Picro-Sirius Red Staining Kit 

Picro-Sirius Red Staining Kit, composed of hematoxylin and Sirius red staining solutions, can be used for staining collagen fibers. When used in immunohistochemical studies, Picro-Sirius Red Staining Kit can also visualize type I and type III collagen fibers.

Guidelines (The following are our recommended protocols. This protocol is for reference only and should be modified according to your specific needs.)
1. Fix tissue in 10% formalin fixative, and routinely dehydrate and embed.
2. Section 3-8 μm thick, dewax to water using xylene or Leagene environmentally friendly wax-removing and clearing solution.
3. Stain with hematoxylin solution for 3-10 min, then wash with water for 30-120 s.
4. Stain with Sirius red solution for 10-60 min, or 10-20 min for tissues that are easier to stain.
5. Rinse briefly with running water to remove staining solution from the surface of the sections.
6. Dehydrate with 95% ethanol for 1 min, then dehydrate twice with anhydrous ethanol for 1 min each time, clear twice with xylene or dewaxing and clearing solution for 1 min each time, and mount with neutral resin. Examine under a microscope.

Staining Results:
1. Ordinary Optical Microscope:
Collagen fibers appear red;
Muscle fibers and blood cells appear yellow;
Cell nuclei appear bluish-brown to black.

2. Polarized Light Microscope:
Type I collagen fibers appear orange-yellow or bright red;
Type III collagen fibers appear green.

Notes:
1. For clear visualization under a polarized light microscope, the section thickness should be 6-7 μm.
2. Without hematoxylin staining, the cell nuclei appear light red, providing better contrast between collagen and muscle fibers.
3. After Sirius red staining, prolonged treatment in low-concentration ethanol is not recommended, otherwise the yellow color of the muscle fibers will easily be removed.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

[Picro-Sirius Red Staining Kit]

Room temperature in continental US; may vary elsewhere.

Please store the product under the recommended conditions in the Certificate of Analysis.