Re: fixative question for chemistry experts

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From:Geoff McAuliffe <mcauliff@UMDNJ.EDU>
To:Marcia Bentz <mb7x@virginia.edu>
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Date:Fri, 03 Sep 1999 17:20:50 -0400
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Marcia Bentz wrote:

> Hi all,
> Just wondering if anyone out there's heard of a fixative called PLP.

Marcia:

    PLP, periodate lysine paraformaldehyde was invented by McLean and Nakane in
1974 (J. Histochem Cytochem 22:1077). It is useful for immunostaining and even
gives decent ultrastructure without glutaraldehyde. It is not the same as
"routine" formaldehyde mixtures (which contain neither lysine nor periodate).
    I make it as follows:

1. Prepare 50 ml of 8% paraformaldehyde as usual.
2. Mix with 50 ml of 0.2M phosphate buffer. Subsequent ingredients will lower
the pH so use a phosphate buffer that is more alkaline than usual.
3. Add 1.37 g lysine, mix.
4. Just before use add 0.214 g sodium meta-periodate and mix.
5. Adjust the pH to 7.2 to 7.4.

Geoff
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Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
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mcauliff@umdnj.edu
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