0
edits
Changes
From Opengenome.net
no edit summary
Picrosirius Red Staining<br />
<br />
To make the Picrosirius stain<br />
- Make saturated aqueous picric acid : heat 2~3 heaping teaspoons picric acid in 500+ ml distilled water, then cool. We use a magnetic stirrer / hot plate. If yellow needle-like crystals are not present when cool, then temperature was not high enough, or more acid is needed. Try heating again. Note : dry picric acid not in solution is very dangerous (combustible), so keep wet! Drain liquid for stain ; crystals may be used again.<br />
<br />
- Add 0.1 gram sirius red F3BA per 100ml.<br />
- Allow solution to stand for 24 hours.<br />
- Stain should keep for a few months at room temperature, or 100 slides, whichever comes first.<br />
- 250 ml are needed for proper staining procedure.<br />
<br />
Stainig : <br />
- Paraffin embedded, 5 or 10 um thick sections are stained as follows:<br />
Histoclear #1 - 3 minutes<br />
Histoclear #2 - 3 minutes<br />
Histoclear #3 - 3 minutes<br />
100% EtOH - 3 minutes<br />
95% EtOH - 3 minutes<br />
70% EtOH - 3 minutes<br />
H2O Rinse - 10 minutes<br />
PS stain - 90 minutes<br />
70% EtOH - 3 minutes<br />
95% EtOH - 3 minutes<br />
100% EtOH - 3 minutes<br />
100% EtOH - 3 minutes<br />
Histoclear #4 - 3 minutes<br />
Histoclear #5 - 3 minutes<br />
Histoclear #6 - 3 minutes<br />
<br />
Note : can add 0.2% phosphomolybdic acid in DD H2O for 2 minutes with quick rinse after 10 minute water step above.<br />
<br />
Note : In case of
<br />
To make the Picrosirius stain<br />
- Make saturated aqueous picric acid : heat 2~3 heaping teaspoons picric acid in 500+ ml distilled water, then cool. We use a magnetic stirrer / hot plate. If yellow needle-like crystals are not present when cool, then temperature was not high enough, or more acid is needed. Try heating again. Note : dry picric acid not in solution is very dangerous (combustible), so keep wet! Drain liquid for stain ; crystals may be used again.<br />
<br />
- Add 0.1 gram sirius red F3BA per 100ml.<br />
- Allow solution to stand for 24 hours.<br />
- Stain should keep for a few months at room temperature, or 100 slides, whichever comes first.<br />
- 250 ml are needed for proper staining procedure.<br />
<br />
Stainig : <br />
- Paraffin embedded, 5 or 10 um thick sections are stained as follows:<br />
Histoclear #1 - 3 minutes<br />
Histoclear #2 - 3 minutes<br />
Histoclear #3 - 3 minutes<br />
100% EtOH - 3 minutes<br />
95% EtOH - 3 minutes<br />
70% EtOH - 3 minutes<br />
H2O Rinse - 10 minutes<br />
PS stain - 90 minutes<br />
70% EtOH - 3 minutes<br />
95% EtOH - 3 minutes<br />
100% EtOH - 3 minutes<br />
100% EtOH - 3 minutes<br />
Histoclear #4 - 3 minutes<br />
Histoclear #5 - 3 minutes<br />
Histoclear #6 - 3 minutes<br />
<br />
Note : can add 0.2% phosphomolybdic acid in DD H2O for 2 minutes with quick rinse after 10 minute water step above.<br />
<br />
Note : In case of
