Sirius red fast green staining

Based on the identified signaling pathways, we assessed the specific ECM characteristics by Masson’s trichrome staining (Solarbio, China) and Sirius red/Fast Green staining (Solarbio, China) to identify the corresponding ECM characteristics of breast cancer tissues with distant metastasis. ECM collagen fibers were red-stained by Sirius red/Fast Green staining and blue-stained by Masson’s trichrome. The percentage of collagen area was estimated by ImageJ, and the organization of the collagen fibers was evaluated by fiber orientation distribution using the ImageJ plugin Orientation J.

In order to clarify the characteristics of the ECM corresponding to each ECM phenotype, we performed Masson’s trichrome staining (Solarbio, China) and Sirius red/Fast Green staining (Solarbio, China) to assess the ECM characteristics of breast cancer patients of different nodal status and pathological stages according to the manufacturer’s instructions. The density of collagen was estimated using red-stained collagen area of Sirius red/Fast Green staining and blue-stained collagen area of Masson’s trichrome using Image J in ten randomly selected microscopic fields per specimen. The degree of collagen fiber alignment was quantified using an image J plugin Orientation J.