[Histonet] Re: Polyester wax

From:Stanislav Vitha

I have used the polyester wax (Steedman's wax) a lot for immunofluorescence 
in plants (with antibodies for actin, tubulin, FtsZ, GFP, c-myc, and 
other). I remember there were some old papers where the wax was used for 
decalcified bones.  Unfortunately these reprints were left behind during my 
last move, but a quick search found a couple papers that may be relevant:

1.      Sage, M., Polyethylene-Glycol Distearate 600 with 10 Percent 
1-Hexadecanol - Superior Embedding Wax for Warm Climates. Stain Technology, 
1972. 47(6): p. 313-&.
2.      Goodwin, J.R., Polyester Wax Processing of Tissues - Some New 
Techniques and Detailed Procedures. American Journal of Medical Technology, 
1974. 40(2): p. 45-49.
3.      Josephse.K, Technique for Isolating Enamel Organ of Rat Incisor for 
Histologic Studies. Scandinavian Journal of Dental Research, 1974. 82(3):=20
p. 229-238.
4.      Moores, B.D., Thin Histological Sections from Tissues Impregnated=20
with Digol Distearate. Medical Laboratory Sciences, 1976. 33(1): p. 73-77.
5.      Kusakabe, M., Sakakura, T., Nishizuka, Y., Sano, M., and Matsukage, 
A., Polyester Wax Embedding and Sectioning Technique for 
Immunohistochemistry. Stain Technology, 1984. 59(3): p. 127-132.
6.      Roholl, P.J.M., Dullens, H.F.J., Kleijne, J., Dubbink, E.J., and 
Denotter, W., Acid Ethanol Fixation and Polyester Wax Embedding Combines 
Preservation of Antigenic Determinants with Good Morphology and Enables 
Simultaneous Bromodeoxyuridine (Brdu) Labeling. Biotechnic & 
Histochemistry, 1991. 66(2): p. 55-62.
7.      Richardson, L.L. and Dym, M., Improved Adhesiveness of Polyester 
Wax Sections for Immunocytochemistry. Biotechniques, 1994. 17(5): p. 846-848.

As far as handling and sectioning, I was quite happy with the sectioning 
properties; I was cutting sections between 5 and 15 microns.
There are few tricks that are useful to make the sectioning work well; see 
the book chapter:

Vitha, S., Baluška, F., Jasik, J., Volkmann, D., and Barlow, P., Steedman's 
wax for F-actin visualization, in Actin: a Dynamic Framework for Multiple=20
Plant Cell Functions, C.J. Staiger, F. Baluška, D. Volkmann, and P. Barlow, 
Editors. 2000, Kluwer: Dordrecht, The Netherlands. p. 619-636.

You can download the PDF from 
http://www.izmb.de/volkmann/pdfs/  (Book-Steedman's_wax.pdf)

The disadvantage of the above method is that the ribbons sometimes do not=20
100%, or al least not all sections; for what I was doing it was not 
critical, I was more interested in cellular and subcellular immunostaining 
rather that looking at tissue and whole-organ morphology.

Also see my previous post to this forum (10-21-2004)

Stan Vitha

>Message: 11
>Date: Thu, 28 Oct 2004 00:33:36 -0700
>From: "Webster, Paul" 
>Subject: [Histonet] polyester wax
>         <4E8C1F1E4E8FA748B5487C50C91695F001280092@63-194-44-18.hei.org>
>Content-Type: text/plain;       charset="Windows-1252"
>Does anyone have experience with polyester wax as a substitute embedding=20
>medium? Is it possible to section decalcified bone and does it really 
>offer better results for immunohistology?
>Paul Webster.
>Paul Webster, Ph.D.
>House Ear Institute
>2100 W. 3rd St.
>Los Angeles
>CA 90057

Dr. Stanislav Vitha      vitha@mic.tamu.edu
Microscopy and Imaging Center
Texas A&M University
BSBW 119
College Station, TX 77843-2257

tel: 979-845-1129 (main desk)
tel: 979-845-1607 (direct link)
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