Trichrome staining was performed with Trichrome Stain (Masson) Kit (Abcam, Cambridge, MA USA). The manufacturer's instructions were used. Briefly, the cryosections were thawed and Bouin's solution was used to fix the samples. Weigert's Iron Hematoxylin was used for nuclear staining, Biebrich Scarlet/Acid Fuchsin Solution for cytoplasmic staining and aniline blue for collagen lattice. The samples were observed under an inverted Olympus microscope (Tokyo, Japan). The MIPAR Image Analysis Software (7 (link)) recipe used to quantify the images utilizes stain deconvolution approach as described elsewhere (8 (link)). For our analysis, the recipe separated the trichrome stain into grayscale representations for effective contrast. The grayscale images are then segmented to identify total tissue, excluding slide background (white space), and to isolate collagenous tissue. The collagen area fraction measurement is made relative to the total tissue area. Each group comprised of four different animals.
Trichrome stain masson kit
Manufactured by Abcam
Sourced in United States
The Trichrome Stain (Masson) Kit is a set of reagents used for the histological staining of tissue sections. The kit provides the necessary components to perform a trichrome staining technique, which is commonly used to differentiate various tissue types, such as collagen, muscle, and connective tissue.
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2 protocols using trichrome stain masson kit
1
Quantifying Collagen in Tissue Sections
2
Histological Analysis of Liver and EWAT
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Liver and EWAT were embedded in paraffin after being fixed overnight in 10% Neutral buffered formalin. Then, xylene was used to deparaffinize portions that were five micrometers thick, and a graded ethanol series was used to dehydrate them. Hematoxylin (2 g/L) was used to stain the tissue slices for 15 minutes, and eosin (0.1 percent in 0.0003 percent acetic acid) was used for 10 minutes at room temperature. Masson stain used by Trichrome Stain (Masson) Kit (abcam, Cambridge, USA). For immunohistochemistry, endogenous peroxidase activity was quenched with 3% H 2 O 2 in water for 10 min. Non-specific staining was minimized with 5% normal goat serum for 50 min. Next, sections were subject to antigen retrieval, as needed, and incubated with primary and second antibodies for 2 hours at room temperature. The antibodies used in this study are shown in Table 1. Images were obtained using Olympus IX71 microscope.
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