Iron deposition in SH-SY5Y cells of each group was detected with an Iron Stain Kit (Sigma-Aldrich, MO, USA). According to the manufacturer’s protocol, the fixed cells were stained with iron staining solution and then pararosanline solution. Micrographs were taken using an Olympus DP74 microscope (Olympus Co., Tokyo, Japan). The quantitative analysis of the relative iron-stained area was performed using Image J software (Image J 1.4, NIH, USA).
Iron stain kit
Manufactured by Merck Group
Sourced in United States, Germany
The Iron Stain Kit is a laboratory tool used to stain and visualize iron-containing compounds in biological samples. It provides a simple and reliable method for the detection of iron, which is an essential mineral in various cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Primovert, by Zeiss (4 mentions)
Nuclear fast red solution, by Merck Group (2 mentions)
Axioskop 40, by Zeiss (2 mentions)
Entellan, by Merck Group (2 mentions)
Metamorph software, by Molecular Devices (2 mentions)
Aperio at2, by Leica camera (1 mentions)
Ventana benchmark ultra, by Roche (1 mentions)
Microtome, by Reichert Technologies (1 mentions)
Lectin, by Vector Laboratories (1 mentions)
Alexa fluor 488 goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 594 goat anti rat igg, by Thermo Fisher Scientific (1 mentions)
Anti rat igg, by Vector Laboratories (1 mentions)
Vector immunodetection kit, by Vector Laboratories (1 mentions)
Vectastain abc kit, by Vector Laboratories (1 mentions)
Hoechst 33258, by Merck Group (1 mentions)
1 2 dimyristoyl sn glycero 3 phosphocholine, by Avanti Polar Lipids (1 mentions)
1 myristoyl 2 hydroxy sn glycero 3 phosphocholine, by Avanti Polar Lipids (1 mentions)
Tissuefaxs platform, by TissueGnostics (1 mentions)
Moticam 2300, by Nikon (1 mentions)
Diaphot microscope, by Nikon (1 mentions)
31 protocols using iron stain kit
1
Quantifying Iron Deposition in SH-SY5Y Cells
2
Iron Staining Protocol
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Iron staining was performed using an iron stain kit obtained from Sigma-Aldrich Co. (St. Louis, MO, USA) according to the manufacturer's recommendations.
3
Histological Analysis of Liver Biopsies in T2DM
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Formalin-fixed paraffin-embedded liver biopsies from T2DM patients were cut with a thickness of 3 μm. After rehydration, the slides were processed for iron staining or CD68 immunohistochemistry.
Standard Perls Prussian blue staining was performed by using the Iron Stain Kit (Sigma–Aldrich). Briefly, the slides were incubated for 15 min in iron stain solution (prepared by mixing equal volumes of 4% w/v potassium ferrocyanide and 1.2 mM hydrochloric acid). Nuclear Fast Red was used as counterstain.
CD68 expression was analyzed by CD68 antibody staining (Dako; Ref-Nr.:M0876, Lot-Nr.: 00094,759; clone PG-M1) on the Ventana Benchmark Ultra (Roche) according to the manufacturer's instructions. Microscopy slides containing the tissue were preincubated with the EDTA-containing CC1 solution (Roche) for 16 min after which the slides were incubated with a 1:100 dilution of the CD68 antibody for 24 min.
The stained slides were digitally scanned with the Aperio AT2 (Leica).
Standard Perls Prussian blue staining was performed by using the Iron Stain Kit (Sigma–Aldrich). Briefly, the slides were incubated for 15 min in iron stain solution (prepared by mixing equal volumes of 4% w/v potassium ferrocyanide and 1.2 mM hydrochloric acid). Nuclear Fast Red was used as counterstain.
CD68 expression was analyzed by CD68 antibody staining (Dako; Ref-Nr.:M0876, Lot-Nr.: 00094,759; clone PG-M1) on the Ventana Benchmark Ultra (Roche) according to the manufacturer's instructions. Microscopy slides containing the tissue were preincubated with the EDTA-containing CC1 solution (Roche) for 16 min after which the slides were incubated with a 1:100 dilution of the CD68 antibody for 24 min.
The stained slides were digitally scanned with the Aperio AT2 (Leica).
4
Perls' Stain for Iron Detection
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Renal tissue slides from Sham- and CLP-treated mice were dewaxed in xylene and rehydrated in a series of alcohol solutions using decreasing concentrations. Perls’ stain was performed using the Iron Stain Kit (Sigma Aldrich, Germany) according to the manufacturer’s protocol. Slides were then washed in distilled water, counterstained with nuclear fast red solution (Sigma Aldrich), rapidly dehydrated, and mounted in entellan (Merck). Pictures were acquired using an Axioskop 40 (Zeiss, Wetzlar, Germany).
5
Histological Analysis of Spleen Tissue
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Following fixation in 10% formalin for 24 h, spleens were stored in 70% ethanol before further preparation. The tissue was embedded in paraffin and 7 μm cross-sections were cut with a microtome (Reichert-Jung, Germany). The sections were stained with hematoxylin and eosin. Slides were examined by light microscopy (Olympus, type CH2). Non-heme iron staining of spleen samples was analyzed using Iron Stain Kit (Sigma-Aldrich, HT20-1KT). Sections were prepared as described above. After mounting on glass slides, sections were deparaffinized, incubated with a working solution containing Perls' Prussian Blue for 30 min, counterstained with pararosaniline solution for 2 min, and analyzed under standard light microscopy (Olympus CH2).
6
Iron Deposition Detection Protocol
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Two sections per brain adjacent to the injection site were used for detecting iron deposition using an Iron Stain Kit (Sigma-Aldrich, St. Louis, MO, USA). Slides were incubated in a freshly prepared working iron stain solution for 15 min, washed in distilled water, and then counterstained with pararosaniline solution for 3 min. Data are presented as Prussian blue-positive area (mm2).
7
Perls' Prussian Blue Tissue Iron Staining
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We used Perls’ Prussian blue staining to determine the tissue iron content of each tumor sample. The staining was performed using the Iron Stain Kit (HT20-1KT, Sigma-Aldrich, St. Louis, MO, USA). Paraffin-embedded tissue samples were rehydrated, stained with Perls’ staining solution for 15 min and rinsed in running tap water for 5 min. The sections were then counterstained with nuclear fast red for 3 min and finally rinsed in running tap water for 5 min. Then, the sections were dehydrated in 70%, 96%, 100% ethanol and xylene and embedded using xylene-based mounting media.
8
Ferric Ammonium Sulfate Cytotoxicity Assay
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4T1 and 4T1-Qt cells were seeded at 2 × 105 cells/dish in 35 mm Petri dishes and cultured for 24 h. FAS (4 mM, 2 mM, 1 mM, 0.5 mM, 0.25 mM, and 0.13 mM) was added to the cells, and they were incubated with FAS for 24 h. Afterward, the cells were thoroughly washed with PBS, fixed by 4% formaldehyde in PBS, and then stained using Iron Stain Kit (Sigma-Aldrich), according to the manufacturer’s instructions. Images of 4T1 and 4T1-Qt cells were taken with an inverted microscope Primo Vert (Zeiss, Oberkochen, Germany).
9
Iron Deposition Detection Protocol
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Iron Stain Kit (Sigma-Aldrich, St. Louis, MO) was used to detect iron deposition. Slides were incubated in freshly prepared working iron stain solution for 15 minutes, washed in distilled water, and counterstained with pararosaniline solution for 3 minutes. Two sections per brain within the injection site were chosen for staining. Data presented as percentage of Prussian blue positive area in the hemisphere.
10
Quantitative tissue iron analysis
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