Posted by
G. Esteban Fernandez on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Cosmic-ray-particles-centenary-The-Chase-2012-tp7578803p7578815.html
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What I do for picrosirius red is sandwich the slide between two cir.
pol. consumer camera filters (oriented correctly and rotated to
extinction), no other polarizers or filters in place. I put one
somewhere between the light source and the specimen (e.g. on top of
the condenser) and the other somewhere between the specimen and the
camera/eyepieces (e.g. take out the fluor. filters and put it in
there). That worked on different microscopes (Nikon, Olympus, Zeiss)
with the filters at different points in the light path. It worked
with 10x and below (that's what the user needed) and I remember it not
working with 20x/0.8 but did with 20x/0.4, so maybe this approach
doesn't work at high NA, I don't know enough to tell you. If you try
it, maybe stop down the condenser aperture with high NA lenses?
-Esteban
On Fri, Aug 10, 2012 at 8:32 AM, Guy Cox <
[hidden email]> wrote:
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> The signal will always be weaker with CP light. But it will no longer be orientation dependent. See Pu Xu, Eleanor Kable, Colin J. R. Sheppard, and Guy Cox, 2010 A quasi-crystal model of collagen microstructure based on SHG microscopy. Chinese Optics Letters 8, 213-216
>
> Do be aware that your quarter wave plate must be quarter wave for your excitation wavelength - a visible light one will not work if you are using SH / 2P excitation. (Lambda/4 at 900 nm = lambda/2 at 450nm). The analyser is less critical.
>
> Guy
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:
[hidden email]] On Behalf Of Alberto Diaspro
> Sent: Thursday, 9 August 2012 11:01 PM
> To:
[hidden email]
> Subject: Re: Imaging collagen with circularly polarized light
>
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> maybe this set-up could help
> Diaspro, A., Bertolotto, M., Vergani, L., & Nicolini, C. (1991). Polarized light scattering of nucleosomes and polynucleosomes-in situ and in vitro studies. IEEE TRANSACTIONS ON BIOMEDICAL ENGINEERING, 38(7), 670-678.
>
>
> Il giorno 09/ago/2012, alle ore 14:15, Michael Abanto <
[hidden email]> ha scritto:
>
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>> Hi,
>> Does anybody have a good protocol for imaging collagen in embedded tissue, stained with picrosirius red? I had no problem imaging collagen with linear polarization, but I wanted to try circular and found it difficult.
>> I have an Olympus BX61 and a Nikon A1, as well as demo lambda plates for both. Do I need an analyzer specific for circular polarization? And do I need a linear polarizer before the circular - and how would you set that up on the Olympus?
>> Any tips or help on the set-up would be appreciated,
>> Mike