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Author Topic: Quantification of Immunofluorescence  (Read 9491 times)

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Offline wsnow

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Quantification of Immunofluorescence
« on: April 16, 2010, 12:25:38 PM »
Hi all,

I am trying to quantify some immunofluorescence from 2 different brain areas.  The protein of interest has a punctate distribution, so I have tried to use the Particle Analysis feature on ImageJ but am having some issues re: how to set the threshold appropriately.  This is key, as adjusting the threshold intensity manually gives you very different data points regarding the number and size of puncta. 

Has anyone had experience with this type of analysis and if so, do you have some parameters to follow when setting the threshold?

Many thanks,
Wanda Snow

Quantification of Immunofluorescence
« on: April 16, 2010, 12:25:38 PM »

Offline researcher4life

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Re: Quantification of Immunofluorescence
« Reply #1 on: April 19, 2010, 04:08:21 PM »
Hello,

I have used adobe CS4 software, there may be a newer version out now.  This was 3 years ago.

It will be in their photoshop package and is used for quantitative image analysis.  I used this software to quantify the intensity of FITC and Texas red stains for my projects.

Re: Quantification of Immunofluorescence
« Reply #1 on: April 19, 2010, 04:08:21 PM »