Posted by
Mark Cannell on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Deep-tissue-imaging-with-spinning-disc-comparison-with-other-confocal-techniques-tp7561610p7565413.html
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The improved penetration of SDC compared to confocal is almost certainly the wider pinhole of the SDC compared to confocal IMHO. As a shameless self citation, I think that the penetration depth issue and effect of pinhoie etc. are quite well illustrated in our paper:
Soeller C, Cannell MB. Two-photon microscopy: Imaging in scattering samples and three-dimensionally resolved flash photolysis. Microsc. Res. Tech. 1999;47(3):182–95.
Note the effect of opening the pinhole fully in our Fig. 4 and the relative performance loss due to closing the pinhole with depth (highly non-linear with essentially no signal beyond 50 um in this sample). In this strongly scattering sample (cheese), 1% signal was obtained at 45 um with a pinhole, opening the pinhole increased the signal ~10x and using non-descanned detection increased the signal ~3x more. The method used 2P excitation in all cases so that we could examine the detector side performance -so the differences would be even greater for 1P where scattering also decreases excitation intensity at the focal spot ...
Cheers Mark
On 18/05/2012, at 8:10 AM, Sebastian Rhode wrote:
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>
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>
> Hi Jan,
>
> since our company manufactures both kind of systems, spinning disc confocal
> and 2-Photon systems, just my opinion on this paper.
>
> What I have seen so far during our tests, shows a clear advantage of 2PM if
> depth penetration is your primary(!) goal. If it comes to speed, spinning
> disc confocal (SDC) systems have a clear advantage. And in terms of cell
> viability, 2PM and SDC are better than a conventional LSM microscope.
>
> What puzzles me a bit, is the comparison between the penetration depth of
> the SDC and LSM. There are several points (some of them already mentioned by
> George here), which makes it difficult for me to really compare the results.
>
> But nevertheless the findings are quite interesting, even if there are open
> questions to answer. But I would be really interested in the explanation
> about the observed results.
>
> Dr. Sebastian Rhode
> Project Manager Research & Development
>
> TILL Photonics GmbH
> an FEI Company