Posted by
Lloyd Donaldson on
URL: http://confocal-microscopy-list.275.s1.nabble.com/IMAGING-OF-CELLULOSE-tp7582017p7582034.html
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Cotton does contain small amounts of protein. It is unlikely that the cellulose would fluoresce.
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From: Confocal Microscopy List [mailto:
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Sent: Thursday, 8 May 2014 3:54 a.m.
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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.
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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
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