Re: IMAGING OF CELLULOSE

Posted by Rosemary.White on
URL: http://confocal-microscopy-list.275.s1.nabble.com/IMAGING-OF-CELLULOSE-tp7582017p7582035.html

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Acridine orange - the "RNA stain" also stains cell walls very nicely and
uniformly, especially cotton cell walls. It overcomes the irritating
effect of laser polarisation seen when using calcofluor white.

Propidium iodide will stain the pectic components of cotton fibres, but
you also see the effect of laser polarisation there too.

On 8/05/14 6:00 AM, "Lloyd Donaldson" <[hidden email]>
wrote:

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>Cotton does contain small amounts of protein. It is unlikely that the
>cellulose would fluoresce.
>
>-----Original Message-----
>From: Confocal Microscopy List [mailto:[hidden email]]
>On Behalf Of Guy Cox
>Sent: Thursday, 8 May 2014 3:54 a.m.
>To: [hidden email]
>Subject: Re: IMAGING OF CELLULOSE
>
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>I totally endorse TDE as an embedding medium but I am puzzled by acrolein
>inducing fluorescence in cellulose.  Can someone explain the chemistry?
>I thought acroleon reacted with proteins.
>
>                                                            Guy
>________________________________________
>From: Confocal Microscopy List [[hidden email]] on
>behalf of Stanislav Vitha [[hidden email]]
>Sent: 08 May 2014 00:04
>To: [hidden email]
>Subject: Re: IMAGING OF CELLULOSE
>
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>I have recently done some confocal imaging of cotton fibers. For deeper
>penetration, less scattering in single photon fluorescence imaging,
>rather than Calcofluor White I suggest staining with orange or
>red-fluorescing dyes - some have been already mentioned, like Congo Red.
>The fluorescent version of Periodic Acid-Schiff staining works great (use
>propidium iodide as pseudo-Schiff reagent; Truernit, E., H. Bauby, et al.
>(2008). "High- resolution whole-mount imaging of three-dimensional tissue
>organization and gene expression enables the study of Phloem development
>and structure in Arabidopsis." Plant Cell 20(6): 1494-1503).
>As an alternative to fluorescence staining, I have also used vapor
>fixation with acrolein, which tends to induce very strong
>autofluorescence, at least in cotton fibers.  This is useful if you do
>not want to expose the fibers to water and induce swelling.
>For imaging with oil immersion objectives I used 2,2-thioidiethanol as a
>mounting medium with good success.
>Staudt, T., M. C. Lang, et al. (2007). "2,2'-thiodiethanol: a new water
>soluble mounting medium for high resolution optical microscopy."
>Microscopy Research and Technique 70(1): 1-9.
>
>Stan Vitha
>Microscopy and Imaging Center
>Texas A&M University
>
>
>
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