Puchtler's Picro-Sirius red
for Collagen


Picro-sirius red
Sirius red F3B 0.5 g
Saturated aqueous picric acid 500 mL
Acetic acid water
Acetic acid, glacial 5 mL
Distilled water 11 mL

Tissue sample
5µ paraffin sections of neutral buffered formalin fixed tissue are suitable.


  1. Bring sections to water.
  2. Optionally, stain nuclei with an acid resistant nuclear stain.
  3. Wash well with water.
  4. Place into Picro-sirius red solution for 1 hour.
  5. Wash with two changes of acetic acid water.
  6. Dehydrate with ethanol, clear with xylene and mount with a resinous medium.

Expected results
Light microscopy

Polarising microscopy


  1. This method can be used as an alternative for van Gieson's stain or, using polarising microscopy, as a sensitive method for collagen.


Kiernan. J.A., (1999)
Histological and histochemical methods: Theory and practice, Ed. 3
Butterworth Heinemann, Oxford, UK.




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