Difference between revisions of "Picrosirius Red Staining"
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| − | Picrosirius Red Staining<br /> | + | <font size="2"><font style="BACKGROUND-COLOR: #ccffcc"><strong><font size="4">Picrosirius Red Staining</font></strong><br /> |
| + | </font><br /> | ||
| + | <strong>To make the Picrosirius stain</strong><br /> | ||
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| − | |||
- 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 /> | - 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 /> | ||
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- 250 ml are needed for proper staining procedure.<br /> | - 250 ml are needed for proper staining procedure.<br /> | ||
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| − | + | <strong><br /> | |
| + | Staining :</strong> <br /> | ||
| + | <br /> | ||
- Paraffin embedded, 5 or 10 um thick sections are stained as follows:<br /> | - Paraffin embedded, 5 or 10 um thick sections are stained as follows:<br /> | ||
| + | <br /> | ||
Histoclear #1 - 3 minutes<br /> | Histoclear #1 - 3 minutes<br /> | ||
Histoclear #2 - 3 minutes<br /> | Histoclear #2 - 3 minutes<br /> | ||
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Histoclear #6 - 3 minutes<br /> | Histoclear #6 - 3 minutes<br /> | ||
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| − | Note : can add 0.2% phosphomolybdic acid in DD H2O for 2 minutes with quick rinse after 10 minute water step above.<br /> | + | <strong>Note : can add 0.2% phosphomolybdic acid in DD H2O for 2 minutes with quick rinse after 10 minute water step above.<br /> |
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| − | Note : In case of | + | Note : In case of freezing section,</strong> <br /> |
| + | 1) 60°C dry oven 30 min<br /> | ||
| + | 2) H2O rinse 10 min<br /> | ||
| + | 3) PS stain 90 min<br /> | ||
| + | 4) MQ washing </font> | ||
Latest revision as of 14:48, 21 November 2006
Picrosirius Red Staining
To make the Picrosirius stain
- 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.
- Add 0.1 gram sirius red F3BA per 100ml.
- Allow solution to stand for 24 hours.
- Stain should keep for a few months at room temperature, or 100 slides, whichever comes first.
- 250 ml are needed for proper staining procedure.
Staining :
- Paraffin embedded, 5 or 10 um thick sections are stained as follows:
Histoclear #1 - 3 minutes
Histoclear #2 - 3 minutes
Histoclear #3 - 3 minutes
100% EtOH - 3 minutes
95% EtOH - 3 minutes
70% EtOH - 3 minutes
H2O Rinse - 10 minutes
PS stain - 90 minutes
70% EtOH - 3 minutes
95% EtOH - 3 minutes
100% EtOH - 3 minutes
100% EtOH - 3 minutes
Histoclear #4 - 3 minutes
Histoclear #5 - 3 minutes
Histoclear #6 - 3 minutes
Note : can add 0.2% phosphomolybdic acid in DD H2O for 2 minutes with quick rinse after 10 minute water step above.
Note : In case of freezing section,
1) 60°C dry oven 30 min
2) H2O rinse 10 min
3) PS stain 90 min
4) MQ washing
