Posted by
Guy Cox-2 on
URL: http://confocal-microscopy-list.275.s1.nabble.com/IMAGING-OF-CELLULOSE-tp7582017p7582033.html
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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
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From: Confocal Microscopy List [
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Sent: 08 May 2014 00:04
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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