7th Course on Optical Microscopy Imaging for Biosciences | 20-24 April 2015 | IBMC, Porto

[Paula Sampaio]

*****
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http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

*****
Dear all,

The deadline for the registration in the course was extended until *28th
July*.

Looking forward to seeing you in Porto
<http://www.visitporto.travel/Visitar/Paginas/default.aspx>.


Best regards,

Paula Sampaio


**I Course Leica on Advance Confocal Microscopy and STED super resolution**

7th-9th July 2015 | IBMC | Porto | Portugal


Leica will organize in collaboration with IBMC a course Leica on Advanced
Confocal Microscopy and STED Super Resolution. This course will be from 7th
to 9th July of 2015 at IBMC and aims to provide extend technical skills and
know-how in advanced confocal imaging methods and in the state of art Leica
equipment including the STED super resolution, Light Sheet microscopy
(DLS), high content analysis and widefield FRAP. It combines introductory
lectures and practical hands-on sessions. During the practical sessions,
students will be able to observe their own samples.

LECTURERS
Nathalie Garin, Leica Microsystems (Schweiz) AG
Clément Laigle, Leica Microsistemas S.L.
Javier Pazos, Leica Microsistemas S.L
Paco Porto, Leica Microsistemas S.L.
Paula Sampaio, Advanced Light Microscopy, IBMC

INVITED SPEAKERS
José Rino Henriques, Bioimaging Unit, IMM
Renato Soccodato, Glial Cell Group, IBMC
Fábio Fernandes, Instituto Superior Técnico


More information at
https://www.ibmc.up.pt/training/workshops/i-course-leica-advance-confocal-microscopy-and-sted-super-resolution


Registration fee: 350 Euros (includes documentation, course material,
lunches, coffee breaks and Course Dinner)

Those interested should apply by filling the pre-registration form online
at: https://www.ibmc.up.pt/eventos/registrationnew.php?eid=76


--
***********************************************************

Paula Sampaio, PhD.

IBMC - Instituto de Biologia Molecular e Celular
Universidade do Porto
Rua Campo Alegre 823
P4150-180 PORTO
PORTUGAL

Tel: [phone] Ext: 1606
Fax: [phone]

e-mail: [hidden email]

Web:
http://www.ibmc.up.pt
https://www.ibmc.up.pt/research/research-facilities/advanced-light-microscopy/home

PPBI - Portuguese Platform of BioImaging
http://www.ppbi.pt

***********************************************************


--
***********************************************************

Paula Sampaio, PhD.

IBMC - Instituto de Biologia Molecular e Celular
Universidade do Porto
Rua Campo Alegre 823
P4150-180 PORTO
PORTUGAL

Tel: [phone] Ext: 1606
Fax: [phone]

e-mail: [hidden email]

Web:
http://www.ibmc.up.pt
https://www.ibmc.up.pt/research/research-facilities/advanced-light-microscopy/home

PPBI - Portuguese Platform of BioImaging
http://www.ppbi.pt

***********************************************************

[Paula Sampaio]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Dear all,

Please note that the deadline for the registration in the course was
extended until *28th JUNE* and not July as was indicated in previous
announcement.

I apologize for the mistake and duplication of e-mail.

Looking forward to seeing you in Porto
<http://www.visitporto.travel/Visitar/Paginas/default.aspx>.


Best regards,

Paula Sampaio



**I Course Leica on Advance Confocal Microscopy and STED super resolution**

7th-9th July 2015 | IBMC | Porto | Portugal


Leica will organize in collaboration with IBMC a course Leica on Advanced
Confocal Microscopy and STED Super Resolution. This course will be from 7th
to 9th July of 2015 at IBMC and aims to provide extend technical skills and
know-how in advanced confocal imaging methods and in the state of art Leica
equipment including the STED super resolution, Light Sheet microscopy
(DLS), high content analysis and widefield FRAP. It combines introductory
lectures and practical hands-on sessions. During the practical sessions,
students will be able to observe their own samples.

LECTURERS
Nathalie Garin, Leica Microsystems (Schweiz) AG
Clément Laigle, Leica Microsistemas S.L.
Javier Pazos, Leica Microsistemas S.L
Paco Porto, Leica Microsistemas S.L.
Paula Sampaio, Advanced Light Microscopy, IBMC

INVITED SPEAKERS
José Rino Henriques, Bioimaging Unit, IMM
Renato Soccodato, Glial Cell Group, IBMC
Fábio Fernandes, Instituto Superior Técnico


More information at
https://www.ibmc.up.pt/training/workshops/i-course-leica-advance-confocal-microscopy-and-sted-super-resolution 



Registration fee: 350 Euros (includes documentation, course material,
lunches, coffee breaks and Course Dinner)

Those interested should apply by filling the pre-registration form online
at: https://www.ibmc.up.pt/eventos/registrationnew.php?eid=76


--
***********************************************************

Paula Sampaio, PhD.

IBMC - Instituto de Biologia Molecular e Celular
Universidade do Porto
Rua Campo Alegre 823
P4150-180 PORTO
PORTUGAL

Tel: [phone] Ext: 1606
Fax: [phone]

e-mail: [hidden email]

Web:
http://www.ibmc.up.pt
https://www.ibmc.up.pt/research/research-facilities/advanced-light-microscopy/home

PPBI - Portuguese Platform of BioImaging
http://www.ppbi.pt

***********************************************************

[Vitaly Boyko]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Dear List,
I would greatly appreciate if someone could shed light on the extent of lateral and axial chromatic aberrations correction, and its limitations in the VIS portion of spectrum when image has been projected into the non-descanned detectors (one and two photon imaging)?
Many thanks in advance.
Vitaly
 


   

[Craig Brideau]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Your question is fairly vague, but here are some general answers: With 2
photon the important thing is how well the laser is focused. This means
your objective has to be well corrected for the NIR beam from the laser.
The resulting visible fluorescence is directed towards wide-aperture PMTs,
which don't care about aberration very much.
For single-photon confocal, you have to get your fluorescence through the
confocal pinhole, which will be *very* picky about aberration. Since you
are trying to get the light through a very small aperture any aberration
will cause signal loss.

Craig

On Mon, Jun 29, 2015 at 4:04 PM, Vitaly Boyko <[hidden email]>
wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear List,
> I would greatly appreciate if someone could shed light on the extent of
> lateral and axial chromatic aberrations correction, and its limitations in
> the VIS portion of spectrum when image has been projected into the
> non-descanned detectors (one and two photon imaging)?
> Many thanks in advance.
> Vitaly
>
>
>
>
>

[Vitaly Boyko]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Hi Craig,
thank you for your reply. How would one image Alexa 790/Cy7 together with let say CeFP/dapi, GFP through the one photon confocal system (with Perkin Elmer APDs as detectors). Are there many IR APOCHROMAT lens that are corrected between 400 nm (tough, I guess, at least 500 nm) and 900nm at 40x NA 1.0-1.1 or 60x NA 1.2?Multi-photon collect scattered (non-linear, non-chroma corrected) light via NDDs...
Vitaly 


     On Monday, June 29, 2015 6:16 PM, Craig Brideau <[hidden email]> wrote:
   

 Your question is fairly vague, but here are some general answers: With 2 photon the important thing is how well the laser is focused. This means your objective has to be well corrected for the NIR beam from the laser. The resulting visible fluorescence is directed towards wide-aperture PMTs, which don't care about aberration very much.For single-photon confocal, you have to get your fluorescence through the confocal pinhole, which will be *very* picky about aberration. Since you are trying to get the light through a very small aperture any aberration will cause signal loss.
Craig
On Mon, Jun 29, 2015 at 4:04 PM, Vitaly Boyko <[hidden email]> wrote:

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Dear List,
I would greatly appreciate if someone could shed light on the extent of lateral and axial chromatic aberrations correction, and its limitations in the VIS portion of spectrum when image has been projected into the non-descanned detectors (one and two photon imaging)?
Many thanks in advance.
Vitaly

[Craig Brideau]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Ah, you are using far red dyes with a confocal. That's a relatively new
thing. In this case the NIR corrected lenses normally used for 2-photon are
actually useful in a confocal situation since you are working with very red
light both for excitation and emission. If you also want to image GFP then
your system has to be very broadly corrected.
Also, hopefully someone can chime in here, how sensitive are the Perkin
Elmer APDs to 800 to 900nm light? Many detectors are very insensitive this
far down. You typically want to use a multi-alkali PMT for those sorts of
ranges. I find the Hamamatsu 10721-20 works well out to about 850-900nm yet
is still sensitive 'enough' in the blue and green as well (I use it for GFP
or DAPI and very red things). Here's the product sheet:
http://www.hamamatsu.com/eu/en/product/alpha/P/3003/3044/H10721-20/index.html
Getting back to your objective lens, in this case you need something that
is very broadly chromatically corrected. You will want to consult your
vendor as to which lenses they offer that meet this need. You will also
want to avoid working on the edge of your field of view, and might have to
crop down a bit as chromatic aberration gets worse near the edge of your
full field.

Best of luck!

Craig Brideau


On Mon, Jun 29, 2015 at 4:27 PM, Vitaly Boyko <[hidden email]>
wrote:

> Hi Craig,
>
> thank you for your reply. How would one image Alexa 790/Cy7 together with
> let say CeFP/dapi, GFP through the one photon confocal system (with Perkin
> Elmer APDs as detectors). Are there many IR APOCHROMAT lens that are
> corrected between 400 nm (tough, I guess, at least 500 nm) and 900nm at 40x
> NA 1.0-1.1 or 60x NA 1.2?Multi-photon collect scattered (non-linear,
> non-chroma corrected) light via NDDs...
>
> Vitaly
>
>
>
>
>   On Monday, June 29, 2015 6:16 PM, Craig Brideau <[hidden email]>
> wrote:
>
>
> Your question is fairly vague, but here are some general answers: With 2
> photon the important thing is how well the laser is focused. This means
> your objective has to be well corrected for the NIR beam from the laser.
> The resulting visible fluorescence is directed towards wide-aperture PMTs,
> which don't care about aberration very much.
> For single-photon confocal, you have to get your fluorescence through the
> confocal pinhole, which will be *very* picky about aberration. Since you
> are trying to get the light through a very small aperture any aberration
> will cause signal loss.
>
> Craig
>
> On Mon, Jun 29, 2015 at 4:04 PM, Vitaly Boyko <[hidden email]>
> wrote:
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear List,
> I would greatly appreciate if someone could shed light on the extent of
> lateral and axial chromatic aberrations correction, and its limitations in
> the VIS portion of spectrum when image has been projected into the
> non-descanned detectors (one and two photon imaging)?
> Many thanks in advance.
> Vitaly
>
>
>
>
>
>
>
>

[Michael Giacomelli]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

As Craig says, for two photon imaging with non-descanned detectors, only
the chromatic aberration of the excitation (in the NIR) matters.  This can
be quite severe as most objectives (even NIR enhanced ones) are
not achromatized in the NIR, and so can have very large chromatic
aberration for multiphoton applications.  Zeiss, for instance, recommended
us an NIR enhanced 0.45 NA objective that measured a gigantic 100 nm of
chromatic focal shift per 1 nm shift of center wavelength from 700-900 nm.
Such an objective will work fine with typical 150 fs excitation, but is
almost useless for shorter pulses or confocal imaging in the NIR.

If you think you need good broadband NIR-vis achromatization (and most
people don't really) then I would strongly recommend you request an
evaluation objective from the vendor to test before you buy it.  Many
vendors will happily recommend NIR objectives that are actually quite poor
in the NIR.

Mike

On Mon, Jun 29, 2015 at 6:44 PM, Craig Brideau <[hidden email]>
wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Ah, you are using far red dyes with a confocal. That's a relatively new
> thing. In this case the NIR corrected lenses normally used for 2-photon are
> actually useful in a confocal situation since you are working with very red
> light both for excitation and emission. If you also want to image GFP then
> your system has to be very broadly corrected.
> Also, hopefully someone can chime in here, how sensitive are the Perkin
> Elmer APDs to 800 to 900nm light? Many detectors are very insensitive this
> far down. You typically want to use a multi-alkali PMT for those sorts of
> ranges. I find the Hamamatsu 10721-20 works well out to about 850-900nm yet
> is still sensitive 'enough' in the blue and green as well (I use it for GFP
> or DAPI and very red things). Here's the product sheet:
>
> http://www.hamamatsu.com/eu/en/product/alpha/P/3003/3044/H10721-20/index.html
> Getting back to your objective lens, in this case you need something that
> is very broadly chromatically corrected. You will want to consult your
> vendor as to which lenses they offer that meet this need. You will also
> want to avoid working on the edge of your field of view, and might have to
> crop down a bit as chromatic aberration gets worse near the edge of your
> full field.
>
> Best of luck!
>
> Craig Brideau
>
>
> On Mon, Jun 29, 2015 at 4:27 PM, Vitaly Boyko <[hidden email]>
> wrote:
>
> > Hi Craig,
> >
> > thank you for your reply. How would one image Alexa 790/Cy7 together with
> > let say CeFP/dapi, GFP through the one photon confocal system (with
> Perkin
> > Elmer APDs as detectors). Are there many IR APOCHROMAT lens that are
> > corrected between 400 nm (tough, I guess, at least 500 nm) and 900nm at
> 40x
> > NA 1.0-1.1 or 60x NA 1.2?Multi-photon collect scattered (non-linear,
> > non-chroma corrected) light via NDDs...
> >
> > Vitaly
> >
> >
> >
> >
> >   On Monday, June 29, 2015 6:16 PM, Craig Brideau <
> [hidden email]>
> > wrote:
> >
> >
> > Your question is fairly vague, but here are some general answers: With 2
> > photon the important thing is how well the laser is focused. This means
> > your objective has to be well corrected for the NIR beam from the laser.
> > The resulting visible fluorescence is directed towards wide-aperture
> PMTs,
> > which don't care about aberration very much.
> > For single-photon confocal, you have to get your fluorescence through the
> > confocal pinhole, which will be *very* picky about aberration. Since you
> > are trying to get the light through a very small aperture any aberration
> > will cause signal loss.
> >
> > Craig
> >
> > On Mon, Jun 29, 2015 at 4:04 PM, Vitaly Boyko <[hidden email]>
> > wrote:
> >
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > Post images on http://www.imgur.com and include the link in your
> posting.
> > *****
> >
> > Dear List,
> > I would greatly appreciate if someone could shed light on the extent of
> > lateral and axial chromatic aberrations correction, and its limitations
> in
> > the VIS portion of spectrum when image has been projected into the
> > non-descanned detectors (one and two photon imaging)?
> > Many thanks in advance.
> > Vitaly
> >
> >
> >
> >
> >
> >
> >
> >
>