• Title/Summary/Keyword: Hematoxylin & Eosin(H&E) staining

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Morphological characteristics of Neural Tissue and Corazonin Neurons of Central Nervous System in Larval Stage of Scuttle Fly

  • Hohyun Park
    • Biomedical Science Letters
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    • v.28 no.4
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    • pp.290-297
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    • 2022
  • Through previous studies, the central nervous system (CNS) was collected by dividing the scuttle fly into larval, pupa, and adult stages by developmental stage, and the morphological characteristics were observed. In situ hybridization (ISH) using the collected central nervous system, it was possible to confirm the location and extent of expression of the neurotransmitter corazonin (Crz) at each stage of development. In this study, paraffin specimens were prepared using central nervous system tissues of 3rd instar larval stage of scuttle fly, which had completed in situ hybridization, and general histochemical staining (hematoxylin-eosin, H-E) and special histochemical staining (luxol fast blue-cresyl violet) was performed to observe the histological and cytological morphology characteristics of corazonin neurons. As a result, a variety of nerve cell body existed between many myelin sheath. The corazonin neurons compose cortex of central nervous system with other neurons congregating in this tissue and show larger cell body relatively in neurohistochemical analysis.

The Novel Label Free Staining Algorithm in Digital Pathology (차세대 디지털 병리를 위한 Label Free 디지털염색 알고리즘 비교 연구)

  • Seok-Min Hwang;Yeun-Woo Jung;Dong-Bum Kim;Seung Ah Lee;Nam Hoon Cho;Jong-Ha Lee
    • Journal of the Institute of Convergence Signal Processing
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    • v.24 no.1
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    • pp.76-81
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    • 2023
  • To distinguish cancer cells from normal cells, H&E (Hematoxylin & Eosin) staining is required. Pathological staining requires a lot of money and time. Recently, a digital dyeing method has been introduced to reduce such cost and time. In this paper, we propose a novel digital pathology algorithms. The first algorithm is the Pair method. This method learns the dyed phase image and unstained amplitude image taken by FPM (Fourier Ptychographic Microscopy) and converts it into a dyed amplitude image. The second algorithm is the unpair method. This method use the stained and unstained fluorescence microscopic images for modeling. In this study, digital staining was performed using a generative adversarial network (GAN). From the experimental results, we noticed that both the pair and unpair algorithms shows the excellent performance.

Evaluating the Efficacy of a Formalin Alternative in Fixing Pathological Tissues for Histological and Molecular Diagnoses

  • Min-A Je;Haneul Lee;Heechul Park;Dong Hyeok Kim;Yeongdon Ju;Jaewon Lim;Sunghyun Kim;Jungho Kim
    • Biomedical Science Letters
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    • v.29 no.1
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    • pp.48-52
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    • 2023
  • Formaldehyde use is associated with serious health risks, which can affect medical personnel and technicians. Therefore, we investigated the efficacy of an alternative fixative, with respect to two types of formalin fixatives, by hematoxylin and eosin (H&E) staining, periodic acid Schiff (PAS) staining, immunohistochemical (IHC) staining, and RNA extraction. For H&E staining, the circular nucleus was stained dark blue by the basic dye hematoxylin and the cytoplasm was stained red by the acid dye eosin in all three fixative samples. No difference was found in the Duksan General Science (DGS), Sigma-Aldrich, and Core-Fix fixative samples (Corebiotech) used to fix kidney tissue, after PAS staining. IHC staining showed that CD4 was significantly increased in the lippolysaccharide (LPS)-treated group compared to the control group (vehicle), confirming the changes in specific molecules. The quantity and quality of RNA from tissues fixed in the three types of fixatives were evaluated. The average concentration of RNA was 106 ng/µL and average purity at A 260/280 ratio was 1.7~2.0, regardless of fixative used. For quality of protein, glyceraldehyde 3-phosphate dehydrogenase (GAPDH) protein was confirmed by Western blotting. In conclusion, Core-Fix can be used as a fixative for pathological tissues, in histological and molecular diagnoses.

Comparative Analysis of Histochemical Stains about Detection of H. pylori in Gastric Mucosa (위점막에서 H. pylori 검출을 위한 조직염색법의 비교)

  • Cha, Min-Seon
    • Korean Journal of Clinical Laboratory Science
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    • v.39 no.3
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    • pp.223-230
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    • 2007
  • Several methods have been used in the detection of Helicobacter pylori (H. pylori) which was believed to be a pathogenic organism causing chronic gastritis, benign peptic ulcer, gastric carcinoma or malignant lymphoma. Even though several methods were introduced for detection of H. pylori in stomach, there were controversies in their sensitivities and specificities. This experiment were designed to study the comparative analysis of staining methods (hematoxylin and eosin (H&E), Giemsa, Warthin-Starry and immunohistochemical stain) to dectect H. pylori in the gastric mucosa. The results were as follows. Average density score of H. pylori classified by Genta were 2.29 in Warthin-Starry stain, 2.19 in Giemsa stain, 1.34 in immunohistochemical stain and 0.98 in H&E stain. By comparison between inflammatory degree by Sydney system and result of Warthin-Starry stain, the detection rate and densities of H. pylori were increased from mild (61.5% and 0.8), moderate (90.4 and 2.1), and severe (100% and 3.2). From the above findings, Warthin-Starry stain is useful method for detection of H. pylori in gastric mucosa.

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A Study of the Histological Effects of Garlic Extract in a Liver-damage Rat Model (마늘 추출물이 간 손상 랫드의 조직학적 변화에 미치는 효과 연구)

  • Lee, Tae-Jong;Min, Kyung-Jin
    • Journal of Environmental Health Sciences
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    • v.36 no.4
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    • pp.279-287
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    • 2010
  • In this study we examined the effects of water extract of garlic on carbon tetrachloride-induced liver injury, and demonstrated increased beneficial enzyme and anti-oxidant activity as well as histopathological changes (by Hematoxylin-Eosin (H&E) staining, Trichrome staining, and TEM examination), and showed that the treatment was dose-dependent and safe. A total of 42 male Sprague-Dawley rats were divided equally (n=7) into six groups. To induce hepatotoxicity in these subjects, carbon tetrachloride diluted in an equal volume of olive oil was intraperitoneally administrated at 0.5 ml/kg (0.20 g/kg/day) once a day for five days. Water extract of Korean-grown garlic was administered via a stomach sonde once a day, 5 days a week, for a total of 4 weeks. Groups received 0.35 g/kg (E1), 0.70 g/kg (E2), or 1.40 g/kg (E3), with the dose adjusted for body weight. Administration of garlic extract resulted in positive physiological effects in terms of reduced oxidative stress and toxicity, and induced functional changes in the liver. Comparing the subject groups (E1, E2, E3) administered different doses of garlic extract, the importance of morphological analysis in further studies is emphasized, because morphological changes indicating hepatotoxicity could occur, even though beneficial enzyme activities were found to be elevated.

Morphological changes by whole-body r -irradiation in mouse jejunal villi (생쥐 공장 융모의 감마선 전신조사에 의한 형태학적 변화)

  • Chun, Ki-Jung;Kim, Jin-Kyu;Song, Chi-Won;Kim, Moo-Kang
    • Journal of Radiation Protection and Research
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    • v.25 no.4
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    • pp.217-221
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    • 2000
  • Radiotheraphy may be considered as one of the most effective treatments for digestive tumors. This procedure has major side effects, especially in fast growing tissues like intestinal mucosa. For this reason, the morphological changes in mouse jejunal villi after whole body ${\gamma}-irradiation$ were analysed. Four week old ICR male mice were irradiatied with 6.5Gy or 12Gy of whole body ${\gamma}-irradiation$ and were sacrificed 3 days later. Jejunum among intestine were taken for morphology. Samples were carried out dehydration process and sliced sample covered with paraffin was hold on the slide and then light microscopy was observed by Hematoxylin & Eosin staining. villi at both irradiated doses were showed that the length of villi were shortened and thickened and that lumen were expanded in comparison with non-irradiated group. Since willi have an important role in digestion and very sensitive to radiation from this test, it has a role of test parameter for finding radioprotectors as well as evaluating the biological effect by radiaton.

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Modified Toluidine Blue: an Alternative Stain for Helicobacter pylori Detection in Routine Diagnostic Use and Post-eradication Confirmation for Gastric Cancer Prevention

  • Sakonlaya, Dussadee;Apisarnthanarak, Anucha;Yamada, Nobutaka;Tomtitchong, Prakitpunthu
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.16
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    • pp.6983-6987
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    • 2014
  • Background: Modified toluidine blue staining (MTBs) is a simple, inexpensive and time saving method to detect H. pylori in gastric biopsy specimens. As a metachromatic stain, it simultaneously highlights intestinal metaplasia, a gastric cancer precancerous lesion. The aim of this study was to assess the reliability of MTBs compared with hematoxylin-eosin (H&E) for H. pylori detection using immunoperoxidase staining as the gold standard. This technique would be beneficial for a routine diagnosis and confirmation of H. pylori eradication in developing countries where endoscopic-based approaches are dominant. Materials and Methods: Esophagogastroduodenoscopy with triple site gastric biopsies was undertaken in 207 dyspeptic patients at Thammasat University Hospital, Thailand between 1997 and 1999. H&E, MTBs and immunoperoxidase staining were applied to each specimen. The presence or absence of H. pylori with each stain was interpreted separately and the sensitivity, specificity, positive and negative predictive values of H&E and MTBs were calculated. Results: A total of 282 specimens from 207 patients were evaluated. Using immunoperoxidase staining, organisms were positive in 117 specimens (41%). MTBs proved almost equally sensitive as immunoperoxidase (99%) and significantly more sensitive than H&E (85%). It has comparable specificity (96% vs 96%), PPV (95% vs 94%), and NPV (99% vs 90%) to H&E, using immunoperoxidase staining as gold standard. MTBs compared with immunoperoxidase staining, is cheaper (2 USD vs 12 USD) and faster (20 min vs 16 hrs) compared to immunoperoxidase staining. Conclusions: MTBs is effective, economical and easy to use in daily practice for the detection of H. pylori in gastric biopsy specimens. In addition to saving time in evaluating H. pylori associated gastritis, with a high sensitivity and ability to demonstrate intestinal metaplasia, the technique may have a role in confirmation of H. pylori eradication for gastric cancer prevention in a developing country setting.

Quantitative Analyses of Cells using Photoshop after the H&E Staining of the Synovia of Osteoarthritis and Rheumatoid Arthritis Patients (H&E 염색 이미지의 포토샵 분석을 이용한 골관절염과 류마티스 관절염 활막 세포의 정량 분석)

  • Park, Jin-Ah;Kim, Keun-Cheol
    • Journal of Life Science
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    • v.22 no.8
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    • pp.1034-1040
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    • 2012
  • Synovium is the soft tissue that lines the non-cartilaginous surfaces within joints. It has been reported that synovial cells are activated during the pathogenesis of rheumatoid arthritis. In this study, we quantitate and compare the cellular composition of synovia derived from individuals with non-inflammatory osteoarthritis (OA) and those with inflammatory rheumatoid arthritis (RA). Synovia from OA (n=8) and RA (n=5) patients were used for hematoxylin and eosin (H&E) staining. A light microscopic examination has shown that RA synovia were morphologically thickened and hypertrophied as compared to OA synovia. We also performed an immunohistochemistry (IHC) analysis to classify cell types in the synovia using CD68, CD90, or PGP9.5 markers. As a result, we obtained quantitative data regarding the cell populations, which are macrophages in the lining layer and FLSs in the subintimal layer of the synovium. Further Photoshop analyses of the H&E images could allow the counting of the number and layer of the cells in the synovium. The number and layers of the macrophage cells were increased in the lining layer of the RA synovia as compared to the OA synovia. FLS cells also were increased in the subintimal layer of RA synovia. Therefore, quantification of the H&E stained images via Photoshop is a possible analysis protocol for synovium study. This quantitation also supports the idea that the increases in cell number and cell activation are important processes for RA pathogenesis.

Use of Anti-Phosphohistone H3 (PHH3) as a Mitosis Marker for Classifying Pulmonary Carcinoid Tumors

  • Seo, Bo-Ram;Hong, Young-Seob;Choi, Phil-Jo;Um, Soo-Jung;Seo, Jeong-Wook;Roh, Mee-Sook
    • Biomedical Science Letters
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    • v.17 no.3
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    • pp.197-202
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    • 2011
  • Mitosis count is one of the most helpful morphologic features for distinguishing pulmonary typical carcinoid (TC) from atypical carcinoid (AC). However, identifying areas of highest mitotic activity is tedious and time-consuming, and mitosis count may vary substantially among pathologists. Anti-phosphohistone H3 (PHH3) is an antibody that specifically detects histone H3 only when phosphorylated at serine 10 or serine 28, an event that is concurrent with mitotic chromatin condensation and not observed during apoptosis. In this study, immunohistochemical staining for PHH3 was performed to determine whether PHH3 was a reliable and objective mitosis-specific marker for pulmonary carcinoid tumors. Seventeen cases of surgically resected pulmonary carcinoid tumors (12 TCs and 5 ACs) were obtained and classified according to the 2004 World Health Organization classification. Mitotic counts determined by PHH3 correlated to ones determined by hematoxylin and eosin (H&E) staining; however, PHH3 mitotic counts (mean mitotic counts: 1 in TCs and 3.2 in ACs) were slightly higher than H&E mitotic counts (mean mitotic counts: 0.25 in TCs and 1.8 in ACs). The mitotic counts determined by experienced observer were more correlated to those determined by inexperienced observer with the PHH3-based method (R=0.968, P<0.001) rather than H&E staining (R=0.658, P<0.001). These results suggest that the PHH3 mitotic counting method was more sensitive and simple for detecting mitoses compared to traditional H&E staining. Therefore, PHH3 immunohistochemistry may contribute to more accurate and reproducible diagnosis of pulmonary carcinoid tumors and may be a valuable aid for administrating appropriate clinical treatment.

Development of Cryptosporidium parvum in cell culture (세포배양에서 Cryptosporidium parvum의 발육)

  • Kim, Bo-sook;Joo, Hoo-don;Wee, Sung-hwan;Kim, Tae-jong
    • Korean Journal of Veterinary Research
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    • v.35 no.2
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    • pp.317-326
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    • 1995
  • The purpose of this study was to establish a method for in vitro culture of C parvum isolated in Korea by determination of suitable cell model to complete development of this parasite. The result obtained were summerized as follows: 1. To determine the most suitable cell line, six types of cell line were examined by microscopy. All cell lines were infected with C parvum and showed the highest infection score in HmLu cells. 2. The staining methods including DMSO-modified acid-fast(A-F) stain, hematoxylin-eosin(H & E) stain and immunofluorescence antibody(IFA) stain were applied to examine the infection of C parvum in cell culture. These staining methods were possible to examine the infection of C parvum in cell culture. The most sensitive one was IFA staining technique. 3. Developmental stages of C parvum in HmLu cell were observed. After the initial 8 hour incubation period, some trophozoites were observed. The meronts and gametes were appeared at 24-48 hour post inoculation(PI), and oocysts were observed firstly at 48-72 hour PI. 4. In H & E stain, the parasite appeared as basophilic within parasitophorous vacuole membrane(PVM) and lying in cytoplasm at near the nucleus of the host cells. It was able to distinguish the type I, type II meronts and gametes. 5. In DMSO-modified acid-fast stain, specific stained parasites were appeared firstly after 48 hour PI. The parasites were showed with different degrees of staining bright red color within PVM. 6. The endogenous stages of parasites in HmLu cell recovered at 48, 96, 120 and 144 hour after inoculation were reacted with rabbit immunized serum in immunofluorescence antibody and avidin-biotin complex peroxidase staining technique.

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