Producing Large volume photodamage

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Jeremy Adler Jeremy Adler
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Producing Large volume photodamage

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We are interested in photodamage and want to reproduce the power levels, achieved in the tiny volume under a microscope objective, over a whole petri dish.
 
We hope to obtain sufficient material to undertake chemical analysis.
 
We are considering a lecture theatre projector and using uniform coloured images to produce some wavelength specificity.
 
Any experiences, suggestions or advice about would be appreciated, either on or off line.
 
 
Jeremy Adler
Cell Biology
The Wenner-Gren Inst.
Arrhenius Laboratories E5
Stockholm University
Stockholm 106 91
Sweden
 
lechristophe lechristophe
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Re: Producing Large volume photodamage

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
I recently read an article in Nature Methods (Shaner et al. from R. Tsien lab) dealing with photostability of orange and red fluorescent proteins :
http://www.ncbi.nlm.nih.gov/pubmed/18454154

They used a macrosopic illuminator to bleach whole Petri dishes of bacteria expressing the fluorescent proteins, I think you can get more details in the Material & Methods.

Christophe



On Thu, Aug 21, 2008 at 10:34, Jeremy Adler <[hidden email]> wrote:
Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
We are interested in photodamage and want to reproduce the power levels, achieved in the tiny volume under a microscope objective, over a whole petri dish.
 
We hope to obtain sufficient material to undertake chemical analysis.
 
We are considering a lecture theatre projector and using uniform coloured images to produce some wavelength specificity.
 
Any experiences, suggestions or advice about would be appreciated, either on or off line.
 
 
Jeremy Adler
Cell Biology
The Wenner-Gren Inst.
Arrhenius Laboratories E5
Stockholm University
Stockholm 106 91
Sweden
 

Beat Ludin Beat Ludin
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Re: Producing Large volume photodamage

In reply to this post by Jeremy Adler
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

If you need a lot of power, you might want to consider OBB's KiloArc
lamp (http://www.obb1.com/LightSources/KiloArc.html).
Just something I have come across recently, no personal experience
and no commercial interest.

Beat


At 10:34 21-08-2008, you wrote:

>Search the CONFOCAL archive at
>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>We are interested in photodamage and want to reproduce the power
>levels, achieved in the tiny volume under a microscope objective,
>over a whole petri dish.
>
>We hope to obtain sufficient material to undertake chemical analysis.
>
>We are considering a lecture theatre projector and using uniform
>coloured images to produce some wavelength specificity.
>
>Any experiences, suggestions or advice about would be appreciated,
>either on or off line.
>
>
>Jeremy Adler
>Cell Biology
>The Wenner-Gren Inst.
>Arrhenius Laboratories E5
>Stockholm University
>Stockholm 106 91
>Sweden
>
><mailto:[hidden email]>[hidden email]
Koo, Lily (NIH/NIAID) [E] Koo, Lily (NIH/NIAID) [E]
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Intensity Gaussian

In reply to this post by lechristophe
Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Hello all,

 

My colleague would like to construct Gaussian curves for pixel intensities for individual cells in his acquired images.  Do you know if any software (e.g. Image J plug-ins) has such a feature?

 

Thanks,

 

Lily

Peter Zoon Peter Zoon
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Re: Intensity Gaussian

Search the CONFOCAL archive at
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In ImageJ you can draw a line on your image and then plot the intensity profile along that line.
The profile can then either be saved as image or exported as x,y pair into other software like, Igor Pro, Origin or even Excel.

-----Oorspronkelijk bericht-----
Van: Confocal Microscopy List namens Koo, Lily (NIH/NIAID) [F]
Verzonden: vr 8/22/2008 9:06
Aan: [hidden email]
Onderwerp: Intensity Gaussian
 
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Hello all,

 

My colleague would like to construct Gaussian curves for pixel
intensities for individual cells in his acquired images.  Do you know if
any software (e.g. Image J plug-ins) has such a feature?

 

Thanks,

 

Lily
G. Esteban Fernandez G. Esteban Fernandez
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Re: Intensity Gaussian

In reply to this post by Koo, Lily (NIH/NIAID) [E]
Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
Lily,
 
ImageJ can fit a Gaussian curve with the Analyze > Tools > Curve fitting... command.  Make sure you have the latest version (1.41j).  When you run that function you can paste (Ctrl-v) the numbers you wish to fit into the space provided.
 
For example, to fit the frequency of intensities within a cell:
1. Outline the cell with some drawing tool.
2. Press Ctrl-h to display the histogram.
3. Click the "Copy" button in the histogram window to copy the values to the clipboard.
4. Run Analyze > Tools > Curve fitting...
5. Select "Gaussian"
6. Erase the default numbers and press Ctrl-v to paste the histogram numbers you previously copied to the clipboard.
7. Click the "Fit" button.
8. A window will appear with the curve fitting.  You can save the curve coordinates as a text file with the "Save" button.  You can also List the numbers to see them or you can Copy them to the clipboard.
 
I don't know if this is exactly what you need to do, but I hope at least the Curve Fitting function helps you in some way.  Contact me off list if you have questions.
 
-Esteban
 
 

 
On Fri, Aug 22, 2008 at 2:06 PM, Koo, Lily (NIH/NIAID) [F] <[hidden email]> wrote:
Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Hello all,

 

My colleague would like to construct Gaussian curves for pixel intensities for individual cells in his acquired images.  Do you know if any software (e.g. Image J plug-ins) has such a feature?

 

Thanks,

 

Lily




--
G. Esteban Fernandez, Ph.D.
Associate Director
Molecular Cytology Core Facility
University of Missouri
120 Bond Life Sciences Center
Columbia, MO 65211

http://www.biotech.missouri.edu/mcc/

573-882-4895
573-884-9395 fax