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The equations I was thinking about in this book chapter are the ones in
paragraphs 2.3 and 3.3. No a priori assumption is made there about pinhole
sizes. They rather give the general formalism how to calculate effective
PSFs and the optical sectioning capability. I found this to be very
instructive. As I said in my previous message, the finite pinhole size can
be taken into account by calculating the convolution product of the pinhole
with the objective's PSF. This would be the effective PSF at the detection
side (h_det in the book chapter).

Sorry, don't know about the calculations for a spinning disk system.

Best regards,

Kevin



Kevin Braeckmans, Ph.D.
Lab. General Biochemistry and Physical Pharmacy
Ghent University
Harelbekestraat 72
9000 Ghent
Belgium
Tel: +32 (0)9 264.80.78
Fax: +32 (0)9 264.81.89
> -----Oorspronkelijk bericht-----
> Van: Confocal Microscopy List [mailto:[log in to unmask]]
> Namens David Knecht-charter
> Verzonden: donderdag 11 oktober 2007 3:27
> Aan: [log in to unmask]
> Onderwerp: Re: pinhole and voxel
> 
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
> 
> The equations being used here don't take into account the size of the
> excitation and emission pinholes.  This can vary at the emission end
> for an LSCM.  At the excitation end I don't know if all the
> instruments are the same.  For the spinning disks, how do you compare
> the resolution/depth of field of a nipkow spinning disk, with a CARV/
> Leica type device which have different pinhole sizes for excitation
> and emission? Am I correct that the laser vs. non-laser of the two
> spinning disks makes no difference?  Dave
> 
> Dr. David Knecht
> Department of Molecular and Cell Biology
> Co-head Flow Cytometry and Confocal Microscopy Facility
> U-3125
> 91 N. Eagleville Rd.
> University of Connecticut
> Storrs, CT 06269
> 860-486-2200
> 860-486-4331 (fax)
> 
> 
> On Oct 10, 2007, at 3:47 AM, Kevin Braeckmans wrote:
> 
> > Search the CONFOCAL archive at
> > http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
> >
> > Dear Max,
> >
> > Maybe some of the more knowledgeable scientists on this list may
> > give you a
> > more detailed answer, but as far as I understand it is the same back
> > projection of the pinhole that you need to calculate the axial
> > resolution.
> >
> > If you know the 3D PSF of the objective lens, you can calculate the
> > backprojection of the pinhole in 3D, which is the convolution
> > product of the
> > pinhole with the PSF (which is the PSF itself for an infinitely small
> > pinhole). The total confocal resolution can then be calculated as the
> > product of this backprojected pinhole distribution and the
> > objective's PSF.
> >
> > There is a very comprehensive chapter on the fundamentals of
> > resolution of
> > different types of microscopes (confocal, widefield and Multi-
> > photon) in
> > Alberto Diaspro's excellent handbook:
> >
> > J. E. N. Jonkman and E. H. K. Stelzer. Resolution and contrast in
> > confocal
> > and two-photon microscopy.
> > In A. Diaspro, editor, Confocal and Two-Photon Microscopy:
> > Foundations,
> > Applications and Advances, book chapter 5, pages 101–125. Wiley-
> > Liss, Inc.,
> > New York, USA, 2002.
> >
> >
> > I really recommend reading this chapter. It is very clearly written
> > and also
> > explains the difference between optical sectioning and axial
> > resolution.
> >
> > Best regards,
> >
> > Kevin
> >
> >
> > Kevin Braeckmans, Ph.D.
> > Lab. General Biochemistry & Physical Pharmacy
> > Ghent University
> > Harelbekestraat 72
> > 9000 Ghent
> > Belgium
> > Tel: +32 (0)9 264.80.78
> > Fax: +32 (0)9 264.81.89
> > E-mail: [log in to unmask]
> >
> >
> >> -----Oorspronkelijk bericht-----
> >> Van: Confocal Microscopy List [mailto:[log in to unmask]]
> >> Namens [log in to unmask]
> >> Verzonden: dinsdag 9 oktober 2007 23:57
> >> Aan: [log in to unmask]
> >> Onderwerp: pinhole and voxel
> >>
> >> Search the CONFOCAL archive at
> >> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
> >>
> >> Dear All,
> >>
> >> I'm quite new in the confocal world, I'll start a PhD and I will
> have
> >> to use confocal.
> >> I well understand the principle of pinhole and I can figure out its
> >> effect on XY resolution but I can't figure out precisely what
> >> effect it
> >> have about Z resolution.
> >> If the pinhole leads to have 1 AU, what will be the z resolution ?
> >> How
> >> to calculate it ?
> >> Knowing the pinhole size we can calculte XY resolution by calculated
> >> the backprojected pinhole radius but how this value is related to z
> >> resolution ?
> >>
> >> Thanks for help
> >>
> >> Max
> >>
> >> ------------------------ ALICE C'EST ENCORE MIEUX AVEC CANAL+ LE
> >> BOUQUET ! ---------------
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> >> Soumis à conditions.