Hi. I am a novice to histology as well as the analysis of histological images and I have used Sirius Red staining to stain for collagen in my lung samples. Can anyone please help me with a protocol or SOP with Image J (or any other tool) which I can use to quantify the area of my slide covered by fibrotic tissue (often accompanied by excessive and disordered collagen depostiion)? I have attached one of my images below in which I have tried to identify what I think are areas of fibrosis and areas of normal collagen cocentration. I want to quantify areas of fibrosis and I also want to avoid quantifying areas with normally high collagen content such as areas of the large airways.
Sirius red will stain collagen in red. I am also open to proposals of other tools that can help me too.
Thank you in advance for your kind help.