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Re: threshold

Posted by Sarah Mohamed on Oct 18, 2014; 6:18pm
URL: http://imagej.273.s1.nabble.com/threshold-tp5009636p5010103.html

Dear Jacqui,
thanks for your reply, i appreciate it.

In your opinion, which stain can be used (to avoid the DAB issue with quantification) to quantify a marker in treated and untreated samples?


Kind Regards,
Sarah M. Mosaad, M.Sc.
Department of Pharmacology & Toxicology,
School of Pharmacy,
Suez Canal University, Egypt.



On Sunday, October 12, 2014 10:06 PM, Jacqui Ross <[hidden email]> wrote:
 


Hi Sarah,



Just noticed that no one seems to have replied to your question. This is probably because we aren't sure what "results" you are referring to and what you mean by "threshold".



If you are measuring areas, then it's OK to use some processing to make the segmentation easier as long as you don't start to bring up some areas which were not really stained.



If you were measuring intensity for fluorescence images, then you shouldn't use any processing which changes the pixel values.



However, we know that measuring intensity/density of DAB staining is not acceptable. This has been discussed many times on the listserver.



Please read the previous thread here: http://imagej.1557.x6.nabble.com/DAB-quantification-td5006159.html

<http://imagej.1557.x6.nabble.com/DAB-quantification-td5006159.html> in particular the comments by Gabriel Landini in reference to DAB quantification.



Kind regards,



Jacqui



Jacqueline Ross

Biomedical Imaging Microscopist

Biomedical Imaging Research Unit

School of Medical Sciences

Faculty of Medical & Health Sciences

The University of Auckland

Private Bag 92019

Auckland 1142, NEW ZEALAND



Tel: 64 9 923 7438

Fax: 64 9 373 7484



http://www.fmhs.auckland.ac.nz/sms/biru/





-----Original Message-----

From: ImageJ Interest Group [mailto:[hidden email]] On Behalf Of Sarah Mohamed

Sent: Tuesday, 16 September 2014 10:15 p.m.

To: [hidden email]<mailto:[hidden email]>

Subject: threshold



Hi,

Can I manipulate with the image brightness before analysis using threshold?





I am trying to determine brown area stained with DAB using image J, i tried to use threshold and color deconvolution. my images a dark can i use photoshop to adjust background or adjust brightness (using levels or curves) ? or that will change the results?





Regards,Sarah M. Mosaad, M.Sc.



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