Spectral unmixing

[Amolkumar Karwa]

Hello All,

I'm using Quantum Dots for fluorescence imaging. These three QDs have their emission maximum at 605,655 and 705nm with a little bleed over. I want to spectrally resolve these Quantum Dots using the plugins provided but I don't have any control image with single QDs.Is it still possible to differentiate between these 3 QDs?

Thanks,

Amol

[Joachim Walter]

Sure, if you have a sample, in which the three types of Quantum Dots are
separated in space, you can use the "Spectral Unmixing" Plugins
(http://rsb.info.nih.gov/ij/plugins/spectral-unmixing.html) to do
that. The PDF-Documentation
(http://rsb.info.nih.gov/ij/plugins/docs/SpectralUnmixing.pdf) explains
how to do that. Your case is
described as "case 1" in the documentation.

Joachim

Amolkumar Karwa wrote:

> Hello All,
>
> I'm using Quantum Dots for fluorescence imaging. These three QDs have
> their emission maximum at 605,655 and 705nm with a little bleed over.
> I want to spectrally resolve these Quantum Dots using the plugins
> provided but I don't have any control image with single QDs.Is it
> still possible to differentiate between these 3 QDs?
>
> Thanks,
>
> Amol

--
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   Dr. Joachim Walter
   Till I.D. GmbH
   c\o BioImaging Zentrum
   Großhaderner Str. 2
   82152 Martinsried, Germany

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   Fax:  [phone]

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[etanha]

Dear Dr.Walter

I am an MSc student of medical physics. I coudn't send directly Email for you.

My thesis is about multispectral imaging. I sawe your web page: "Spectral Unmixing Plugins". I am very interested in your work.

I have a problem with unmixing images.pleas explain for me the linear  spectral unmixing algorithm. I have used SVD to decompose the images. I did so and finally obtained three matrices. after inverting them I multiplied those matrices into the observed image from left to get the unmixed image. But the dimensions of the resulted matrix and the observed image are not the same. I used MATLAB to do all of them and 2 flourophor for my work.
I highly appreciate if you help me to understand what is wrong in my procedure.

Thanks in advance
Sincerely yours

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Ebrahim Najafzadeh
Tehran University of Medical Sciences