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Yes, exactly as Lloyd says below.
There are two other things to overcome here - light scattering with depth
and light absorption with depth, and I think these are touched on in some
of the literature Lloyd refers to.
The inability to penetrate deeply - a depth/scattering problem - can be
ameliorated by matching the refractive index of the immersion medium with
that of the fabric. This might need to be determined empirically, but will
be around 1.5. Glycerol, at RI 1.47, is close.
However, there is also a tendency to overstain cellulose - it looks great
when it's really bright, but if it's very thoroughly stained, the fibres
will absorb more of the laser light, leaving less to penetrate into deeper
layers. You may find that with lighter staining, you can resolve some of
the deeper layers of the material. If your material has been dyed with
something very absorbant, this will be difficult....
cheers,
Rosemary White
Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia
T 61 2 6246 5475
F 61 2 6246 5334
E [log in to unmask]
On 7/05/14 5:55 AM, "Lloyd Donaldson" <[log in to unmask]>
wrote:
>For confocal microscopy use congo red, pontamine fast scarlet or
>calcofluor to stain cellulose. Alternatively fluorescently labelled
>cellulose binding domains are available or you can just fluorescently
>label some cellulase enzyme. However visualising beyond the surface layer
>will be difficult. To see the cross-section you can embed the paper in
>resin and cut cross-sections and stain them with toluidine blue for
>brightfield microscopy. You can also polish the cross-section of embedded
>paper, stain with iodine vapour and image with SEM in backscattered mode
>(also useful for visualising clay fillers or coatings. There is an
>extensive literature on this topic mainly in specialist pulp & paper
>journals.
>
>
>Dr Lloyd Donaldson
>Project Leader Microscopy & Wood Identification
>Senior Scientist Plant Cell Walls & Biomaterials
>Scion Forests, Products, Innovation
>49 Sala Street, Rotorua 3010
>New Zealand
>Ph 07 343 5581
>www.scionresearch.com
>
>
>
>-----Original Message-----
>From: Confocal Microscopy List [mailto:[log in to unmask]]
>On Behalf Of Philip Oshel
>Sent: Wednesday, 7 May 2014 2:48 a.m.
>To: [log in to unmask]
>Subject: Re: IMAGING OF CELLULOSE
>
>*****
>To join, leave or search the confocal microscopy listserv, go to:
>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>Post images on http://www.imgur.com and include the link in your posting.
>*****
>
>Scanning electron microscopy works very well for imaging papers and other
>such cellulosic products and their coatings.
>Also, the paper (etc.) can be "fractured" in liquid nitrogen to produce
>high quality edges to image the inner layers of the paper (etc.), and the
>coating layers. Cutting tends to smear the surface over the cut edge
>- this is more of an issue with coated papers.
>
>Phil
>
>> Hello,
>> Does anybody know of microscopy techniques for 3D imaging of paper or
>> other cellulose based fabrics? We have been trying with confocal
>> microscopy and did imaging of the surface of the outer layer of
>> cellulose fibers, but were unable to go through the fibers and image
>>the underlying layers.
>> Best regards,
>> Dario
>
>--
>Philip Oshel
>Microscopy Facility Supervisor
>Biology Department
>024C Brooks Hall
>Central Michigan University
>Mt. Pleasant, MI 48859
>(989) 774-3576
>
>
>
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