After deparaffination and dehydration, the slides were stained with Mayer‘s hemalum solution (Merck, USA) for three minutes and were differentiated under running tap water. Afterward, they were stained for 3 min with 0.5% Eosin G solution (Roth, Germany) enriched with glacial acetic acid (Roth, Germany).
Eosin g solution
Manufactured by Carl Roth
Sourced in Germany
Eosin-G solution is a laboratory reagent used for various applications in the life sciences. It is a synthetic dye that exhibits a bright pink-red color. Eosin-G solution is commonly used as a counterstain in histological and cytological staining procedures.
Automatically generated - may contain errors
Lab products found in correlation
Agarose, by Merck Group (2 mentions)
Mayer s hemalum solution, by Merck Group (1 mentions)
Glacial acetic acid, by Carl Roth (1 mentions)
Tissue tek oct compound, by Sakura Finetek (1 mentions)
Safranin o solution, by Merck Group (1 mentions)
Fast green solution, by Merck Group (1 mentions)
Mayer s haematoxylin, by Agilent Technologies (1 mentions)
Nuclear fast red aluminium sulphate solution, by Carl Roth (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
4 protocols using eosin g solution
1
Hematoxylin and Eosin Staining Protocol
2
Histological Evaluation of Intervertebral Disc Components
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To confirm correct separation of AF and NP in macroscopic preparation, we performed standard histological staining of each AF and NP sample. A representative small tissue part was embedded in Tissue-Tek® O.C.T. Compound (Sakura Finetek, Staufen, Germany), frozen in liquid nitrogen and stored at − 80 °C until sectioning. Samples were cryosectioned at a thickness of 6 μm. To show tissue morphology, sections were fixed with methanol/acetone (1:1 v/v %) and stained with Mayer’s haematoxylin (Dako, Germany) and eosin-G solution (Roth, Karlsruhe, Germany). To analyse sulphated GAGs, the sections were fixed with 4% formaldehyde (Herbeta, Germany) and stained with 1% alcian blue 8GS solution (Roth) and counterstained with nuclear fast red-aluminium sulphate solution (Roth). To demonstrate acidic GAGs, the sections were fixed with 92% ethanol and stained with 0.7% safranin O solution (Sigma-Aldrich) and subsequently counterstained with 0.2% fast green solution (Sigma-Aldrich).
3
Cisplatin-Induced Apoptosis Cytometry Protocol
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Cells were cultured in T-175 flasks (4 × 106 cells/flask) and treated in the same manner as with the cells being analyzed using the ApoTox-Glo® Triplex assay. After 48 h of incubation from cisplatin treatment, cells were harvested by incubating with 0.25% Trypsin–EDTA (Thermo Fisher Scientific) for 2–5 min, centrifuged for 3 min at 300×g and then fixed in 8 ml of 4.5% Formalin (Otto Fischar GmbH & Co. KG, Saarbrucken, Germany).
Fixed cells were centrifuged at 800×g for 10 min. The cell pellet was transferred into a 1.5 ml reaction tube and stained with one drop of Eosin-G solution (0.5%, Carl Roth, Karlsruhe, Germany). The tubes were centrifuged at 4000×g for 2 min. The supernatant was removed by a pipette. The pellet was mixed with ca. 1 ml of 1% agarose (Merck KGaA, Darmstadt, Germany). After solidification (ca. 10 min), the gel was bisected and transferred into embedding cassettes. Cassettes were stored in 4.5% Formalin until dehydration. After dehydration, cassettes were embedded in paraffin and stored at room temperature.
Fixed cells were centrifuged at 800×g for 10 min. The cell pellet was transferred into a 1.5 ml reaction tube and stained with one drop of Eosin-G solution (0.5%, Carl Roth, Karlsruhe, Germany). The tubes were centrifuged at 4000×g for 2 min. The supernatant was removed by a pipette. The pellet was mixed with ca. 1 ml of 1% agarose (Merck KGaA, Darmstadt, Germany). After solidification (ca. 10 min), the gel was bisected and transferred into embedding cassettes. Cassettes were stored in 4.5% Formalin until dehydration. After dehydration, cassettes were embedded in paraffin and stored at room temperature.
4
FFPE-based quantitative PCR for MT gene expression
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The cells were embedded in FFPE to assess gene expression of MTs via quantitative PCR. As FFPE is the most commonly available form of patient material, we performed this step, instead of isolating RNA directly from the treated cells. Therefore, cells were seeded in flasks (4 million per flask) and incubated with or without different concentrations of zinc (10-, 40-, 70 µM) for either 48- or 77 h at 37 ℃, 5% CO2. After incubation, cells were harvested. The cells were fixed in 4.5% formalin (Otto Fischar GmbH & Co. KG, Saarbrücken, Germany) and embedded into paraffin blocks. The cells, fixed in formalin, were centrifuged at 800×g for 10 min. The pellet was moved into a 1.5 mL Eppendorf tube and staining with Eosin-G solution was performed (0.5%, Carl Roth, Karlsruhe, Germany). Subsequently, the tubes were centrifuged at 4,000×g for 2 min. The supernatant was removed by using a Pasteur pipette. The pellet was then mixed by using 1 mL of 1% agarose (Merck KGaA, Darmstadt, Germany). After solidification (ca. 10 min), the gel was divided and moved into embedding cassettes. These cassettes were stored in 4.5% buffered formalin until the dehydration process. Cassettes were embedded in paraffin after dehydration and then stored at room temperature for further analyses.
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