Histology sections were prepared from the knees of all rats and then stained with the Safranin O/Fast Green Staining Kit (Solarbio, Beijing, China). Pathologic improvements in the ankle joint were evaluated.
Safranin o fast green staining kit
Manufactured by Solarbio
Sourced in China
The Safranin O-fast green staining kit is a laboratory reagent used for staining biological samples. The kit contains the necessary solutions to perform a differential staining procedure, which highlights specific cellular structures or components within the sample.
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9 protocols using safranin o fast green staining kit
1
Histological Evaluation of Rat Knee
2
Paraffin-embedded IVD Tissue Staining
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After IVD tissues were embedded in paraffin, section (4 μm) was processed with a safranin O-fast green staining kit (Solarbio). For safranin O-fast green staining, histological images were acquired using light microscopy (Nikon, Tokyo, Japan), and the histological scores were determined according to previously described criteria21 (link). Image assessment was conducted by two independent observers.
3
Histological Assessment of Cartilage Repair
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At 8 weeks after repair, the rats were sacrificed. Rat knee joints were separated, washed with PBS and imaged. Samples were then fixed in 4% PFA solution for 24 h at 4 °C. All samples were decalcified by 20% ethylenediaminetetraacetic acid (EDTA) solution for 2 months, dehydrated, embedded in paraffin, and sectioned into 5 μm-thick sections. To evaluate the extent of tissue reconstruction, sections of all groups were stained with hematoxylin and eosin (H&E) staining solutions (Solarbio, China) and a safranin-O/fast green staining kit (Solarbio, China). Histological points of the repaired cartilage defect tissues were scored by 3 independent blinded observers using the International Cartilage Repair Society (ICRS) visual histological assessment [37 (link)].
4
Safranin O-Fast Green Staining Procedure
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The sample was fixed with paraformaldehyde, decalcified, dehydrated, and then embedded in paraffin. Then, the Safranin O-Fast Green Staining Kit (Solarbio, Beijing, China) was used for staining. Specifically, the sample was dewaxed in water, placed in freshly prepared Weigert's dye solution for 3-5 minutes, and then washed with water. Then, it was differentiated in acidic differentiation solution for 15 s, washed with distilled water for 10 min, dipped in the fast green staining solution for 5 min, and quickly washed with weak acid solution for 10-15 s to remove residual fast green. Then, it was placed in the Safranin O stain for 5 min, dehydrated in 95% ethanol and absolute ethanol, treated with xylene to make it transparent, and then sealed with optical resin for observations.
5
Histological Evaluation of Knee Osteoarthritis
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The knee joint samples were collected and fixed in 4% paraformaldehyde for 24 h. The bones were placed in 10% EDTA for 1 month to perform decalcification, embedded with paraffin, prepared as 5 μm thick sections, and stained with Safranin O-fast green staining kit (G1371, Solarbio, China) following manufacturer’s instruction. The images were captured and calculated. The scores of the femur and tibia were summed and presented as the OARSI score for each sample.
6
Histomorphological Assessment of Murine TMJ
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TMJ specimens were harvested from mice immediately after sacrifice and kept in 4% paraformaldehyde solution overnight at 4 °C. The decalcification, dehydration, and paraffin embedding of TMJ samples were performed following our previously established protocols.8 ,32 (link) The paraffin-embedded TMJ samples were cut into 5-micron sections and stained using a safranin O & fast green staining kit (Solarbio, Cat#G1371) as previously described.23 The SO&FG-stained sections were evaluated according to the OARSI scoring system in a double-blinded manner. The Safranin O-positive cartilage areas were determined by image J (version 1.53k) as previously described.47 (link)
7
Histochemical Analysis of Intervertebral Disc
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Hematoxylin-eosin (HE) staining was used to evaluate general histopathological findings of disc tissues. A modified Masson trichrome staining kit (G1345; Solarbio) was used to observe collagen fibers in the intervertebral disc. And the distribution and content of extracellular cartilage matrix in the intervertebral disc were stained by a modified Safranin O-fast green staining kit (G1371; Solarbio).
The expression and distribution of TGF-β1 and downstream p-Smad2 and p-Smad3 were observed by a standard immunohistochemical staining protocol. Sections were incubated with primary antibodies to TGF-β1 (1 : 200, ab92486; Abcam, Cambridge, UK) or p-Smad2 (1 : 200, AF3449; Affinity, Changzhou, China) or p-Smad3 (1 : 120, AF3362; Affinity) at 4°C overnight. IHC was performed using an UltraSensitive TM SP (rabbit) IHC Kit (KIT-9706; Maixin, Fuzhou, China). Triplicates of each sample were used for staining.
The expression and distribution of TGF-β1 and downstream p-Smad2 and p-Smad3 were observed by a standard immunohistochemical staining protocol. Sections were incubated with primary antibodies to TGF-β1 (1 : 200, ab92486; Abcam, Cambridge, UK) or p-Smad2 (1 : 200, AF3449; Affinity, Changzhou, China) or p-Smad3 (1 : 120, AF3362; Affinity) at 4°C overnight. IHC was performed using an UltraSensitive TM SP (rabbit) IHC Kit (KIT-9706; Maixin, Fuzhou, China). Triplicates of each sample were used for staining.
8
Femur Tissue Histological Analysis
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The femur tissue samples were fixed in 4% paraformaldehyde (PFA) for 24 h. The bones were placed in 10% EDTA for one month of decalcification, embedded with paraffin, prepared as 5-μm sections, and stained using a safranin O-fast green staining kit (G1371, Solarbio, Beijing, China) or an HE staining kit (G1120, Solarbio, Beijing, China) according to the manufacturer’s instructions. The images were captured under the CX43 microscope (Olympus Optical Co., Ltd., Tokyo, Japan) at ×200 magnification.
9
Histological Assessment of Rat Knee Joints
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Histological sections were prepared from the knees of all rats and then stained with the SafraninO-Fast Green Staining Kit (Solarbio, China). Pathological improvements in the ankle joint were then evaluated.
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