Re: Quantifying Stained Retina

Posted by NatyC on
URL: http://imagej.273.s1.nabble.com/Quantifying-Stained-Retina-tp5000880p5001009.html

Dear Karsten,

How would you recommend measuring these ROI?

I've tested out the basic Analyze>Set Measurements, Analyze>
Measurement on a Split Channel (RGB). With this process I measured the
area,integrated density, and mean gray density of the Split Channel
Blue image.

The issue with this analysis is that there is no true distiction in
the amount of "Blue"
Rather, all of the values ended up to be the total amount of pixels
that compose the image.

Natalia

On 11/28/12, Karsten <[hidden email]> wrote:

> Hi,
> this is an interesting thread with many very differing layers of
> understanding of images!
>
> What about measuring the "amount" of blue, which is IMHU the total
> extinction of the blue channel, in a selected region, e.g. a layer or a part
> of it? For standardization the area of the region should be measured too, or
> if the nuclei are of interest, better the area of the nuclei inside the
> selected region.
>
> ... and so on, step by step a real measurement scheme will be established.
>
> For reliable measurements lots of questions will arise! If "the iron"
> stained by "Pearls Blue" is in any sense stoichiometric perhaps images
> should be sampled as pure intensity image using the appropriate filter!
>
> However, I would recommend to measure, as the mentor ask for, and look at
> the results.
> Also mentors are still learning!
>
> Regards
> Karsten
>
> Am 28.11.2012 um 21:55 schrieb Natalia Chacon:
>
>> On 11/28/12, Natalia Chacon <[hidden email]> wrote:
>>> Dear Mr. Gabriel,
>>>
>>> I've enclosed another image that seems to contain more iron in the
>>> sample.
>>> Also, is it still possible to use the Color Deconvolution to get the
>>> data my mentor desires?
>>>
>>> Thanks,
>>> Natalia
>>>
>>> On 11/20/12, Gabriel Landini <[hidden email]> wrote:
>>>> On Tuesday 20 Nov 2012 14:55:39 Natalia Chacon wrote:
>>>>> Rob, my mentor told me she used a stain called "Pearls Blue"
>>>>>
>>>>> Joel, I'm interested in quantifiying most of the stainings visible.
>>>>> (the background staining I don't desire to quantify.)
>>>>>
>>>>> What you're stating though does correlate with what my mentor has
>>>>> informed
>>>>> me. That the onl nuclei would be the most dense.  This would be due to
>>>>> the
>>>>> way the protien settles.
>>>>
>>>> This is what Prussian, Pearls or Berlin blue looks like:
>>>>
>>>> http://www.jichi.ac.jp/pathology/swfu/d/fe-10.jpeg
>>>>
>>>> Your image looks like all contrast stain to me. So you might not have
>>>> any
>>>> iron
>>>> compounds there or it is so faint that the contrast stain prevails.
>>>>
>>>> With Berlin blue, you can *demonstrate* that some iron compound is
>>>> there,
>>>> but
>>>> I am not sure if it can be used to quantify their quantity. Here it
>>>> says
>>>> it
>>>>
>>>> cannot, so I would be careful how to interpret the intensity:
>>>>
>>>> http://stainsfile.info/StainsFile/stain/pigment/perls.htm
>>>>
>>>> Regards
>>>>
>>>> Gabriel
>>>>
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>>>>
>>>
>>
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