[Taxacom] Selective staining of suberified cells in Poaceae

[Lewis, Deborah A [EEOBS]]

Martin,

I asked Harry (Jack) Horner, who directs our microscopy and nano-imaging facility, for his suggestions. Here is his reply:
With regard to suberin staining, this is a challenge as most 'specific' stains do stain other lipid-containing substrates.  The sudan dyes stain suberin but also other substrates.

I have two alternate suggestions:
-Use sudan blue after treating sections with concentrated sulfuric acid (to remove other lipophilic substrates), washing with distilled water and then staining.  Latter treatment also may be tried with sudan IV and sudan black B.

-Also try the sulfuric acid treatment and wash; and then add 0.02 M phosphate buffer, pH 9.1, coverslip and view with UV microscope.  Suberin fluoresces yellow.

Hope this will help!
Deb Lewis

Deborah Q. Lewis, Curator
Ada Hayden Herbarium                        Phone:  [1] 515-294-9499
Iowa State University                            Email:    dlewis at iastate.edu<mailto:dlewis at iastate.edu>
340 Bessey                                                 FAX:       [1] 515-294-1337
Ames, IA  50011-1020                            Web:      www.herbarium.iastate.edu<http://www.herbarium.iastate.edu>

-----Original Message-----
From: taxacom-bounces at mailman.nhm.ku.edu [mailto:taxacom-bounces at mailman.nhm.ku.edu] On Behalf Of Martin Dub?
Sent: Monday, May 13, 2013 12:43 PM
To: taxacom at mailman.nhm.ku.edu
Subject: [Taxacom] Selective staining of suberified cells in Poaceae

For those anatomicaly oriented agrostologists,

Stains that color suberin (in so-called suberous short cells for ex.) also color other parts of tissue (chlorenchyma for ex.) making such preparations often difficult to observe, unless these latter are removed.  However, in certain cases (as with lemmas), removing the unwanted tissue is hardly possible.   I would greatly appreciate receiving suggestions from experienced botanists about a more selective stain for the suberin containing cells, i.e. not coloring cuticle or cell walls or cytoplasm.  P.S.: I have been using lactophenol cotton blue which is not selective enough.

Thanks.

Martin Dub?, Ph.D.
Associate professor

Campus d'Edmundston
Universit? de Moncton
Edmundston, NB
CANADA  E3V 2S8


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