SPI-Chem™ Lead Citrate, Trihydrate
Additional information
Another reference for the use of lead citrate is the following:
Venable, J. M. and Coggeshall, R., J. Cell Biol. 25, 407-408 (1965).
This protocol calls for the following:
1. Add 0.01-0.04 gms lead citrate
and 0.1 ml of 10N sodium hydroxide to
10 ml distilled water to some type of screw-topped vial. Use distilled
water that has been freshly boiled for at least 5 minutes to insure solutions
are carbon dioxide free. This step is very important.
2. Sonicate the mixture and/or vigorously
shake until all of the lead citrate is completely dissolved. You will know
when it is all dissolved by the solution turning clear.
3. Some workers recommend centrifuging before use. Others allow the solution
to "stand" 30 minutes without agitating the bottle.
4. Yet others say "use stain immediately", since deterioration of the stain's
efficacy starts immediately. In any case, it is good practice not to use any of
the staining solution from the bottom of the vial.
5. Usual staining time is 1-5 minutes.
Additional comments (culled from the Microscopy Listsever) on this subject:
- This formula is thought to be much more intense than the formula originally
published in Reynolds, J. Cell Bio. 17, 208 (1963).
- We can not over emphasize the high sensitivity of this stain to carbon
dioxide.
- To avoid precipitation, make fresh staining solution each time.
- Keep staining times short and always rinse in carbon dioxide-free
distilled water.
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Sunday March 21, 2010
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