Metaphase slides were washed for 10 min with phosphate buffer before chromomycin A3 (CMA3) solution (10 mg/ml) addition (Sigma) and left for 2 hours at RT in a dark, humified chamber. The slides were then rinsed in NaCl-HEPES (4-(2-Hydroxyethyl)piperazine-1-ethanesulfonic acid) buffer and stained for 15 min in methyl green solution (Sigma). After two washes in NaCl-HEPES buffer, the antifading (Vectashield H-1200/isopropilgallate—1:300; Vector Laboratories, Burlingame, CA) was added. The slides were stored at 4 °C for 3 days in the dark, prior observation on a fluorescent Zeiss Axioplan microscope with CCD.
Methyl green solution
Manufactured by Merck Group
Sourced in United States
Methyl green solution is a laboratory reagent used for various staining and analytical purposes. It is a stable and concentrated solution of the dye methyl green, which is commonly employed in biological and histological applications. The core function of this product is to provide a consistent and reliable source of the methyl green dye for use in research and diagnostic procedures.
Automatically generated - may contain errors
Lab products found in correlation
Axioplan, by Zeiss (1 mentions)
3 3 diaminobenzidine (dab), by Vector Laboratories (1 mentions)
Bx51 dot slide, by Olympus (1 mentions)
S7100, by Merck Group (1 mentions)
Trap staining solution, by Merck Group (1 mentions)
H e staining, by Solarbio (1 mentions)
Ab4073, by Merck Group (1 mentions)
Eclipse te2000, by Nikon (1 mentions)
Vectashield h 1000, by Vector Laboratories (1 mentions)
Chromomycin a3, by Merck Group (1 mentions)
6 protocols using methyl green solution
1
Chromomycin A3 Staining of Metaphase Slides
2
Femur Alkaline Phosphatase Staining
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The femur paraffin sections were dewaxed and stained using an ALP Staining Kit (Sangon Biotech, C520019-0005, Shanghai, China) according to the manufacturer’s instructions. Briefly, the sections were stained with ALP solution at 37 °C for 30 min and then counterstained with 2% methyl green solution (Sigma, M8020, MO, USA) for 10 min at room temperature.
3
TUNEL Assay for Apoptosis Detection
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A terminal deoxynucleotidyltransferase-mediated dUTP nick-end labeling (TUNEL) assay was performed according to the manufacturer’s instructions (Cat.No.#S7100, Millipore). Heart tissues were fixed in 4% formaldehyde and embedded in paraffin. Tissue sections (5 μm thickness) were deparaffinized, dehydrated, and rinsed with PBS. The slides were treated with 3% hydrogen peroxide and TdT enzyme at room temperature for one hour followed by digoxygenin-conjugated nucleotide substrate at 37 °C for 30 min. Nuclei were stained with DAB (Vector Laboratories, Burlingame, CA, USA) for five minutes, and the slides were counterstained with 0.5% methyl green solution (Sigma Aldrich, St. Louis, MO, USA). The slides were observed by a virtual microscopy (BX51/dot Slide; Olympus, Tokyo, Japan).
4
Mouse Femur Histological Analysis
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The mouse femurs were fixed and decalcified, and then embedded in paraffin. Sections of 4.5 μm were prepared. H&E staining (Solarbio, Beijing, China) was performed as the manufacturer’s instruction. The stained sections were observed using an inverted microscope (IX81, Olympus).
TRAP staining was performed according to the procedure described previously (Liu et al., 2016 (link)). TRAP staining solution (Sigma) was added to cover the tissue sections. After incubating at 37°C for 40 min, the sections were stained with 0.1% methyl green solution (Sigma) for 10 s. The stained sections were observed using an inverted microscope (IX81, Olympus).
TRAP staining was performed according to the procedure described previously (Liu et al., 2016 (link)). TRAP staining solution (Sigma) was added to cover the tissue sections. After incubating at 37°C for 40 min, the sections were stained with 0.1% methyl green solution (Sigma) for 10 s. The stained sections were observed using an inverted microscope (IX81, Olympus).
5
Histopathological Analysis of Liver Tissue
6
Karyotyping hTERT RPE-1 Cell Line
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To confirm the chromosomal structure of the assembly, a karyotype for hTERT RPE-1 cells was generated. Metaphase spreads from hTERT RPE-1 cells were washed with a phosphate buffer (0.07 M NaH2PO4, 0.07 M Na2HPO4, 1 mM NaCl, pH 6.8) followed by 2-hour incubation with Chromomycin A3 (0.6 mg/ml, Sigma-Aldrich C2659) at RT in a dark, moist chamber. Slides were then washed with NaCl-HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) (0.15 M NaCl, 5 mM HEPES), stained for 15 minutes in a methyl green solution (0.1 mM, Sigma-Aldrich) and washed twice with NaCl-HEPES. The antifading (Vectashield H-1000; Vector Laboratories) isopropilgallate 1:300 was added to the slides before storing them at 4°C for 3 days in a dark, moist chamber. Images were acquired using a Thunder fluorescent widefield microscope (Leica) at a 100X magnification.
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