For analysis of tissue quality, samples of VE and PP were taken after 10 h of incubation with or without TNF. Cross-sectioning, deparaffinization, and rehydration were carried out prior to staining. Samples were stained for 3 min with hematoxylin solution according to Harris (Carl Roth GmbH, Karlsruhe, Germany), rinsed in 0.1% hydrochloric acid, and differentiated under flowing water for 3–5 min. The sections were then stained with eosin Y solution (1%; Carl Roth GmbH, Karlsruhe, Germany) for another 3 min, washed with water, and again transferred into increasing alcohol concentrations with a final step of xylol as explained above. Slides were subsequently embedded with ProTaqs paramount (Biocyc, Luckenwalde, Germany).
Hematoxylin solution
Manufactured by Carl Roth
Sourced in Germany
Hematoxylin solution is a laboratory reagent used in various staining procedures, particularly in histology and cytology. It serves as a nuclear stain, highlighting the nuclei of cells. The solution contains hematoxylin, a natural dye derived from the heartwood of the Logwood tree, and other chemical components that facilitate the staining process. This solution is commonly used in combination with other stains to provide contrast and visualization of cellular structures under a microscope.
Automatically generated - may contain errors
Lab products found in correlation
Eosin solution, by Carl Roth (2 mentions)
Eosin y solution, by Carl Roth (1 mentions)
Entellan, by Merck Group (1 mentions)
Zen 2 blue edition software, by Zeiss (1 mentions)
Bouin s, by Merck Group (1 mentions)
Celestine blue solution, by Merck Group (1 mentions)
Axiocam mrm camera, by Zeiss (1 mentions)
Axiovert s100 microscope, by Zeiss (1 mentions)
4 protocols using hematoxylin solution
1
Tissue Quality Analysis via Histological Staining
2
Histopathological Characterization of Healthy and Inflamed Third Molars
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The healthy and inflamed human third molars were histopathologically characterized by hematoxylin and eosin (H&E) staining, as previously described [45 (link)]. The sections were first dried at room temperature for two hours. Subsequently, the sections were washed in distilled water for 5 min and treated with a hematoxylin solution (Carl-Roth GmbH, Karlsruhe, Germany) for 10 min. The sections were washed with slightly warmer running water for 10 min. Following additional washing of the sections with distilled water for 2 min, the sections were treated with eosin solution (Carl-Roth GmbH) for 90 s. The sections were dehydrated in an ascending ethanol series (90%, 95%, 2 × 100% ethanol) for 3 min each. Subsequently, the sections were treated with xylene-I and xylene-II (Carl-Roth GmbH) for 4 min each and coverslipped with Entellan (Merck Millipore, Darmstadt, Germany).
3
Analysis of Ischemia/Reperfusion-Induced Cardiac Fibrosis
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A total of 21 days after ischemia/reperfusion hearts were taken, they were embedded in paraffin and sections of these hearts were prepared. Scar formation was analyzed by staining of these sections with Gomori, Bouin’s (Sigma, Darmstadt, Germany) and hematoxylin solution (Carl Roth, Karlsruhe, Germany). Images were captured by Binocular Microscope (Nikon SMZ25, Tokyo, Japan), evaluated by Zen2 blue edition Software (Zeiss, Oberkochen, Germany) and the ratio of the infarct size to the total of the left ventricle was determined. To determine the amount of interstitial collagen, cardiac sections were stained by Picrosirius red staining (Morphisto, Frankfurt am Main, Germany) and Celestine blue solution (Sigma, Darmstadt, Germany) was used to stain the nuclei. Interstitial collagen was measured in percent by area fraction. Additionally, collagen density was analyzed by polarized light microscopy and evaluated by Image J software.
4
Cryosectioning and Histological Staining
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The cell sheets were fixed in 4% paraformaldehyde/PBS, cryoprotected with sucrose, embedded in tissue freezing medium (Jung, Leica, Nussloch, Baden-Württemberg, Germany) and cryosectioned at 10 μm (Microm HM500 OM, Fisher, Walldorf, Baden-Württemberg, Germany). Sections were stored at −20 °C until use. Before staining, sections were equilibrated to room temperature and hydrated with PBS (3 × 5 min) and rinsed with deionized water. For Haematoxylin and Eosin (H&E), sections were placed in a ready-to-use Hematoxylin solution (Carl Roth, Karlsruhe, Baden-Württemberg, Germany) for 10 min, rinsed with deionized water for 5 min and washed in tap water for 7 min. Next, sections were immersed in a ready-to-use Eosin solution (Carl Roth, Karlsruhe, Baden-Württemberg, Germany) for 10 min, rinsed with distilled water for 30 s, and mounted. Alexa Flour 546-labelled phalloidin (Invitrogen, Karlsruhe, Baden-Württemberg, Germany) was applied according to the manufacturer’s recommendation, as described previously [6 (link)]. DAPI was used for nuclear counterstaining. Cell sheets were imaged with the AxioCamMRm camera on the AxiovertS100 microscope (Carl Zeiss, Jena, Thüringen, Germany). For quantification of the total actin amount, fluorescence signals were quantified using the algorithm described above.
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