Re: PSF bead shot but X Y and Z dont match

Posted by Matiar Jafari on
URL: http://confocal-microscopy-list.275.s1.nabble.com/PSF-bead-shot-but-X-Y-and-Z-dont-match-tp4589883p4589939.html

how would i then go about correcting the Z blur for all my other
images. if the XY resolution is just distorted because of the
limitations of optics? sorry im really new to this kind of stuff.
Thank You
-------
Matiar Jafari



On Wed, Feb 17, 2010 at 7:29 PM, Fred Mast <[hidden email]> wrote:

> Is this the diametre of your deconvolved bead? If it is not than you have
> diffraction to thank for the increased width. The .17 micron beads should be
> sub-resolution on your microscope and are representative of a point source
> of light. The resulting "blurred" image that you have captured is the result
> of the physical properties of light and the optical constraints of your
> microscope. A good source for further information can be found at the
> following links:
> http://support.svi.nl/wiki/SubResolution
> http://en.wikipedia.org/wiki/Diffraction
> But also try the Handbook for Biological Confocal Microscopy, James Pawley
> is the editor, for further information.
> Best,
> Fred
> On 17-Feb-10, at 8:14 PM, Matiar Jafari wrote:
>
> Hey Fellow ListServers,
>
> I purchased a PSF kit from Invitrogen and imaged the beads and when i
> loaded the beads to correct the Z blur i measured the XY of the bead
> to make sure the diameter matched but it turns out it doesnt.  The
> beads are supposedly .17um with +-.005 but whatever angle i measure
> the bead at in xy i keep getting .4 um.  Any ideas as to why this is
> happening?
>
> Thank You
> -------
> Matiar Jafari
>
>
> Fred D. Mast
> Department of Cell Biology
> Medical Sciences Building Room 5-14
> University of Alberta
> Edmonton, Alberta, T6G 2H7
> Canada
> Tel: [phone]
> [hidden email]
>
>
>
>