*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

/Commercial response/

Dear Brian,

The Huygens software team has recently developed a STED deconvolution 
option that works extremely well.

Have a look at http://www.svi.nl/STED

I'll be happy to give you off-line more background information why 
deconvolution of STED data is so successful.


With kind wishes,

Gitta Hamel

/--
Managing Director Huygens SVI tel: +31 (0)35 642 16 26 fax: +31 (0)35 
683 79 71 skype: gittahamel cell: +31(0)618 021272 Visiting address 
Laapersveld 63, 1213 VB Hilversum, The Netherlands /


On 08/31/2012 05:22 PM, Brian Northan wrote:
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> On a related note can anybody recommend a paper that explains how to
> calculate the STED PSF?
>
> Is it worth it to deconvolve this type of data? I've heard conflicting reports.
>
> On Fri, Aug 31, 2012 at 9:56 AM, Martin Wessendorf <[log in to unmask]> wrote:
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>>
>> Dear Alan--
>>
>>
>> On 8/31/2012 7:22 AM, Alan Smith wrote:
>>
>>> I understand how the illumination psf is formed in STED using a depletion
>>> beam. However, if the emission is detected is collected in epi-detection,
>>> why is the resolution not determined by the diffraction limit for the
>>> objective.
>>>
>>> As an example, if a single molecule was producing fluorescence, the image
>>> will be an airy disk determined by the NA of the detecting objective and
>>> the
>>> wavelength of light. Despite the fluorescence solely coming from a single
>>> molecule.
>>
>> You're right: with both a single molecule and with STED, you see a
>> diffraction-limited Airy disk on the emission side.  However, with STED, you
>> know to a high degree of precision the position of the excitation beam
>> eliciting that Airy disk.  Thus, as you scan that small excitation beam over
>> a small structure, you can obtain resolution that's many times better than
>> confocal.  In that way, it's similar to near-field super-resolution methods.
>>
>> Hope that helps!
>>
>> Martin Wessendorf
>> --
>> Martin Wessendorf, Ph.D.                   office: (612) 626-0145
>> Assoc Prof, Dept Neuroscience                 lab: (612) 624-2991
>> University of Minnesota             Preferred FAX: (612) 624-8118
>> 6-145 Jackson Hall, 321 Church St. SE    Dept Fax: (612) 626-5009
>> Minneapolis, MN  55455                    e-mail: [log in to unmask]

-- 
Managing Director

Huygens SVI

tel: +31 (0)35 642 16 26

fax:  +31 (0)35 683 79 71

skype: gittahamel

cell: +31(0)618 021272

Visiting address

Laapersveld 63,
1213 VB Hilversum,
The Netherlands