colocalization question

[Jean-Pierre CLAMME-2]

Hi,

When you do colocalization measurement (in image pro for example) between let say the red and a green channel the co-localization coefficient tells you how much red and green are co-localized.  But as much as I understood, it doesn't give you information on the fact that for example red is always co-localized with green but green is not always colocalized with red ? What kind of analysis  would I need to compare this between samples ?

Thank you in advance,

JP


- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
Jean-Pierre CLAMME, PhD
Senior Scientist
Nitto Denko Technical
501 Via Del Monte
Oceanside, CA 92058
E-mail: [hidden email]
Phone: [phone]

[Julio Vazquez]

-

Actually, many colocalization programs, including the imageJ plugin "Colocalization finder"  will tell you the percent of each channel that is colocalized. see for instance:

http://rsb.info.nih.gov/ij/plugins/colocalization-finder.html

--

Julio Vazquez

Fred Hutchinson Cancer Research Center

Seattle, WA 98109-1024

http://www.fhcrc.org/

On Feb 4, 2009, at 5:05 PM, Jean-Pierre CLAMME wrote:

Hi,

When you do colocalization measurement (in image pro for example) between let say the red and a green channel the co-localization coefficient tells you how much red and green are co-localized.  But as much as I understood, it doesn't give you information on the fact that for example red is always co-localized with green but green is not always colocalized with red ? What kind of analysis  would I need to compare this between samples ?

Thank you in advance,

JP


- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
Jean-Pierre CLAMME, PhD
Senior Scientist
Nitto Denko Technical
501 Via Del Monte
Oceanside, CA 92058
E-mail: [hidden email]
Phone: [phone]

[Jose Viña]

> But as much as I understood,
> it doesn't give you information on the fact that for example red is always
> co-localized with green but green is not always colocalized with red ? What
> kind of analysis  would I need to compare this between samples ?

Mander's coefficients are intended to give this kind of information,
see e.g. http://support.svi.nl/wiki/ColocalizationCoefficients
(coefficients interpretation at the end of the article).

Cheers,

jose.

[Yuri Gaidoukevitch]

Jean-Pierre CLAMME-2 wrote

When you do colocalization measurement (in image pro for example) between
let say the red and a green channel the co-localization coefficient tells
you how much red and green are co-localized.  But as much as I understood,
it doesn't give you information on the fact that for example red is always
co-localized with green but green is not always colocalized with red ?
What kind of analysis  would I need to compare this between samples ?

Jean-Pierre,

Image-Pro provides this information in m1/m2 and M1/M2 coefficients. You can find detailed information in http://www.mediacy.com/pdfs/colocfluorprobes.pdf   

Regards,

 

Yuri Gaidoukevitch
Senior Software Engineer

Media Cybernetics
Frambozenweg 139
2321KA Leiden

The Netherlands
tel  [phone]
mobile  [phone]
fax [phone]

 

[Jeremy Adler-2]

As Jose and Julio point out the data is readily available in the form you are looking for.

 

A wider question is what you take measurements to mean.

 

These coefficients need to be qualified by the area each fluorophore occupies in the ROI: if each occurred in 50% of the pixels, then chance alone would produce an overlap of 25% (uninteresting), while overlaps of say 10% or 40% would be more interesting – a non random mechanism.

If say they are both water soluble they might well appear throughout the cytoplasm and show 100% colocalization (using some variant of overlap).

Also bear in mind that these coefficients are very sensitive to the threshold used to decide if a fluorophore is present/absent in individual pixels.

 

A more interesting measure is correlation – does the variation in the intensity of one fluorophore match that of the second. This shows whether there is some underlying linkage between the distribution of the fluorophores. The variation would be caused by inhomegeneity in the volume of distribution. A correlation measurement for our two fluorophores that simply appear in the cytoplasm will be very low.

 

In practice use both overlap and correlation when considering colocalization and, as with all measurements, always consider exactly what has been measured and how it might be misleading.

 

 

 

Jeremy

 

 

Dr Jeremy Adler

F451a

Cell Biologi

Wenner-Gren Inst.

The Arhenius Lab

Stockholm University

S-106 91 Stockholm

Sweden

 

---

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Julio Vazquez
Sent: den 5 februari 2009 02:13
To: [hidden email]
Subject: Re: colocalization question

 

-

Actually, many colocalization programs, including the imageJ plugin "Colocalization finder" will tell you the percent of each channel that is colocalized. see for instance:

 

http://rsb.info.nih.gov/ij/plugins/colocalization-finder.html

--

Julio Vazquez

Fred Hutchinson Cancer Research Center

Seattle, WA 98109-1024

 

 

http://www.fhcrc.org/

 

 

On Feb 4, 2009, at 5:05 PM, Jean-Pierre CLAMME wrote:

Hi,

When you do colocalization measurement (in image pro for example) between let say the red and a green channel the co-localization coefficient tells you how much red and green are co-localized. But as much as I understood, it doesn't give you information on the fact that for example red is always co-localized with green but green is not always colocalized with red ? What kind of analysis would I need to compare this between samples ?

Thank you in advance,

JP


- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
Jean-Pierre CLAMME, PhD
Senior Scientist
Nitto Denko Technical
501 Via Del Monte
Oceanside, CA 92058
E-mail: [hidden email]
Phone: [phone]

 

[Daniel James White]

Dear JP,

You need to measure thresholded Manders coefficients (1 for red , 1  
for green!)

search this list archives for my previous long posts on this topic.


in short, use ImageJ or easier BioImageXD (both free)
to calculate threshold using the methods of costes,
then calculate the Manders coefficents 1 and 2
then do the costes statistical significance test.

Publish:
thresholds, manders coefficents, statistical significance,
2D histogram . scatter plots with straight line fit and thresholds.

You need to read 2 papers at least
  Costes et al
and Manders et al.

I will send them to you in a moment

cheers

Dan



Begin forwarded message:

>
> Date:    Wed, 4 Feb 2009 17:05:30 -0800
> From:    Jean-Pierre CLAMME <[hidden email]>
> Subject: colocalization question
>
> This is a multipart message in MIME format.
> --=_alternative 0005FF9888257554_=
> Content-Type: text/plain; charset="US-ASCII"
>
> Hi,
>
> When you do colocalization measurement (in image pro for example)  
> between
> let say the red and a green channel the co-localization coefficient  
> tells
> you how much red and green are co-localized.  But as much as I  
> understood,
> it doesn't give you information on the fact that for example red is  
> always
> co-localized with green but green is not always colocalized with red ?
> What kind of analysis  would I need to compare this between samples ?
>
> Thank you in advance,
>
> JP
>
>
> - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
> Jean-Pierre CLAMME, PhD
> Senior Scientist
> Nitto Denko Technical
> 501 Via Del Monte
> Oceanside, CA 92058
> E-mail: [hidden email]
> Phone: [phone]

Dr. Daniel James White BSc. (Hons.) PhD
Senior Microscopist / Image Processing and Analysis
Light Microscopy Facility
Max Planck Institute of Molecular Cell Biology and Genetics
Pfotenhauerstrasse 108
01307 DRESDEN
Germany


New Mobile Number!!!

[phone] (German Mobile)
+49  (0)351 210 2627 (Work phone at MPI-CBG)
+49  (0)351 210 1078 (Fax MPI-CBG LMF)

http://www.bioimagexd.net
http://www.chalkie.org.uk
[hidden email]
( [hidden email] )

[B. Prabhakar Pandian]

Hello,
           Does anybody have advice on what substrates
(Gelatin/Lysine/Collagen/Etc.)  is best for growing adherent cells on
glass slides. Is there a study where cell differentiation was compared
on various substrates.

Any help would be great.

Thanks,

-Prabhakar

[Jean-Pierre CLAMME-2]

Thank you all very much. Your inputs  helped a lot !

JP

- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
Jean-Pierre CLAMME, PhD
Senior Scientist
Nitto Denko Technical
501 Via Del Monte
Oceanside, CA 92058
E-mail: [hidden email]
Phone: [phone]


Confocal Microscopy List <[hidden email]> wrote on 02/05/2009 03:00:41 AM:

> Dear JP,

> You need to measure thresholded Manders coefficients (1 for red , 1
> for green!)

> search this list archives for my previous long posts on this topic.

>
> in short, use ImageJ or easier BioImageXD (both free)
> to calculate threshold using the methods of costes,
> then calculate the Manders coefficents 1 and 2
> then do the costes statistical significance test.

> Publish:
> thresholds, manders coefficents, statistical significance,
> 2D histogram . scatter plots with straight line fit and thresholds.

> You need to read 2 papers at least
> Costes et al
> and Manders et al.

> I will send them to you in a moment

> cheers

> Dan

>
> Begin forwarded message:

> >
> > Date:    Wed, 4 Feb 2009 17:05:30 -0800
> > From:    Jean-Pierre CLAMME <[hidden email]>
> > Subject: colocalization question
> >
> > This is a multipart message in MIME format.
> > --=_alternative 0005FF9888257554_=
> > Content-Type: text/plain; charset="US-ASCII"
> >
> > Hi,
> >
> > When you do colocalization measurement (in image pro for example)
> > between
> > let say the red and a green channel the co-localization coefficient
> > tells
> > you how much red and green are co-localized.  But as much as I
> > understood,
> > it doesn't give you information on the fact that for example red is
> > always
> > co-localized with green but green is not always colocalized with red ?
> > What kind of analysis  would I need to compare this between samples ?
> >
> > Thank you in advance,
> >
> > JP
> >
> >
> > - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
> > Jean-Pierre CLAMME, PhD
> > Senior Scientist
> > Nitto Denko Technical
> > 501 Via Del Monte
> > Oceanside, CA 92058
> > E-mail: [hidden email]
> > Phone: [phone]

> Dr. Daniel James White BSc. (Hons.) PhD
> Senior Microscopist / Image Processing and Analysis
> Light Microscopy Facility
> Max Planck Institute of Molecular Cell Biology and Genetics
> Pfotenhauerstrasse 108
> 01307 DRESDEN
> Germany

>
> New Mobile Number!!!

> [phone] (German Mobile)
> +49  (0)351 210 2627 (Work phone at MPI-CBG)
> +49  (0)351 210 1078 (Fax MPI-CBG LMF)

> http://www.bioimagexd.net
> http://www.chalkie.org.uk
> [hidden email]
> ( [hidden email] )

[Anchall ............]

Transfection microarray of human mesenchymal stem cells and on-chip siRNA gene knockdown

On Thu, Feb 5, 2009 at 5:56 PM, Anchall ............ <[hidden email]> wrote:

This study they have tested PLL ( poly L-Lysine),fibronectin and gelatin..

On Thu, Feb 5, 2009 at 5:47 PM, B. Prabhakar Pandian <[hidden email]> wrote:

Hello,
         Does anybody have advice on what substrates (Gelatin/Lysine/Collagen/Etc.)  is best for growing adherent cells on glass slides. Is there a study where cell differentiation was compared
on various substrates.

Any help would be great.

Thanks,

-Prabhakar

[Bowman, Doug]

Not specifically a confocal question:

 

Any opinions on commercially available media/drug perfusion system for multi-well plate live-cell imaging?  

 

We are looking for a system that would allow for media exchange / compound addition during a timelapse experiment without having to remove the plate from the environmental enclosure.  I am familiar with the Wafergen system and Fluxion BioFlux 200 products – but only from a web search.

 

Any opinions on either system or other companies would be great.

Thanks,
Doug

This e-mail, including any attachments, is a confidential business communication, and may contain information that is confidential, proprietary and/or privileged.  This e-mail is intended only for the individual(s) to whom it is addressed, and may not be saved, copied, printed, disclosed or used by anyone else.  If you are not the(an) intended recipient, please immediately delete this e-mail from your computer system and notify the sender.  Thank you.

[Kurt Thorn]

There is a system similar to the BioFlux from CellASIC
(http://www.cellasic.com/).

I've worked with both the BioFlux and the CellASIC and both work well
and are easy to use.

Kurt

Bowman, Doug wrote:

>
> Not specifically a confocal question:
>
> Any opinions on commercially available media/drug perfusion system for
> multi-well plate live-cell imaging?
>
> We are looking for a system that would allow for media exchange /
> compound addition during a timelapse experiment without having to
> remove the plate from the environmental enclosure. I am familiar with
> the Wafergen system and Fluxion BioFlux 200 products – but only from a
> web search.
>
> Any opinions on either system or other companies would be great.
>
>
> Thanks,
> Doug
>
>
>
> This e-mail, including any attachments, is a confidential business communication, and may contain information that is confidential, proprietary and/or privileged.  This e-mail is intended only for the individual(s) to whom it is addressed, and may not be saved, copied, printed, disclosed or used by anyone else.  If you are not the(an) intended recipient, please immediately delete this e-mail from your computer system and notify the sender.  Thank you.
>  

--
Kurt Thorn, PhD
Director, Nikon Imaging Center
University of California San Francisco

UCSF MC 2140
Genentech Hall Room S252
600 16th St.
San Francisco, CA 94158-2517

http://nic.ucsf.edu
phone [phone]
fax   [phone]

[Cameron Nowell]

Howdy,

          Sorry to chip in a bit late to this discussion but one thing you need to always remember with colocalisation of fluorecent images, that is resolution. Sorry to those that already know all this but is is an important point to emphasise when talking about colocalisation.

 

Resolution is governed by NA and can be calculated using 0.61 x emission wavlength/NA (for widfield) and 0.4 x emision wavelength/NA (for confocal). There are a few variations of this formula, and while they give theoretical best resolutions, the resolution acheived in practice is usually always worse. Also this is only for lateral resolution, axial resolution is worse by a facotr of about 3.

 

So for a 60x 1.4NA objective on a confocal your best lateral resolution for say GFP is about 150nm (220nm for widefield). That means that your proteins could be 200nm apart and still "colocalise". 200nm (2000 angstroms) is a rather large distance when you are talking about protein interactions. For example a human herpes virus is aorund 200nm across, there are a lot of proteins in that small particle, but not all of them are directly interating with each other.

 

Normally when people are doing colocalisation they have immunoprecipitation or other such data to show the proteins they are interested in interact, couple this with the fluorecent colocalisation and it is much more convincing. The other choice is FRET imaging, as FRET requires proteins to be 1-10nm appart for it to occur. So even though you can't resolve the protein with a confocal you can indirectly measure its interaction.

 

If you want an accurate image to show proteins interacting you have to use immuno EM as EM can give you the resolution you need.

 

Sorry for the rant :)

 

 

Cam

 

 

Cameron J. Nowell
Microscopy Manager
Central Resource for Advanced Microscopy
Ludwig Institute for Cancer Research
PO Box 2008
Royal Melbourne Hospital
Victoria, 3050
AUSTRALIA
Office: [phone]
Mobile: [phone]
Fax: [phone]
Facility Website

---

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Jean-Pierre CLAMME
Sent: Friday, 6 February 2009 3:57 AM
To: [hidden email]
Subject: Re: colocalization question

Thank you all very much. Your inputs  helped a lot !

JP

- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
Jean-Pierre CLAMME, PhD
Senior Scientist
Nitto Denko Technical
501 Via Del Monte
Oceanside, CA 92058
E-mail: [hidden email]
Phone: [phone]


Confocal Microscopy List <[hidden email]> wrote on 02/05/2009 03:00:41 AM:

> Dear JP,

> You need to measure thresholded Manders coefficients (1 for red , 1
> for green!)

> search this list archives for my previous long posts on this topic.

>
> in short, use ImageJ or easier BioImageXD (both free)
> to calculate threshold using the methods of costes,
> then calculate the Manders coefficents 1 and 2
> then do the costes statistical significance test.

> Publish:
> thresholds, manders coefficents, statistical significance,
> 2D histogram . scatter plots with straight line fit and thresholds.

> You need to read 2 papers at least
> Costes et al
> and Manders et al.

> I will send them to you in a moment

> cheers

> Dan

>
> Begin forwarded message:

> >

> > Date:    Wed, 4 Feb 2009 17:05:30 -0800
> > From:    Jean-Pierre CLAMME <[hidden email]>
> > Subject: colocalization question
> >
> > This is a multipart message in MIME format.
> > --=_alternative 0005FF9888257554_=
> > Content-Type: text/plain; charset="US-ASCII"
> >
> > Hi,
> >
> > When you do colocalization measurement (in image pro for example)
> > between
> > let say the red and a green channel the co-localization coefficient
> > tells
> > you how much red and green are co-localized.  But as much as I
> > understood,
> > it doesn't give you information on the fact that for example red is
> > always
> > co-localized with green but green is not always colocalized with red ?
> > What kind of analysis  would I need to compare this between samples ?
> >
> > Thank you in advance,
> >
> > JP
> >
> >
> > - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
> > Jean-Pierre CLAMME, PhD
> > Senior Scientist
> > Nitto Denko Technical
> > 501 Via Del Monte
> > Oceanside, CA 92058
> > E-mail: [hidden email]
> > Phone: [phone]

> Dr. Daniel James White BSc. (Hons.) PhD
> Senior Microscopist / Image Processing and Analysis
> Light Microscopy Facility
> Max Planck Institute of Molecular Cell Biology and Genetics
> Pfotenhauerstrasse 108
> 01307 DRESDEN
> Germany

>
> New Mobile Number!!!

> [phone] (German Mobile)
> +49  (0)351 210 2627 (Work phone at MPI-CBG)
> +49  (0)351 210 1078 (Fax MPI-CBG LMF)

> http://www.bioimagexd.net
> http://www.chalkie.org.uk
> [hidden email]
> ( [hidden email] )

---

This communication is intended only for the named recipient and may contain information that is confidential, legally privileged or subject to copyright; the Ludwig Institute for Cancer Research Ltd does not waiver any rights if you have received this communication in error.
The views expressed in this communication are those of the sender and do not necessarily reflect the views of the Ludwig Institute for Cancer Research Ltd.

[Jose Viña]

Cameron Nowell wrote:

> That means that your proteins could be 200nm apart and still "colocalise".

Some images to illustrate that can be found at
http://support.svi.nl/wiki/BlurAndNoiseAffectColocalization

Maybe Dr. Nowell wants to quote that email in that wiki article!

[Erik Manders-2]

Indeed, this kind of information is expressed by M1 and M2 (what the
community named the "Manders' coefficients"). However, be aware that you
should be very careful with your conclusions from your measurements. Like all
co-localization coefficients these coefficients are very sensitive for off-set
and cross-talk. A high offset gives a high background and background
correlates and may co-localize with backgound and this will strongly bias your
coefficient (especially in 3D data-sets since there are lot of background pixels
in 3D-images). Cross-talk is undistinguishable from co-localisation and should
therefore be reduced during scanning (sequential scanning) of removed by
post-processing. So you should allways measure your cross-talk in control
experiments (single colour labelling) with the same microscope settings (laser
power, pinhole, PMT voltage, etc) as your real experiment. Another point of
attention is the threshold that has to be set in the Manders' coefficients. This
may be a very critical setting, so be careful and do your control experiments.
The numbers that come out should be compared with these control
experiments. And this counts for all colocalisation coefficients. You should
compare the numbers with the numbers of control experiments. There are two
kind of experiments that you may do. 1) Biological control experiments: e.g. Do
another pair of stainings of which you know that they should colocalize and do
a pair of stainings of which you know that they cannot colocalize at all. Then
measure your relative "zero" and "one". 2) Image processing experiment (only
possible to measure your "zero"). Take two dual-colour images of look-alike
cells. Take the red component of the first and measure the colocalisation with
the green component of the second image. Repeat this for some more
combinations of images and see what the colocalisation coefficents are of two
uncorrelated patterns. It is clear that you apply the same post-processing
(cross-talk reduction, noise reduction, off-set reduction, deconvolution, etc)
and colocalisation measurement (thresholds, etc).
So, my take home message is: It is easy to get numbers from an image. The
interpretation of these numbers needs a lot of attention.

Good luck and succes with your research,
Erik Manders

[Renato A. Mortara]

Hi,

I would appreciate any input on the Leica Tandem Scanner confocal microscope
(pros and cons).

Many thanks !

Renato

Renato A. Mortara
Parasitology Division
UNIFESP - Escola Paulista de Medicina
Rua Botucatu, 862, 6th floor
São Paulo, SP
04023-062
Brazil
Phone: [phone]
Fax:     [phone]
email: [hidden email]
home page: www.ecb.epm.br/~ramortara
 

[Guy Cox]

Hello, what are we talking about here?  I've used the Leica STED
(and I'm seriously impressed with it) but I didn't know Leica made
a tandem scanner.  Please clarify!  I'm presenting results from
the Leica STED at NZCM next week in Rotorua and in April at FOM in
Krakow.  I do realise both are a bit remote from Brazil.

                                                       
                                                    Guy


Optical Imaging Techniques in Cell Biology
by Guy Cox    CRC Press / Taylor & Francis
    http://www.guycox.com/optical.htm
______________________________________________
Associate Professor Guy Cox, MA, DPhil(Oxon)
Electron Microscope Unit, Madsen Building F09,
University of Sydney, NSW 2006
______________________________________________
Phone [phone]     Fax [phone]
Mobile [phone]
______________________________________________
     http://www.guycox.net
-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Renato Mortara
Sent: Friday, 6 February 2009 9:37 PM
To: [hidden email]
Subject: Input on the Leica STED

Hi,

I would appreciate any input on the Leica Tandem Scanner confocal microscope (pros and cons).

Many thanks !

Renato

Renato A. Mortara
Parasitology Division
UNIFESP - Escola Paulista de Medicina
Rua Botucatu, 862, 6th floor
São Paulo, SP
04023-062
Brazil
Phone: [phone]
Fax:     [phone]
email: [hidden email]
home page: www.ecb.epm.br/~ramortara
 

No virus found in this incoming message.
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[simon walker (BI)]

Presumably we're talking about the SP5 with conventional and resonant scanners and adapted for STED..?

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Guy Cox
Sent: 06 February 2009 11:41
To: [hidden email]
Subject: Re: Input on the Leica STED

Hello, what are we talking about here?  I've used the Leica STED
(and I'm seriously impressed with it) but I didn't know Leica made
a tandem scanner.  Please clarify!  I'm presenting results from
the Leica STED at NZCM next week in Rotorua and in April at FOM in
Krakow.  I do realise both are a bit remote from Brazil.

                                                       
                                                    Guy


Optical Imaging Techniques in Cell Biology
by Guy Cox    CRC Press / Taylor & Francis
    http://www.guycox.com/optical.htm
______________________________________________
Associate Professor Guy Cox, MA, DPhil(Oxon)
Electron Microscope Unit, Madsen Building F09,
University of Sydney, NSW 2006
______________________________________________
Phone [phone]     Fax [phone]
Mobile [phone]
______________________________________________
     http://www.guycox.net
-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Renato Mortara
Sent: Friday, 6 February 2009 9:37 PM
To: [hidden email]
Subject: Input on the Leica STED

Hi,

I would appreciate any input on the Leica Tandem Scanner confocal microscope (pros and cons).

Many thanks !

Renato

Renato A. Mortara
Parasitology Division
UNIFESP - Escola Paulista de Medicina
Rua Botucatu, 862, 6th floor
São Paulo, SP
04023-062
Brazil
Phone: [phone]
Fax:     [phone]
email: [hidden email]
home page: www.ecb.epm.br/~ramortara
 

No virus found in this incoming message.
Checked by AVG.
Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: 4/02/2009 4:35 PM
 

No virus found in this outgoing message.
Checked by AVG.
Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: 4/02/2009 4:35 PM
 

[Michael Weber-4]

Hi, tandem scanner means you have a slider with both the galvano scanner
and a resonant scanner. The position of the slider (which scanner will be
used) can be choosen during system initation. So kind of like the Nikon
A1R, just that you cannot use both in parallel with the Leica SP5.

But this whole thing is independet from STED. So I don't know what we are
talking about :).

Michael

> Hello, what are we talking about here?  I've used the Leica STED
> (and I'm seriously impressed with it) but I didn't know Leica made
> a tandem scanner.  Please clarify!  I'm presenting results from
> the Leica STED at NZCM next week in Rotorua and in April at FOM in
> Krakow.  I do realise both are a bit remote from Brazil.
>
>
>                                                     Guy
>
>
> Optical Imaging Techniques in Cell Biology
> by Guy Cox    CRC Press / Taylor & Francis
>     http://www.guycox.com/optical.htm
> ______________________________________________
> Associate Professor Guy Cox, MA, DPhil(Oxon)
> Electron Microscope Unit, Madsen Building F09,
> University of Sydney, NSW 2006
> ______________________________________________
> Phone [phone]     Fax [phone]
> Mobile [phone]
> ______________________________________________
>      http://www.guycox.net
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]]
> On Behalf Of Renato Mortara
> Sent: Friday, 6 February 2009 9:37 PM
> To: [hidden email]
> Subject: Input on the Leica STED
>
> Hi,
>
> I would appreciate any input on the Leica Tandem Scanner confocal
> microscope (pros and cons).
>
> Many thanks !
>
> Renato
>
> Renato A. Mortara
> Parasitology Division
> UNIFESP - Escola Paulista de Medicina
> Rua Botucatu, 862, 6th floor
> São Paulo, SP
> 04023-062
> Brazil
> Phone: [phone]
> Fax:     [phone]
> email: [hidden email]
> home page: www.ecb.epm.br/~ramortara
>
>
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> Checked by AVG.
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> 4/02/2009 4:35 PM
>
>
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> Checked by AVG.
> Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date:
> 4/02/2009 4:35 PM

[Guy Cox]

Well, tandem scanning was the term coined by Mojimir Petran for
his spinning-disk confocal system.  Ever since then there have
been companies offering similar systems.  Leica used to offer
such a system (based on the BD CARV) but have since discontinued
it.  But it is something very different in all respects (including
both price and performance) from the Leica STED which is a super-
resolution system offering 85nm resolution.

                                                           Guy



Optical Imaging Techniques in Cell Biology
by Guy Cox    CRC Press / Taylor & Francis
    http://www.guycox.com/optical.htm
______________________________________________
Associate Professor Guy Cox, MA, DPhil(Oxon)
Electron Microscope Unit, Madsen Building F09,
University of Sydney, NSW 2006
______________________________________________
Phone [phone]     Fax [phone]
Mobile [phone]
______________________________________________
     http://www.guycox.net
-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of simon walker (BI)
Sent: Friday, 6 February 2009 10:46 PM
To: [hidden email]
Subject: Re: Input on the Leica STED

Presumably we're talking about the SP5 with conventional and resonant scanners and adapted for STED..?

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Guy Cox
Sent: 06 February 2009 11:41
To: [hidden email]
Subject: Re: Input on the Leica STED

Hello, what are we talking about here?  I've used the Leica STED (and I'm seriously impressed with it) but I didn't know Leica made a tandem scanner.  Please clarify!  I'm presenting results from the Leica STED at NZCM next week in Rotorua and in April at FOM in Krakow.  I do realise both are a bit remote from Brazil.

                                                       
                                                    Guy


Optical Imaging Techniques in Cell Biology
by Guy Cox    CRC Press / Taylor & Francis
    http://www.guycox.com/optical.htm
______________________________________________
Associate Professor Guy Cox, MA, DPhil(Oxon) Electron Microscope Unit, Madsen Building F09, University of Sydney, NSW 2006 ______________________________________________
Phone [phone]     Fax [phone]
Mobile [phone]
______________________________________________
     http://www.guycox.net
-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Renato Mortara
Sent: Friday, 6 February 2009 9:37 PM
To: [hidden email]
Subject: Input on the Leica STED

Hi,

I would appreciate any input on the Leica Tandem Scanner confocal microscope (pros and cons).

Many thanks !

Renato

Renato A. Mortara
Parasitology Division
UNIFESP - Escola Paulista de Medicina
Rua Botucatu, 862, 6th floor
São Paulo, SP
04023-062
Brazil
Phone: [phone]
Fax:     [phone]
email: [hidden email]
home page: www.ecb.epm.br/~ramortara
 

No virus found in this incoming message.
Checked by AVG.
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Checked by AVG.
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[Renato A. Mortara]

Hope it is slightly clearer...

Leica is offering the STED system that that can be an option to their TCS
SP5 confocal.

Besides, they also offer a tandem scanner for the same instrument,
presumably to achieve higher acquisitions speed.

Thanks again !

Renato

 

-----Mensagem original-----
De: Confocal Microscopy List [mailto:[hidden email]] Em
nome de simon walker (BI)
Enviada em: sexta-feira, 6 de fevereiro de 2009 09:46
Para: [hidden email]
Assunto: Re: Input on the Leica STED

Presumably we're talking about the SP5 with conventional and resonant
scanners and adapted for STED..?

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On
Behalf Of Guy Cox
Sent: 06 February 2009 11:41
To: [hidden email]
Subject: Re: Input on the Leica STED

Hello, what are we talking about here?  I've used the Leica STED (and I'm
seriously impressed with it) but I didn't know Leica made a tandem scanner.
Please clarify!  I'm presenting results from the Leica STED at NZCM next
week in Rotorua and in April at FOM in Krakow.  I do realise both are a bit
remote from Brazil.

                                                       
                                                    Guy


Optical Imaging Techniques in Cell Biology
by Guy Cox    CRC Press / Taylor & Francis
    http://www.guycox.com/optical.htm
______________________________________________
Associate Professor Guy Cox, MA, DPhil(Oxon) Electron Microscope Unit,
Madsen Building F09, University of Sydney, NSW 2006
______________________________________________
Phone [phone]     Fax [phone]
Mobile [phone]
______________________________________________
     http://www.guycox.net
-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On
Behalf Of Renato Mortara
Sent: Friday, 6 February 2009 9:37 PM
To: [hidden email]
Subject: Input on the Leica STED

Hi,

I would appreciate any input on the Leica Tandem Scanner confocal microscope
(pros and cons).

Many thanks !

Renato

Renato A. Mortara
Parasitology Division
UNIFESP - Escola Paulista de Medicina
Rua Botucatu, 862, 6th floor
São Paulo, SP
04023-062
Brazil
Phone: [phone]
Fax:     [phone]
email: [hidden email]
home page: www.ecb.epm.br/~ramortara
 

No virus found in this incoming message.
Checked by AVG.
Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: 4/02/2009
4:35 PM
 

No virus found in this outgoing message.
Checked by AVG.
Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: 4/02/2009
4:35 PM