Puchtler's Picro-Sirius red
for Collagen
Materials
Picro-sirius red
| Material | Amount | |
|---|---|---|
| Sirius red F3B | 0.5 | g |
| Saturated aqueous picric acid | 500 | mL |
Acetic acid water
| Material | Amount | |
|---|---|---|
| Acetic acid, glacial | 5 | mL |
| Distilled water | 11 | mL |
Tissue Sample
5µ paraffin sections of neutral buffered formalin fixed tissue are suitable.
Protocol
- Bring sections to water.
- Optionally, stain nuclei with an acid resistant nuclear stain.
- Wash well with water.
- Place into Picro-sirius red solution for 1 hour.
- Wash with two changes of acetic acid water.
- Dehydrate with ethanol, clear with xylene and mount with a resinous medium.
Expected Results
Light Microscopy
- Nuclei – black if stained
- Collagen – red
- Cytoplasm – yellow
Polarising Microscopy
- Large fibres – yellow or orange birefringence
- Thin fibres – green birefringence
Notes
- This method can be used as an alternative for van Gieson’s stain or, using polarising microscopy, as a sensitive method for collagen.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Kiernan. J.A., (1999)
Histological and histochemical methods: Theory and practice, Ed. 3
Butterworth Heinemann, Oxford, UK.
