For histological analysis, the osteochondral (OC) grafts were fixed in a 4% buffered formaldehyde solution (VWR, Radnor, PA, USA) for up to 1 week. Decalcification was then performed under constant agitation using the Osteosoft solution (Merck, Burlington, MA, USA) for 4 to 6 weeks. Following decalcification, the OC grafts were embedded in Tissue-Tek® OCT (Optimal Cutting Temperature, VWR, Radnor, PA, USA) and stored at −80 °C. Sectioning was performed using the CryoStarTM NX70 Cryostat (Thermo Fischer Scientific, Waltham, MA, USA) with a knife temperature of −25 °C and a chamber temperature of −20 °C. Then, 6 µm sections were obtained and subjected to Safranin O/Light Green staining. Images were captured using a Leica DM-1000 microscope and processed using Leica IM500 Image Manager software Version 5 (Leica, Wetzlar, Germany).
Buffered formaldehyde solution
Manufactured by Avantor
Sourced in United States
4% buffered formaldehyde solution is a fixative used in various laboratory applications. It is a water-based solution that contains 4% formaldehyde and a buffer to maintain a stable pH. This product is commonly used for the preservation and fixation of biological samples, such as tissues or cells, prior to further analysis or processing.
Automatically generated - may contain errors
4 protocols using buffered formaldehyde solution
1
Histological Analysis of Osteochondral Grafts
2
Histological Analysis of Osteochondral Plugs
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For histological analysis, osteochondral plugs were submerged in 4% buffered formaldehyde solution (VWR, Radnor, PA) for ≤7 days, and decalcified for 4–6 weeks under constant agitation using the Osteosoft solution (Merck, Burlington, MA). After decalcification, the samples were embedded in Tissue‐Tek O.C.T. and stored at −80°C. Cryosectioning was performed using the CryoStar NX70 Cryostat (Thermo Fischer Scientific, Waltham, MA) to obtain 6‐µm sections. Subsequently, the samples were prepared for Safranin O staining and Fast‐green counterstaining. Histological images were captured using a Leica microscope DM‐1000 and processed using the Leica Manager software (Leica, Wetzlar, Germany).
3
Histological Analysis of Osteochondral Grafts
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For histological analysis, osteochondral grafts were fixed in 4% buffered formaldehyde solution (VWR, Radnor, PA) for up to 1 week and decalcified under constant agitation using Osteosoft solution (Merck, Burlington, MA). After decalcification (duration of 4–6 weeks), the osteochondral grafts were embedded in Tissue‐Tek® OCT (Optimal Cutting Temperature, VWR, Radnor, PA) and stored at −80°C. Sectioning was done using the CryoStar™ NX70 Cryostat (Thermo Fischer Scientific, Waltham, MA), with −25°C for the knife temperature and −20°C for the chamber temperature. A 6 μm sections were obtained and processed for Safranin O staining. Images were taken with a Leica microscope DM‐1000 and processed using the Leica Manager software (Leica, Wetzlar, Germany).
4
Histological Analysis of Osteochondral Grafts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For histological analysis, osteochondral grafts were fixed in 4% buffered
formaldehyde solution (VWR, Radnor, PA, USA) for up to 1 week and decalcified
under constant agitation using Osteosoft solution (Merck, Burlington, MA, USA).
After decalcification (duration of 4-6 weeks), the osteochondral grafts were
embedded in Tissue-Tek OCT (optimal cutting temperature, VWR, Radnor, PA, USA)
and stored at −80°C. Sectioning was done using the CryoStar NX70 Cryostat
(Thermo Fischer Scientific, Waltham, MA, USA), with −25°C for the knife
temperature and −20°C for the chamber temperature. Six-micrometer sections were
obtained and processed for safranin O/light green staining. Images were taken
with a Leica DM-1000 microscope and processed using the Leica Manager software
(Leica, Wetzlar, Germany). To quantify changes within the cartilage sections
stained with safranin O/light green, a modified Mankin scoring system was used
(Table 3
The assessment was done by 5 independent observers with a maximum score
of 15.
formaldehyde solution (VWR, Radnor, PA, USA) for up to 1 week and decalcified
under constant agitation using Osteosoft solution (Merck, Burlington, MA, USA).
After decalcification (duration of 4-6 weeks), the osteochondral grafts were
embedded in Tissue-Tek OCT (optimal cutting temperature, VWR, Radnor, PA, USA)
and stored at −80°C. Sectioning was done using the CryoStar NX70 Cryostat
(Thermo Fischer Scientific, Waltham, MA, USA), with −25°C for the knife
temperature and −20°C for the chamber temperature. Six-micrometer sections were
obtained and processed for safranin O/light green staining. Images were taken
with a Leica DM-1000 microscope and processed using the Leica Manager software
(Leica, Wetzlar, Germany). To quantify changes within the cartilage sections
stained with safranin O/light green, a modified Mankin scoring system was used
(
The assessment was done by 5 independent observers with a maximum score
of 15.
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