microscopy that simplify complex staining procedures, technical limitations and inadequate histological visualization are still problems in clinical settings. Here, we demonstrate an interconnected deep learning (DL)-based framework for performing automated virtual staining, segmentation, and classification in label-free photoacoustic histology (PAH) of human specimens. The framework comprises three components: (1) an explainable contrastive unpaired translation (E-CUT) method for virtual H&E (VHE) staining, (2) an U-net architecture for feature segmentation, and (3) a DL-based stepwise feature fusion method (StepFF) for classification. The framework demonstrates promising performance at each step of its application to human liver cancers. In virtual staining, the E-CUT preserves the morphological aspects of the cell nucleus and cytoplasm, making VHE images highly similar to real H&E ones. In segmentation, various features (e.g., the cell area, number of cells, and the distance between cell nuclei) have been successfully segmented in VHE images. Finally, by using deep feature vectors from PAH, VHE, and segmented images, StepFF has achieved a 98.00% classification accuracy, compared to the 94.80% accuracy of conventional PAH classification. In particular, StepFF’s classification reached a sensitivity of 100% based on the evaluation of three pathologists, demonstrating its applicability in real clinical settings. This series of DL methods for label-free PAH has great potential as a practical clinical strategy for digital pathology.
Background: Histopathology and physiology play crucial roles in the diagnosis of diseases. Histopathology involves the microscopic examination of tissues, while physiology focuses on the activities of organs and systems. This paper examines the interaction between histological techniques and illness diagnosis, emphasizing the progress made in these techniques and their crucial importance in medical diagnostics. Methods: This comprehensive review analyzes the progression of histopathology from basic tissue observations to more sophisticated methods such as immunohistochemistry and digital pathology. It highlights the role of these techniques in diagnosing a range of diseases, including cancer, infectious diseases, autoimmune disorders, and neurological conditions. Results: Histopathology offers a vital understanding of disease causes and severity, necessary for precise diagnosis and efficient therapy planning. Diagnostic precision has been improved by special staining, immunohistochemistry, and molecular pathology. The study emphasizes the significance of physiological measurements in comprehending functional irregularities and the impact of diseases, aiding in the early detection and categorization of risks. Conclusion: The combination of histology and physiology provides a comprehensive approach to diagnosing and treating diseases. The collaboration across these disciplines is crucial for advancing targeted medicines and enhancing patient outcomes, highlighting the ongoing development and significance of these fields in contemporary medicine. Article Info.
The routine staining which is also called as H & E staining (Hematoxylin and eosin) is the most widely usedstain in the laboratories or histopathology laboratory. In this procedure the nuclear protein stains purpleand the cytoplasm and any other substances stains as red or orange. It plays a very crucial role in medicaldiagnosis or research where the specimen or tissue is been sent for processing. The staining procedure isperformed by trained professionals and after which the role of highly trained pathologists and researcherscome, which under the microscope view the specific structures, cells and even microorganisms to providea diagnosis for the specific disease. The special stains are those which come under the special stainingtechnique or alternative method for the H & E stains, where H & E staining does not provide the properidentification or appreciation of certain normal and abnormal tissue or structure which the pathologist orresearchers needs. This article throws light on the Special stains which are been used in histopathologicalprocedures in which these abnormal or normal tissues can be properly appreciated. The article also focuseson the procedure, principle and uses of these special stains in their respective field.
Histology and embryology have evolved over time and are now in their 4.0 stages. These fields have progressed from basic microscopes and staining techniques to advanced imaging and computational approaches. Histology 4.0 combines digital imaging technology, AI, and big data analytics to improve tissue analysis accuracy, reduce costs, and develop personalized treatment plans. Embryology 4.0 uses advanced imaging, molecular biology, and bioinformatics to understand developmental biology and identify therapeutic targets for regenerative medicine. Both fields have potential applications in various areas of medicine and are expected to further advance with continued development of new technologies and approaches.
Infrared (IR) spectroscopic imaging records spatially resolved molecular vibrational spectra, enabling a comprehensive measurement of the chemical makeup and heterogeneity of biological tissues. Combining this novel contrast mechanism in microscopy with the use of artificial intelligence can transform the practice of histopathology, which currently relies largely on human examination of morphologic patterns within stained tissue. First, this review summarizes IR imaging instrumentation especially suited to histopathology, analyses of its performance, and major trends. Second, an overview of data processing methods and application of machine learning is given, with an emphasis on the emerging use of deep learning. Third, a discussion on workflows in pathology is provided, with four categories proposed based on the complexity of methods and the analytical performance needed. Last, a set of guidelines, termed experimental and analytical specifications for spectroscopic imaging in histopathology, are proposed to help standardize the diversity of approaches in this emerging area.
Expected final online publication date for the Annual Review of Analytical Chemistry, Volume 16 is June 2023. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.
Histological staining is the gold standard for tissue examination in clinical pathology and life-science research, which visualizes the tissue and cellular structures using chromatic dyes or fluorescence labels to aid the microscopic assessment of tissue. However, the current histological staining workflow requires tedious sample preparation steps, specialized laboratory infrastructure, and trained histotechnologists, making it expensive, time-consuming, and not accessible in resource-limited settings. Deep learning techniques created new opportunities to revolutionize staining methods by digitally generating histological stains using trained neural networks, providing rapid, cost-effective, and accurate alternatives to standard chemical staining methods. These techniques, broadly referred to as virtual staining, were extensively explored by multiple research groups and demonstrated to be successful in generating various types of histological stains from label-free microscopic images of unstained samples; similar approaches were also used for transforming images of an already stained tissue sample into another type of stain, performing virtual stain-to-stain transformations. In this Review, we provide a comprehensive overview of the recent research advances in deep learning-enabled virtual histological staining techniques. The basic concepts and the typical workflow of virtual staining are introduced, followed by a discussion of representative works and their technical innovations. We also share our perspectives on the future of this emerging field, aiming to inspire readers from diverse scientific fields to further expand the scope of deep learning-enabled virtual histological staining techniques and their applications. Deep Learning enables virtual histological staining of biological samples.
Special stains are used to visualize various tissue elements and entities.They provide valuable information in the evaluation of various disease conditions.The principle on which they work is the interaction of intracellular and extracellular chemical reactions between the tissue components and the dyes.They allow the target substance to be identified on the basis of their chemical and biological character and thus help in diagnostic research.
Many of the time-honored methods for histopathological analysis of tissue sections have been superseded by new histological techniques, immunohistochemistry, or other diagnostic tests. Some of the dyes that were used for generations are no longer available, and chemicals used in some older methods are now considered hazardous. Additionally, the histotechnologists and pathologists familiar with these techniques are now retiring and leaving the field, resulting in a loss of experience. To provide a sense of the rich legacy of histopathological analysis, this article reviews the development of special stains in histopathology, describes staining techniques, and discusses the wide range of applications. Numerous staining techniques mentioned in this article were originally published many years ago, some in obscure journals in foreign countries, and are not readily available to practicing histotechnologists. To encourage further study, references in English are given, many to popular histotechnology texts found in most histopathology laboratories or biomedical libraries. (The J Histotechnol 32(1):9–19, 2009)Submitted June 5, 2008; accepted with revisions September 1, 2008
The hospital-based histology laboratory experienced several changes in the 1960s and 1970s when cryostats, enclosed tissue processors, plastic cassettes, and disposable knives were introduced. Other than the emergence of immunohistochemistry in the 1980s, few major changes have occurred. But now, driven by costs, higher expectations, and a desire for faster turnaround times, these laboratories are on the brink of changes that will involve additional new duties for the pathologist and the histotechnologist.
New techniques, some already in use in some hospitals, will involve flow cytometry, fluorescence in situ hybridization (FISH), DNA and genetics studies, proteomics, telepathology, and digital imaging. New antibodies in immunohistochemistry and new biomarkers will assist in diagnoses. Accrediting agencies will raise their standards to meet the new challenges. Although no one really knows what will happen in the future, this article reviews the new changes that will likely occur in hospital histology, based on present predictions and technology.
The specialty of histopathology technique dates back to 1838, when Johannes Miiller published his book, On the Nature and Structure Characteristics of Cancer, the first book on histopathology. The first compound microscope had been constructed earlier in 1591 but suffered from severe optical problems. In 1673 Anton van Leeuwenhoek started the development of simple microscopes with single lenses but that gave improved magnification and resolution. The first microtome suitable for sectioning animal tissues was constructed in 1848, with the popular Cambridge Rocker (1885), Minot (1886), and sledge microtomes (1910) manufactured later. Paraffin wax for infiltration and support during sectioning was introduced during the mid1800s. Different laboratory chemicals were investigated for use as fixatives. Formalin, widely used today, was first used in 1893.Automated tissue processors replaced hand processing starting in 1945, and cryostats were first manufactured in 1951. Enzyme histochemistry, electron microscopy, and polarizing microscopy all have become diagnostic tools during the last 50 years. The widespread use of immunohistochemistry began in the 1980s has revolutionized cancer diagnosis and is still under development. This article briefly reviews the development of histopathology techniques in the United States and United Kingdom from historical times to present. (The J Histotechnol 29:99, 2006)Submitted December 21, 2005; accepted with revisions March 28, 2006
Histochemistry has a remarkable long-term impact on cell and tissue biology, embryology, and pathology and remains at the forefront of research in these disciplines. Histochemical techniques, and particularly immunohistochemical and in situ hybridization techniques, are now more widely used than ever and, in conjunction with new developments in microscopical imaging and analysis, will continue to have an important position in the life sciences and medicine. However, histochemistry is often mistakenly perceived as an archaic discipline, and its contributions to cell and molecular biology are not always given the credit it deserves.
Eponyms in histology and histochemistry: do they still serve a purpose, or should they be abandoned in favor of standard noneponymous terminology? Acta Histochem | Article | PubMed Citation: Musumeci G. Past, present and future: overview on histology and histopathology
- R Coleman
Coleman R. Eponyms in histology and histochemistry: do they still serve
a purpose, or should they be abandoned in favor of standard noneponymous terminology? Acta Histochem. 2006; 108:241-2. | Article |
PubMed
Citation:
Musumeci G. Past, present and future: overview on
histology and histopathology. J Histol Histopathol.
2014; 1:5. http://dx.doi.org/10.7243/2055-091X-1-5