Leigh

There is no really specific stain for lignin for either fluorescence or brightfield. For fluorescence we find autofluorescence to be the most reliable. Stains often give varying responses depending on whether the material is embedded or not, and what its embedded in.
However if you are using unembedded material then phloroglucinol will be good if it is a gymnosperm. I see you are working on Plantago so the Maule reagent is likely to be better.
For embedding try LR White hard formulation or SPurr resin. Spurr resin can be removed after sectioning which might improve staining. Try silane coated slides if you have adhesion problems.


Dr Lloyd Donaldson
Senior Scientist
Scion - Next Generation Biomaterials
49 Sala St. Rotorua
Private Bag 3020, Rotorua 3046
NEW ZEALAND
Ph: 64 7 343 5581
Fx: 64 7 343 5507
www.scionresearch.com



-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of Dale Callaham
Sent: Saturday, 23 January 2010 12:43 a.m.
To: [log in to unmask]
Subject: Re: off topic: lignin staining

Leigh,

This is just a suggestion - I don't work with HEMA embedments, but I
have used the stain with material in LRWhite. The pararosanaline-based
Schiff's Reagent will stain the free amines of lignin and there are
typically few interferences in "acetic-alcohol" fixed plant tissues. It
is both colored and highly fluorescent with a Rhodamine exciter set. The
fluoresence is very stable and resistant to bleaching.

See the image on our web page - secondary wall thickenings of Plantago
xylem.
http://www.bio.umass.edu/microscopy.

Dale


Leigh Silvester wrote:
> Hi
>
> A colleague is trying to stain some wood material for lignin using
> phloroglucinol-HCl.
>
> Due to the nature of the material it is currently being set in HEMA -
> hydroxyethylmethacrylate.
>
> Problem is that the stain does not appear to be compatible with this
> resin.
> The stain works fine on lose preparations. He is also aware of the light
> sensitivity issue.
>
> Any one suggest any pretreatments for the HEMA sections?
> Perhaps there is an alternative lignin stain that is compatible with
> this resin?
>
> We have tried other resins but so far the HEMA one gives the best
> sections with this material.
>
> Any suggestions are very welcome.
>
> Regards
>
> Leigh Silvester
> ==================================================================
> Division of Animal Sciences
> University of Nottingham
> Sutton Bonington Campus
> Loughborough
> Leicestershire
> LE12 5RD
> T: +44 (0)115 9516045
> F: +44 (0)115 9516302
> E: [log in to unmask]
>
> NOTE - NEW TELEPHONE NUMBER
> ==================================================================
> Do you need to print this email?
> ==================================================================
>
> This message has been checked for viruses but the contents of an attachment
> may still contain software viruses which could damage your computer system:
> you are advised to perform your own checks. Email communications with the
> University of Nottingham may be monitored as permitted by UK legislation.

Disclaimer: This e-mail and any attachments may contain information which is confidential or subject to copyright. If you receive this e-mail in error, please delete it.
Scion does not accept responsibility for anything in this e-mail which is not provided in the  course of Scion's usual business or for any computer virus, data corruption, interference or delay arising from this e-mail.