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Hematoxylin eosin

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Hematoxylin & Eosin is a staining solution used in histological and pathological laboratories. It is a combination of two dyes, hematoxylin and eosin, that stain different cellular components. Hematoxylin stains nuclei blue, while eosin stains cytoplasm and other structures pink or red. This staining technique is a widely used method for the visualization and examination of tissue samples.

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Lab products found in correlation

Bond polymer refine detection, by Leica Biosystems (3 mentions) Bond rxm, by Leica Biosystems (3 mentions) Bond intense r detection systems, by Leica Biosystems (3 mentions) Picrosirius red, by Merck Group (2 mentions) Vectamount, by Vector Laboratories (2 mentions) Masson s trichrome, by Diagnostic BioSystems (2 mentions) Anti bcl6, by Santa Cruz Biotechnology (2 mentions) Ra3 6b2, by BD (2 mentions) Fisher brand superfrost plus glass slides, by Thermo Fisher Scientific (2 mentions) Eclipse 50i, by Nikon (1 mentions) Sodium hydroxide (naoh), by Thermo Fisher Scientific (1 mentions) S7110, by Merck Group (1 mentions) Bromodeoxyuridine (brdu), by Abcam (1 mentions) P stat1, by Cell Signaling Technology (1 mentions) γh2ax, by Cell Signaling Technology (1 mentions) Cytoseal, by Thermo Fisher Scientific (1 mentions) Podoplanin, by BioLegend (1 mentions) Foxp3, by Thermo Fisher Scientific (1 mentions) Cytokeratin 17 19, by Cell Signaling Technology (1 mentions) Anti b220, by BD (1 mentions)

18 protocols using hematoxylin eosin

1

Asbestos Body Identification in Lung Tissue

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A light microscope (ECLIPSE 50i; Nikon Corp., Tokyo, Japan) was used to examine and count the presence of asbestos bodies in specimens of lung tissue or bronchoalveolar lavage fluid. The lung tissue specimens were stained using hematoxylin-eosin (Thermo Fisher Scientific, Inc.). The fluid specimens were stained using the papanicolaou method (19 ) (Fig. 2), subsequent to breaking down the mucus in the fluid using 10% NaOH (Thermo Fisher Scientific, Inc.). A specimen with >5 asbestos bodies in every 10 examined fields with a ×40 magnification was considered to contain asbestos bodies (20 (link)).
THANH T.D., THO N.V., LAM N.S., DUNG N.H., TABATA C, & NAKANO Y. (2016). Simian virus 40 may be associated with developing malignant pleural mesothelioma. Oncology Letters, 11(3), 2051-2056.
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2

Histological Analysis of Tissue Samples

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Histological and immunohistochemistry studies were performed using formalin‐fixed paraffin‐embedded 3‐μm tissue sections. Signs of histological injury were examined with hematoxylin/eosin (Cat#10034813 and Cat#6766007, ThermoFisher) or periodic acid‐Schiff (PAS) staining (Cat#3952016, Sigma‐Aldrich). Full methods for F4/80 immunohistochemistry and p65 immunofluorescence are presented in the Supplementary materials and methods. The presence of cell death was determined using a terminal deoxynucleotidyl transferase dUTP nick‐end labeling (TUNEL) assay following the manufacturer's instruction (Cat#S7110, Millipore, Burlington, MA, USA).
Vázquez‐Carballo C., Herencia C., Guerrero‐Hue M., García‐Caballero C., Rayego‐Mateos S., Morgado‐Pascual J.L., Opazo‐Rios L., González‐Guerrero C., Vallejo‐Mudarra M., Cortegano I., Gaspar M.L., de Andrés B., Egido J, & Moreno J.A. (2022). Role of Toll‐like receptor 4 in intravascular hemolysis‐mediated injury. The Journal of Pathology, 258(3), 236-249.
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3

Comprehensive Immunohistochemical Analysis of Tissue Samples

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Tissues were fixed in 10% neutral formalin for 18–24 hours, embedded in paraffin after graded-ethanol dehydration, and sectioned into 5-μm sections using a microtome. Where applicable, formalin-fixed, paraffin embedded (FFPE) tissue sections were stained for Hematoxylin & Eosin (Thermo Fisher), Picro-Sirius Red (Sigma-Aldrich) and Masson’s Trichrome (Diagnostic Biosystems) according to manufacturer’s instructions. Automated staining of tissues was carried out on the Bond RXm (Leica Biosystems) following dewaxing and appropriate antigen retrieval. Immunostaining was chromogenically visualized using the Bond Polymer Refine Detection alone or in conjunction with Bond Intense R Detection Systems (DS9263, Leica Biosystems). Slides were dehydrated through graded ethanol, followed by xylene, then mounted using Xylene-based Cytoseal (Thermo Fisher) or Vectamount (Vector Labs) as appropriate.
Staining was performed with the following antibodies: αSMA (Abcam ab5694), Podoplanin (Biolegend 127402), Ki67 (Abcam ab15580), BrdU (Abcam ab2284), CC3 (Cell Signaling 9661S), Cytokeratin 19 (DSHB TROMA-III), Cytokeratin 17/19 (Cell Signaling 12434S), CD8α (Cell Signaling 98941), pSTAT1 (Cell Signaling 8826s), CD4 (Abcam ab183685), Foxp3 (eBioscience 14–5773-82), γH2Ax (Cell Signaling 9718S).
Lander V.E., Belle J.I., Kingstonl N.L., Herndon J.M., Hogg G.D., Liu X., Kang L.I., Knolhoff B.L., Bogner S.J., Baer J.M., Zuo C., Borcherding N.C., Lander D.P., Mpoy C., Scott J., Zahner M., Rogers B.E., Schwarz J.K., Kim H, & DeNardo D.G. (2022). Stromal reprogramming by FAK inhibition overcomes radiation resistance to allow for immune priming and response to checkpoint blockade. Cancer discovery, 12(12), 2774-2799.
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4

Histological Tissue Analysis Protocols

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Tissues were fixed in formalin for 24 h, embedded in paraffin after graded-ethanol dehydration, and sectioned into 5-μm sections using a microtome. FFPE sections were stained for Hematoxylin & Eosin (Thermo Fisher) and Modified Masson’s Trichrome (Diagnostic Biosystems KT034) according to manufacturer’s instructions. After dewaxing and epitope retrieval, tissues were auto-stained on the Bond Rxm (Leica Biosystems). Staining was visualized using the Bond Polymer Refine Detection alone or in conjunction with Bond Intense R Detection Systems (DS9263, Leica Biosystems). Antibodies used are in Supplemental Table 1. Additional details are in supplemental methods.
Akinjiyan F.A., Ibitoye Z., Zhao P., Shriver L.P., Patti G.J., Longmore G.D, & Fuh K.C. (2023). DDR2-regulated arginase activity in ovarian cancer-associated fibroblasts promotes collagen production and tumor progression. Oncogene, 43(3), 189-201.
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5

Histological and Immunohistochemical Analysis of Mouse Organs

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Histological analysis of mouse organs was performed on 4µm-thick
FFPE tissue sections, stained with Hematoxylin & Eosin (Thermo
Scientific) following standard procedures. The following primary antibodies were
used for immunohistochemical analysis: anti-Bcl6 (1:300) (N3, rabbit polyclonal,
Santa Cruz Biotechnology), anti-PAX5 (1:400) (rabbit polyclonal, Neomarker),
anti-CD3 (1:800) (rabbit monoclonal, clone SP7, Neomarker), anti-B220 (1:400)
(rat monoclonal, clone RA3-6B2, BD Biosciences), anti-IRF4 (1:200) (M-17, goat
polyclonal, Santa Cruz Biotechnology), anti-BCL2 human (1:100) (rabbit
polyclonal, Santa Cruz Biotechnology), and anti-CD138 (1:200) (rat monoclonal,
clone 281-2, BD Biosciences).
Zhang J., Vlasevska S., Wells V.A., Nataraj S., Holmes A.B., Duval R., Meyer S.N., Mo T., Basso K., Brindle P.K., Hussein S., Dalla-Favera R, & Pasqualucci L. (2017). The Crebbp acetyltransferase is a haploinsufficient tumor suppressor in B cell lymphoma. Cancer discovery, 7(3), 322-337.
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6

Histological Analysis of Mouse Lymphoid Organs

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Histological analysis of mouse lymphoid organs was performed on
3-μm-thick FFPE tissue sections, stained with Hematoxylin & Eosin
(Thermo Scientific) following standard procedures. The following primary
antibodies were used for immunohistochemical analysis: anti-Bcl6 (1:300)
(N3, rabbit polyclonal, Santa Cruz Biotechnology); biotin-conjugated
anti-PNA (1:200) (Vector Laboratories); biotin-conjugated anti-B220 (1:400)
(RA3–6B2, rat monoclonal, Pharmingen 553086) and anti-CD3 (1:800)
(SP7, rabbit monoclonal, NeoMarkers RM9107). GC numbers, size, and overall
area were calculated using the ImageJ software on scanned images obtained
with a Leica SCN400 slide scanner (Schindelin et al., 2015 (link)).
Meyer S.N., Scuoppo C., Vlasevska S., Bal E., Holmes A.B., Holloman M., Garcia-Ibanez L., Nataraj S., Duval R., Vantrimpont T., Basso K., Brooks N., Dalla-Favera R, & Pasqualucci L. (2019). Unique and Shared Epigenetic Programs of the CREBBP and EP300 Acetyltransferases in Germinal Center B Cells Reveal Targetable Dependencies in Lymphoma. Immunity, 51(3), 535-547.e9.
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7

Tissue Histological Analysis Protocol

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Tissues were fixed in 10% neutral-formalin for 18 hours, embedded in paraffin after graded-ethanol dehydration, and sectioned into 6-μm sections using a microtome. Where applicable, FFPE sections were stained for Hematoxylin & Eosin (Thermo Fisher), Picro-Sirius Red (Sigma-Aldrich) and Masson’s Trichrome (Diagnostic Biosystems) according to manufacturer’s instructions. Automated staining of tissues was carried out on the Bond Rxm (Leica Biosystems) following dewaxing and appropriate epitope retrieval. Immunostaining was chromogenically visualized using the Bond Polymer Refine Detection alone or in conjunction with Bond Intense R Detection Systems (DS9263, Leica Biosystems). Staining used antibodies listed in Key Resources Table. Slides were mounted using Xylene-based Cytoseal (Thermo Fisher) or Vectamount (Vector Labs) as appropriate.
Hegde S., Krisnawan V.E., Herzog B.H., Zuo C., Breden M.A., Knolhoff B.L., Hogg G.D., Tang J.P., Baer J.M., Mpoy C., Lee K.B., Alexander K.A., Rogers B.E., Murphy K.M., Hawkins W.G., Fields R.C., DeSelm C.J., Schwarz J.K, & DeNardo D.G. (2020). Dendritic cell paucity leads to dysfunctional immune surveillance in pancreatic cancer. Cancer cell, 37(3), 289-307.e9.
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8

Histological Analysis of Murine Muscle

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Male mice at various ages (indicated where appropriate) were euthanized via CO2 asphyxiation and cervical dislocation. Hind limb muscles were dissected and fixed for 24 h with 10% phosphate-buffered formalin, followed by a 70% ethanol incubation for 24 h. Tissue was then processed and embedded in paraffin (Thermo Scientific, Waltham, MA, USA) and 12 μm sections were collected on SuperFrost Plus slides (Fisher Scientific, Hampton, NH, USA). Slides were deparaffinized with two xylene incubations and rehydrated with a series of ethanol incubations at the following concentrations: 100%, 100%, 95%, 70%, 50%. The slides were then stained for hematoxylin/eosin (Thermo Scientific, Waltham, MA, USA) or Masson’s trichrome (American MasterTech, Lodi, CA, USA) for immunohistochemical staining and analysis.
Paleo B.J., McElhanon K.E., Bulgart H.R., Banford K.K., Beck E.X., Sattler K.M., Goines B.N., Ratcliff S.L., Crowe K.E, & Weisleder N. (2022). Reduced Sarcolemmal Membrane Repair Exacerbates Striated Muscle Pathology in a Mouse Model of Duchenne Muscular Dystrophy. Cells, 11(9), 1417.
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9

Chitosan-based Hydrogel for Chondrocyte Culture

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Chitosan (degree of deacetylation ~ 95%, viscosity 100–200 mPa.s) was obtained from Aladdin (Shanghai, China). Sodium monoiodoacetate (MIA) and sodium periodate (NaIO4) were supplied by Sigma-Aldrich (St. Louis, MO, USA). Enzyme-linked immunosorbent assay (ELISA) kit of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, and IL-17 were purchased from Sigma-Aldrich. Hyaluronic acid (100–200 k) and adipic acid dihydrazide (ADH) were produced by Yuanye biology (Shanghai, China). Rat chondrocytes (ATCC-0038) was got from American Type Culture Collection (ATCC, Gaithersburg, MD, USA). Dulbecco’s Modified Eagle’s Medium (DMEM, low glucose), streptomycin–penicillin and fetal bovine serum (FBS) were supplied by Gibco® Life Technologies (Carlsbad, CA, USA). Cell Counting Kit-8 (CCK-8) and Calcein-AM/Propidium Iodide (PI) were purchased from Beyotime Biotechnology (Shanghai, China). Hematoxylin–Eosin and Safranin O stains were got from Thermo Fisher Scientific Co., Ltd (Shanghai, China). Triton X-100 was supplied by Solarbio Science & Technology Co., Ltd. (Beijing, China). The antibodies enrolled in immunofluorescence were obtained from Abcam (Cambridge, UK).
Mou D., Yu Q., Zhang J., Zhou J., Li X., Zhuang W, & Yang X. (2021). Intra-articular Injection of Chitosan-Based Supramolecular Hydrogel for Osteoarthritis Treatment. Tissue Engineering and Regenerative Medicine, 18(1), 113-125.
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10

Histological Analysis of Jaw Specimens

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Surgical specimens were taken and fixed by immersion in 10% formaldehyde solution (Sigma-Aldrich Inc.) pH 7.4 at 4°C for 72 hours, decalcified in Shandon TBD-1 Rapid Decalcifer (Thermo Fisher Scientific) for 24 hours, dehydrated in ascending series of ethanol (70, 80, 90 and 100%) and transferred to a solution of xylene 100% for 30 minutes. Samples were embedded in paraffin, and 4µm histological sections were stained with hematoxylin-eosin (Thermo Fisher Scientific), blue toluidine (Thermo Fisher Scientific), to identify condroitin sulfate in the cartilage matrix and intramembranous bone, and Masson trichrome (Thermo Fisher Scientific), to observe collagen fibers and calcification process.
To track Ad-MSC, mandibular specimens were fixed in a solution of 10% formaldehyde, pH 7.4 at 4°C for 72 hours, decalcified in EDTA for two months, dehydrated in ascending series of ethanol and transferred to a solution of xylene 100% for 30 minutes. Samples were embedded in paraffin, and 4 µm histological sections obtained for immunohistochemical detection of human ß-2 microglobulin positive cells.
Linero I, & Chaparro O. (2014). Paracrine Effect of Mesenchymal Stem Cells Derived from Human Adipose Tissue in Bone Regeneration. PLoS ONE, 9(9), e107001.
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