Re: Poor man's confocal - HiLo method
Posted by
lechristophe on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Re-Poor-man-s-confocal-HiLo-method-tp5112169p5112414.html
On Fri, May 28, 2010 at 10:31, Daniel James White
<[hidden email]> wrote:
Hi Christophe,
Begin forwarded message:
>
> Date: Thu, 27 May 2010 11:19:45 +0200
> From: Christophe Leterrier <[hidden email]>
> Subject: Re: Poor man's confocal
>
> There is at least another cheap and smart method to get optical
> sectioning from a widefield microscope, called "HiLo" microscopy:
>
> Wide-field fluorescence sectioning with hybrid speckle and
> uniform-illumination microscopy.
> Lim D, Chu KK, Mertz J.
> Opt Lett. 2008 Aug 15;33(16):1819-21.
>
> The idea is to take a speckled image and a non speckled one, from
> which high and low frequencies of the images are computed and merged
> to from the final image.
looks very simple.... i wonder if you really need a laser illumination ? Would a metal halide lamp also work?
I dont see why not? Ot does it need to be a coherent beam?
I'm not sure but I think you need to form a speckled illumination pattern. Does this requires a coherent beam ?
> I don't think there is a commercial version
> of it, but I've seen two other labs using it and it seems quite simple
> to implement.
do you know how what kind of diffuser they use and where to get one?
Don't laugh, but the setup I've seen working used a bit of transparent
(invisible "crystal") adhesive tape as the diffuser...
how did they move the diffuser fast without vibrations?
I think the adhesive tape bit was
just put in a motorized filter wheel or something like that.
cheers
Dan
Dr. Daniel James White BSc. (Hons.) PhD
Senior Microscopist / Image Visualisation, Processing and Analysis
Light Microscopy and Image Processing Facilities
Max Planck Institute of Molecular Cell Biology and Genetics
Pfotenhauerstrasse 108
01307 DRESDEN
Germany
+49 (0)15114966933 (German Mobile)
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( white (at) mpi-cbg.de )
Cheers,
Christophe