Area fraction ... is it the same as " % of positive cells " ?

[Amal Kamal]

Dear All,
  Right now ... I am trying to use ImageJ to calculate the % of positive cells (stained using immunohistochemistry technique) ... I have read an article stating that we can use ImageJ to calculate the "area fraction" ... but I can't get it whether it is equivalent to " % of positive cells "...?
Can anyone help ?
Thanks alot.
Best regards,
Amal Kamal

[carlo bianco]

Dear Amal,
it  depends on what IHC stains: nuclei or cytoplasm or membrane?
I found a robust software online available also as a plugin: immunoratio
http://153.1.200.58:8080/immunoratio/
of course area fraction is measured area
%of cells depends on anisocytosis I think,
I hope my suggestion is useful,
best wishes
carlo bianco-bologna




________________________________
 Da: Amal Kamal <[hidden email]>
A: [hidden email]
Inviato: Sabato 26 Gennaio 2013 18:30
Oggetto: Area fraction ... is that the same of % of positive cells ?
 
Dear All,
  Right now ... I am trying to use ImageJ to calculate the % of positive
cells (stained using immunohistochemistry technique) ... I have read an
article stating that we can use ImageJ to calculate the "area fraction" ...
but I can't get it whether it is equivalent to " % of positive cells "...?
Can anyone help ?
Thanks alot.
Best regards,
Amal Kamal



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[Amal Kamal]

Dear Carlo,
Thanks alot for your kind reply.
Actually, I have images of both cytoplasmic markers ... and others of nuclear ones ... the link you provided above is used for quantitation of nuclear DAB stain ... right ?
Thanks alot for your help.
Best regards,
Amal Kamal

[carlo bianco]

Dear Amal,
the plugin I suggested  you is developed for a purpose,
but in my opinion other  dab nucelar positivity with hematoxilin counterstain are
handable, maybe before measures the coorrelation between manual counting and
plugin counting should be analyzed.
for the cytoplasm I think depends on shape of the cells and on
tissue you aer working. I know only immunomembrane, maybe
with a saturation step before acquisition of photo, the membranous stain can
be enhanced and measured as an indicator of cytoplasmic stain. This must be tried.
I hope this is useful.,
best wishes for your trial
carlo   bologna





________________________________
 Da: Amal Kamal <[hidden email]>
A: [hidden email]
Inviato: Sabato 26 Gennaio 2013 21:55
Oggetto: Re: Area fraction ... is that the same of % of positive cells ?
 
Dear Carlo,
Thanks alot for your kind reply.
Actually, I have images of both cytoplasmic markers ... and others of
nuclear ones ... the link you provided above is used for quantitation of
nuclear DAB stain ... right ?
Thanks alot for your help.
Best regards,
Amal Kamal



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[Amal Kamal]

Thanks alot Carlo for your help.
Best regards,
Amal Kamal

[Stoyan Pavlov]

Dear Amal,
The simple answer is no. Positive area fraction is not directly equivalent
to percentage of positive cells. The mean area fraction is actually equal
to the positive volume fraction of the sampled volume. If you know the
latter you can directly calculate the positive volume.
However depending on the cells you are trying to evaluate
 you can make an estimate of the cell number (percentage). If you are sure
that the measured cells react only with changes in numbers than the change
in area (volumetric) fraction will be equivalent to
changes in the cell numbers. If the cells react with changes in form, shape
and numbers than your task is a bit more complicated. If absolute cell
numbers is not your goal ( but only general cellular reaction
) you can compare the fractions directly. If the result of interest are
cell numbers than you can still calculate an estimate if you know the mean
volume of the corresponding cells. However in such case the best aproach
would be unbiased stereological counting.
Of course all this explanations count only if you are staining the
cytoplasm. If you're specifically staining the cell nuclei of course you
can measure the fraction of positive cells in a section, but this is by far
not the same as the positive cell fraction in the sampled  volume and there
is no easy way to calculate an estimate if you aren't using unbiased
stereology.

Stoyan Pavlov



2013/1/27 Amal Kamal <[hidden email]>

> Thanks alot Carlo for your help.
> Best regards,
> Amal Kamal
>
>
>
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> View this message in context:
> http://imagej.1557.n6.nabble.com/Area-fraction-is-it-the-same-as-of-positive-cells-tp5001554p5001560.html
> Sent from the ImageJ mailing list archive at Nabble.com.
>
> --
> ImageJ mailing list: http://imagej.nih.gov/ij/list.html
>

--
Dr. Stoyan P. Pavlov, MD, PhD
Departament of Anatomy, Histology and Embryology
Medical University "Prof. Dr. Paraskev Stoyanov", Varna
Prof. Marin Drinov Str.55
9002 Varna
 Bulgaria
Tel: +359 (0) 52 - 677 - 086
e-mail: [hidden email]

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[Amal Kamal]

Dear Stoyan
Thanks alot for your help. Actually, I have IHC images with cytoplasmic markers... and yes, the antibody is supposed to react with the specific protein in the cytoplasm and my goal is to calculate the % of positive cells ...