For xylem cell staining, haustoria were excised from roots, immersed in 10% (w/v) KOH (Wako Chemicals, Osaka, Japan) for 15 min at 90°C, rinsed three times with water, immersed in 0.1% (w/v) Safranin O (Wako Chemicals, Osaka, Japan) solution for 5 min at 90°C, and rinsed three times with water. The stained haustoria were cleared by soaking samples in clearing solution [chloral hydrate (2.5 g ml−1) (Nacalai Tesque), 33% (v/v) glycerol] from 3 hours to overnight before observation. Safranin O–stained xylem cells were observed with a light microscope (Leica DMI3000 B) and an LSM700 laser-scanning confocal microscope (Carl Zeiss) with excitation and emission wavelength suitable for fluorescent images of red fluorescent protein (RFP).
Safranin o
Manufactured by Leica
Sourced in Belgium
Safranin O is a red dye commonly used in biological and medical laboratory applications. It is a water-soluble stain primarily used for staining nucleic acids, such as DNA and RNA, as well as for staining plant tissues. Safranin O is a versatile dye that can be applied in various staining techniques to provide visual contrast and enhance the identification of cellular structures under a microscope.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 700, by Zeiss (2 mentions)
Safranin o, by Merck Group (2 mentions)
Chloral hydrate, by Nacalai Tesque (1 mentions)
Safranin o, by Fujifilm (1 mentions)
Dmi3000b, by Leica (1 mentions)
Hematoxylin, by Leica (1 mentions)
Fast green, by Merck Group (1 mentions)
Rm2145, by Leica (1 mentions)
H3 filter cube, by Leica (1 mentions)
Hematoxylin eosin, by Leica (1 mentions)
Hematoxylin, by Merck Group (1 mentions)
2 protocols using safranin o
1
Xylem Cell Staining Protocol
2
Histological Analysis of Tissue-Engineered Constructs
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At the end of 28 days of culture, beads were fixed with 4% paraformaldehyde in 100 mM sodium-cacodylate buffer, pH 7.4, for 4 h at 4°C; 20 mM CaCl2 was added to prevent disintegration of the beads. After washing overnight at 4°C in 100 mM sodium-cacodylate buffer, the beads were dehydrated in graded series of methanol, placed in a xylene wash before being embedded in paraffin. Five μm sections were cut with a microtome (Leica RM 2145; Leica, Diegem, Belgium) rehydrated and stained with hematoxylin-eosin, alcian blue or safranin-O/fast green. To visualize chitosan-FITC, hematoxylin-eosin stained sections of beads were observed using H3 filtercube (Leica). Alcian blue and safranin-O/fast green staining methods were used to analyze cartilage tissue formation. For safranin-O/fast green staining, sections were counterstained with hematoxylin (Merck, Darmstadt, Germany) and fast green (Merck) to visualize cells and cell nuclei, respectively. safranin-O (Sigma) was used for visualization of glycosaminoglycans (GAGs) in red. Sections were also stained with alcian blue solution (1% in acetic acid) to visualize extracellular GAGs in blue.
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