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Global Journal of Health Science Jun 2015The history of histology indicates that there have been significant changes in the techniques used for histological staining through chemical, molecular biology assays... (Review)
Review
The history of histology indicates that there have been significant changes in the techniques used for histological staining through chemical, molecular biology assays and immunological techniques, collectively referred to as histochemistry. Early histologists used the readily available chemicals to prepare tissues for microscopic studies; these laboratory chemicals were potassium dichromate, alcohol and the mercuric chloride to harden cellular tissues. Staining techniques used were carmine, silver nitrate, Giemsa, Trichrome Stains, Gram Stain and Hematoxylin among others. The purpose of this research was to assess past and current literature reviews, as well as case studies, with the aim of informing ways in which histological stains have been improved in the modern age. Results from the literature review has indicated that there has been an improvement in histopathology and histotechnology in stains used. There has been a rising need for efficient, accurate and less complex staining procedures. Many stain procedures are still in use today, and many others have been replaced with new immunostaining, molecular, non-culture and other advanced staining techniques. Some staining methods have been abandoned because the chemicals required have been medically proven to be toxic. The case studies indicated that in modern histology a combination of different stain techniques are used to enhance the effectiveness of the staining process. Currently, improved histological stains, have been modified and combined with other stains to improve their effectiveness.
Topics: Coloring Agents; Histological Techniques; Humans; Staining and Labeling
PubMed: 26493433
DOI: 10.5539/gjhs.v8n3p72 -
Nature Communications Aug 2021Pathology is practiced by visual inspection of histochemically stained tissue slides. While the hematoxylin and eosin (H&E) stain is most commonly used, special stains...
Pathology is practiced by visual inspection of histochemically stained tissue slides. While the hematoxylin and eosin (H&E) stain is most commonly used, special stains can provide additional contrast to different tissue components. Here, we demonstrate the utility of supervised learning-based computational stain transformation from H&E to special stains (Masson's Trichrome, periodic acid-Schiff and Jones silver stain) using kidney needle core biopsy tissue sections. Based on the evaluation by three renal pathologists, followed by adjudication by a fourth pathologist, we show that the generation of virtual special stains from existing H&E images improves the diagnosis of several non-neoplastic kidney diseases, sampled from 58 unique subjects (P = 0.0095). A second study found that the quality of the computationally generated special stains was statistically equivalent to those which were histochemically stained. This stain-to-stain transformation framework can improve preliminary diagnoses when additional special stains are needed, also providing significant savings in time and cost.
Topics: Algorithms; Biopsy, Large-Core Needle; Coloring Agents; Deep Learning; Diagnosis, Computer-Assisted; Diagnosis, Differential; Humans; Kidney; Kidney Diseases; Pathology, Clinical; Reference Standards; Reproducibility of Results; Sensitivity and Specificity; Staining and Labeling
PubMed: 34385460
DOI: 10.1038/s41467-021-25221-2 -
British Medical Journal Oct 1964
Topics: Biopsy; Child; Coloring Agents; Dermatomycoses; Fungi; Humans; India; Pathology; Staining and Labeling; Zygomycosis
PubMed: 14185623
DOI: 10.1136/bmj.2.5415.1009-b -
ELife Dec 2021Characterization and isolation of a large population of cells are indispensable procedures in biological sciences. Flow cytometry is one of the standards that offers a...
Characterization and isolation of a large population of cells are indispensable procedures in biological sciences. Flow cytometry is one of the standards that offers a method to characterize and isolate cells at high throughput. When performing flow cytometry, cells are molecularly stained with fluorescent labels to adopt biomolecular specificity which is essential for characterizing cells. However, molecular staining is costly and its chemical toxicity can cause side effects to the cells which becomes a critical issue when the cells are used downstream as medical products or for further analysis. Here, we introduce a high-throughput stain-free flow cytometry called in silico-labeled ghost cytometry which characterizes and sorts cells using machine-predicted labels. Instead of detecting molecular stains, we use machine learning to derive the molecular labels from compressive data obtained with diffractive and scattering imaging methods. By directly using the compressive 'imaging' data, our system can accurately assign the designated label to each cell in real time and perform sorting based on this judgment. With this method, we were able to distinguish different cell states, cell types derived from human induced pluripotent stem (iPS) cells, and subtypes of peripheral white blood cells using only stain-free modalities. Our method will find applications in cell manufacturing for regenerative medicine as well as in cell-based medical diagnostic assays in which fluorescence labeling of the cells is undesirable.
Topics: Coloring Agents; Computer Simulation; Flow Cytometry; Humans; Induced Pluripotent Stem Cells; Leukocytes; Machine Learning; Staining and Labeling
PubMed: 34930522
DOI: 10.7554/eLife.67660 -
Laboratory Investigation; a Journal of... May 2022Conventional histological stains, such as hematoxylin plus eosin (H&E), and immunohistochemistry (IHC) are mainstays of histology that provide complementary diagnostic...
Conventional histological stains, such as hematoxylin plus eosin (H&E), and immunohistochemistry (IHC) are mainstays of histology that provide complementary diagnostic information. H&E and IHC currently require separate slides, because the stains would otherwise obscure one another. This consumes small specimen, limiting the total amount of testing. Additionally, performing H&E and IHC on different slides does not permit comparison of staining at the single cell level, since the same cells are not present on each slide, and alignment of tissue features can be problematic due to changes in tissue landscape with sectioning. We have solved these problems by performing conventional staining and IHC on the same slide using invisible IHC chromogens, such that the chromogens are not visible when viewing the conventional stain and the conventional stain is excluded from images of the IHC. Covalently deposited chromogens provided a convenient route to invisible chromogen design and are stable to reagents used in conventional staining. A dual-camera brightfield microscope system was developed that permits simultaneous viewing of both visible conventional stains and invisible IHC chromogens. Simultaneous staining was demonstrated on several formalin-fixed paraffin-embedded tissue specimens using single and duplex IHC, with chromogens that absorb ultraviolet and near infrared light, followed by H&E staining. The concept was extended to other conventional stains, including mucicarmine special stain and Papanicoulou stain, and further extended to cytology specimens. In addition to interactive video review, images were recorded using multispectral imaging and image processing to provide flexible production of color composite images and enable quantitative analysis.
Topics: Coloring Agents; Eosine Yellowish-(YS); Hematoxylin; Immunohistochemistry; Staining and Labeling
PubMed: 34963687
DOI: 10.1038/s41374-021-00714-2 -
Acta Cytologica 2017Cytodiagnoses of specific malignancies are enabled through analyses of abnormal nuclear chromatin and cytoplasmic features in stained cells. (Review)
Review
OBJECTIVE
Cytodiagnoses of specific malignancies are enabled through analyses of abnormal nuclear chromatin and cytoplasmic features in stained cells.
AIM
The objective of this work was to explore the inception, development, and chemistry of the Pap stain method introduced in 1942 by Dr. G.N. Papanicolaou.
STUDY DESIGN
To achieve this, we carried out a review of the English literature.
RESULTS
Between 1914 and 1933, Papanicolaou first analyzed vaginal squamous cells in guinea pigs and later in human vaginal fluid samples using hematoxylin and eosin with limited color reactions, correlating the cell-type morphology with endocrinology and histology. The 5-dye Pap stain method evolved through 2 salient phases. The first, between 1933 and 1942, saw the introduction of alcohol-ether fixation and aqueous waterblue staining to enhance cellular transparency, aiding the distinction of cervical cancer cells from benign cells, with quantitative and qualitative assessment of squamous cell maturity. The second phase, between 1942 and 1960, saw the introduction and refinement of various alcoholic cytoplasmic counterstaining schemes with orange G and EA (light green, Bismarck brown, eosin) and phosphotungstic acid, allowing wider ranges of polychromasia and further enhancing cellular visualization, facilitating the distinction of cell types and improving diagnostic confidence.
CONCLUSIONS
Development of the Pap stain method followed specific historical and scientific events. The staining method evolved following incremental improvements in cellular transparency achieved through tailored cellular fixation and cytoplasmic staining using variable dye and pH combinations.
Topics: Animals; Coloring Agents; Cytoplasm; Female; Humans; Papanicolaou Test; Staining and Labeling; Uterine Cervical Neoplasms; Vaginal Smears
PubMed: 28384641
DOI: 10.1159/000457827 -
Asia-Pacific Journal of Ophthalmology... Nov 2020Chromovitrectomy, the intraocular application of dyes to assist visualization of preretinal tissues during vitreoretinal surgery, was introduced to avoid ocular... (Review)
Review
Chromovitrectomy, the intraocular application of dyes to assist visualization of preretinal tissues during vitreoretinal surgery, was introduced to avoid ocular complications related to internal limiting membrane peeling, inadequate removal of the vitreous, and incomplete removal of epiretinal membranes. Since 2000, chromovitrectomy has become a popular approach among vitreoretinal specialists. The first vital dye used in chromovitrectomy, indocyanine green, facilitated identification of the fine and transparent internal limiting membrane. Following indocyanine green, trypan blue was introduced to identify epiretinal membranes, and triamcinolone acetonide stained the vitreous well. Recently, additional natural dyes such as lutein and anthocyanin from the açaí fruit have been proposed for intraocular application during vitrectomy. The main goal of this review was to study the role of vital stains in chromovitrectomy and report the latest findings in the literature.
Topics: Coloring Agents; Epiretinal Membrane; Humans; Indocyanine Green; Ophthalmologic Surgical Procedures; Staining and Labeling; Trypan Blue; Vitrectomy; Vitreoretinal Surgery; Vitreous Body
PubMed: 33252365
DOI: 10.1097/APO.0000000000000344 -
Bioscience Reports Jan 2019Staining with Congo Red (CR) is a qualitative method used for the identification of amyloids and in tissue sections. However, the drawbacks and artefacts obtained when... (Review)
Review
Staining with Congo Red (CR) is a qualitative method used for the identification of amyloids and in tissue sections. However, the drawbacks and artefacts obtained when using this dye can be found both and Analysis of scientific data from previous studies shows that CR staining alone is not sufficient for confirmation of the amyloid nature of protein aggregates or for diagnosis of amyloidosis in tissue sections. In the present paper, we describe the characteristics and limitations of other methods used for amyloid studies. Our historical review on the use of CR staining for amyloid studies may provide insight into the pitfalls and caveats related to this technique for researchers considering using this dye.
Topics: Amyloid; Amyloidosis; Benzothiazoles; Coloring Agents; Congo Red; History, 17th Century; History, 18th Century; History, 19th Century; History, 20th Century; History, 21st Century; Humans; Immunohistochemistry; Protein Aggregates; Staining and Labeling
PubMed: 30567726
DOI: 10.1042/BSR20181415 -
Cytometry. Part a : the Journal of the... Dec 2022The progress of digital pathology in recent years has been an opportunity for the development of automated image analysis algorithms for quantitative measurements and...
The progress of digital pathology in recent years has been an opportunity for the development of automated image analysis algorithms for quantitative measurements and computer aided diagnosis. With those new methods comes the need for high staining quality and reproducibility, as image analysis tools are typically more sensible to slight stain variations than trained pathologists. This article presents a method for the automated analysis of cytology slides stains specifically adapted to the challenges encountered in digital cytopathology. In particular, the variety of cell types in cytology slides, the 3D distribution of the cellular material, the presence of superposed cells and the need for independent analysis of sub-cellular compartments are addressed. The proposed method is applied to the quantification of staining variations for quality control, resulting from changes in the staining protocol such as reagent immersion time or a reagent change. Another demonstrated application is the selection of staining protocol parameters that maximize the visible details in nucleus. Finally the analysis pipeline is also used to compare different stain normalization algorithms on digital cytology slides. Code available at: https://gitlab.com/vitadx/articles/automated_staining_analysis.
Topics: Reproducibility of Results; Staining and Labeling; Image Processing, Computer-Assisted; Algorithms; Cytodiagnosis; Coloring Agents
PubMed: 35614552
DOI: 10.1002/cyto.a.24659 -
Histochemistry and Cell Biology Aug 2022Histological slides are an important tool in the diagnosis of tumors as well as of other diseases that affect cell shapes and distributions. Until now, the research...
Histological slides are an important tool in the diagnosis of tumors as well as of other diseases that affect cell shapes and distributions. Until now, the research concerning an optimal staining time has been mainly done empirically. In experimental investigations, it is often not possible to stain an already-stained slide with another stain to receive further information. To overcome these challenges, in the present paper a continuum-based model was developed for conducting a virtual (re-)staining of a scanned histological slide. This model is capable of simulating the staining of cell nuclei with the dye hematoxylin (C.I. 75,290). The transport and binding of the dye are modeled (i) along with the resulting RGB intensities (ii). For (i), a coupled diffusion-reaction equation is used and for (ii) Beer-Lambert's law. For the spatial discretization an approach based on the finite element method (FEM) is used and for the time discretization a finite difference method (FDM). For the validation of the proposed model, frozen sections from human liver biopsies stained with hemalum were used. The staining times were varied so that the development of the staining intensity could be observed over time. The results show that the model is capable of predicting the staining process. The model can therefore be used to perform a virtual (re-)staining of a histological sample. This allows a change of the staining parameters without the need of acquiring an additional sample. The virtual standardization of the staining is the first step towards universal cross-site comparability of histological slides.
Topics: Coloring Agents; Hematoxylin; Humans; Staining and Labeling
PubMed: 35666313
DOI: 10.1007/s00418-022-02118-9