Picrosirius staining

Frozen sections
Dry sections for 1 hour at RT.
Fixate for 20 min in 4% formaldehyde.
Wash 5 minutes in aqua dest.
Color 30 min in picrosirius-red.
Wash 3X 7 min in 0.2 N HCl.
Wash in aqua dest for 5 minutes.
Dehydrate in 70% EtOH quick, 2 x96% EtOH quick, 2 x 5 minutes 100% EtOH and 2 x 8 minutes
Ultraclear.
Mount in Entellian.
Paraffin sections

2X 8 minutes in Ultraclear.

2X 5 minutes in 100% EtOH.

2X 5 minutes in 96% EtOH.

5 minutes in 70% EtOH.

Wash 5 minutes in Aqua Dest.

Stain 30 minutes in Picrosirius-red

Wash 2X 5 minutes in 0.2 N HCl.

Wash in Aqua Dest for 5 minutes.

Dehydrate in 70% EtOH quick, 2 x96% EtOH quick, 2 x 5 minutes 100% EtOH and 2 x 8 minutes
Ultraclear.

Mount slides in Entellan.

Picrosirius Red (100 ml)

1.1 gram Sirius Red F3B
100 ml saturated picrine acid solution (pH=2.0) (commercial)
Dissolve and filtrate
0.2N HCL: Dilute 12N HCl (37%) 60 times in aqua dest.