How to split Green/Red/Yellow

Dear Sir:

I have a image containing green cells with green fluorescent protein  
(GFP), red cells with red fluorescent protein (RFP), and yellow cells  
with both (GFP and RFP).
I would like to measure the number/area of each of green, red, and  
yellow cells.
By 'RGB Split', I can separate green cells and red cells.
However, I do not know how to get a image containing only yellow cells.
I wonder if you could tell me how to analyze these three different  
color particles separately.

I am looking forward to hearing from you.

Sincerely yours,

Haruo Suzuki
--------------------------------------
Protect Your PC Now ! -- 0 Yen Security Campaign
http://pr.mail.yahoo.co.jp/security/

[Ricardo Henriques-2]

Hi,

Do the RGB split then go to Process => Image Calculator, there you can
use the AND operation to get an image with the pixels that colocalize in
both images.

Cheers,

Ricardo Henriques

[hidden email] wrote:

> Dear Sir:
>
> I have a image containing green cells with green fluorescent protein
> (GFP), red cells with red fluorescent protein (RFP), and yellow cells
> with both (GFP and RFP).
> I would like to measure the number/area of each of green, red, and
> yellow cells.
> By 'RGB Split', I can separate green cells and red cells.
> However, I do not know how to get a image containing only yellow cells.
> I wonder if you could tell me how to analyze these three different
> color particles separately.
>
> I am looking forward to hearing from you.
>
> Sincerely yours,
>
> Haruo Suzuki
> --------------------------------------
> Protect Your PC Now ! -- 0 Yen Security Campaign
> http://pr.mail.yahoo.co.jp/security/
>

Thank you very much for your advice.

I would like to ask additional questions:
Is it possible to separate red and yellow cells which are located on  
same position on x- and y-axis but different position on z-axis?
Is it possible to calculate 'volume' of each colors (red, green, and  
yellow) instead of 'area'?

Sincerely yours,

Haruo Suzuki

On 2006/11/07, at 1:27, Ricardo Henriques wrote:

> Hi,
>
> Do the RGB split then go to Process => Image Calculator, there you  
> can use the AND operation to get an image with the pixels that  
> colocalize in both images.
>
> Cheers,
>
> Ricardo Henriques
>
> [hidden email] wrote:
>> Dear Sir:
>>
>> I have a image containing green cells with green fluorescent  
>> protein (GFP), red cells with red fluorescent protein (RFP), and  
>> yellow cells with both (GFP and RFP).
>> I would like to measure the number/area of each of green, red, and  
>> yellow cells.
>> By 'RGB Split', I can separate green cells and red cells.
>> However, I do not know how to get a image containing only yellow  
>> cells.
>> I wonder if you could tell me how to analyze these three different  
>> color particles separately.
>>
>> I am looking forward to hearing from you.
>>
>> Sincerely yours,
>>
>> Haruo Suzuki
>> --------------------------------------
>> Protect Your PC Now ! -- 0 Yen Security Campaign
>> http://pr.mail.yahoo.co.jp/security/
>>

--------------------------------------
Protect Your PC Now ! -- 0 Yen Security Campaign
http://pr.mail.yahoo.co.jp/security/

[Sergio Caballero]

You're better off using Intensity Correlation Analysis plugin
(http://www.uhnresearch.ca/facilities/wcif/software/Plugins/ICA.html) which
will give you much more accurate results. Be sure to read the PDF
documentation. As far as calculating volume, measure the area of
colocalization for each z-slice, sum them, and then multiply the sum by
z-thickness to get a near approximation of actual volume. The smaller your
z-thickness the closer to actual volume you'll get (it's a derivative after
all).

-----Original Message-----
From: ImageJ Interest Group [mailto:[hidden email]] On Behalf Of
[hidden email]
Sent: Tuesday, November 07, 2006 1:01 PM
To: [hidden email]
Subject: Re: How to split Green/Red/Yellow

Thank you very much for your advice.

I would like to ask additional questions:
Is it possible to separate red and yellow cells which are located on  
same position on x- and y-axis but different position on z-axis?
Is it possible to calculate 'volume' of each colors (red, green, and  
yellow) instead of 'area'?

Sincerely yours,

Haruo Suzuki

On 2006/11/07, at 1:27, Ricardo Henriques wrote:

> Hi,
>
> Do the RGB split then go to Process => Image Calculator, there you  
> can use the AND operation to get an image with the pixels that  
> colocalize in both images.
>
> Cheers,
>
> Ricardo Henriques
>
> [hidden email] wrote:
>> Dear Sir:
>>
>> I have a image containing green cells with green fluorescent  
>> protein (GFP), red cells with red fluorescent protein (RFP), and  
>> yellow cells with both (GFP and RFP).
>> I would like to measure the number/area of each of green, red, and  
>> yellow cells.
>> By 'RGB Split', I can separate green cells and red cells.
>> However, I do not know how to get a image containing only yellow  
>> cells.
>> I wonder if you could tell me how to analyze these three different  
>> color particles separately.
>>
>> I am looking forward to hearing from you.
>>
>> Sincerely yours,
>>
>> Haruo Suzuki
>> --------------------------------------
>> Protect Your PC Now ! -- 0 Yen Security Campaign
>> http://pr.mail.yahoo.co.jp/security/
>>

--------------------------------------
Protect Your PC Now ! -- 0 Yen Security Campaign
http://pr.mail.yahoo.co.jp/security/

[Adam Rodaway]

Hi Haruo,
an alternative way to do this is to use the 'RG2B Colocalization'
plugin. This will put your yellow cells in the blue channel (they will
also still be in the red and green channels).
best wishes
Adam
On 6 Nov 2006, at 23:34, [hidden email] wrote:

> Dear Sir:
>
> I have a image containing green cells with green fluorescent protein
> (GFP), red cells with red fluorescent protein (RFP), and yellow cells
> with both (GFP and RFP).
> I would like to measure the number/area of each of green, red, and
> yellow cells.
> By 'RGB Split', I can separate green cells and red cells.
> However, I do not know how to get a image containing only yellow cells.
> I wonder if you could tell me how to analyze these three different
> color particles separately.
>
> I am looking forward to hearing from you.
>
> Sincerely yours,
>
> Haruo Suzuki
> --------------------------------------
> Protect Your PC Now ! -- 0 Yen Security Campaign
> http://pr.mail.yahoo.co.jp/security/
>
>

Dr Adam Rodaway
http://www.kcl.ac.uk/depsta/biomedical/randall/arodaway.html

King's College London
The Randall Centre
3rd Floor North
New Hunt's House
Guy's Campus
London SE1 1UL

tel. office [phone]
tel. lab. [phone]
fax [phone]
tel. mobile [phone]

[Adam Rodaway]

'RG2B Colocalization' works on individual slices so will only show
colocalization in the same Z position if applied to stack before you
z-project.
regards
Adam
On 7 Nov 2006, at 18:01, [hidden email] wrote:

> Thank you very much for your advice.
>
> I would like to ask additional questions:
> Is it possible to separate red and yellow cells which are located on
> same position on x- and y-axis but different position on z-axis?
> Is it possible to calculate 'volume' of each colors (red, green, and
> yellow) instead of 'area'?
>
> Sincerely yours,
>
> Haruo Suzuki
>
> On 2006/11/07, at 1:27, Ricardo Henriques wrote:
>
>> Hi,
>>
>> Do the RGB split then go to Process => Image Calculator, there you
>> can use the AND operation to get an image with the pixels that
>> colocalize in both images.
>>
>> Cheers,
>>
>> Ricardo Henriques
>>
>> [hidden email] wrote:
>>> Dear Sir:
>>>
>>> I have a image containing green cells with green fluorescent protein
>>> (GFP), red cells with red fluorescent protein (RFP), and yellow
>>> cells with both (GFP and RFP).
>>> I would like to measure the number/area of each of green, red, and
>>> yellow cells.
>>> By 'RGB Split', I can separate green cells and red cells.
>>> However, I do not know how to get a image containing only yellow
>>> cells.
>>> I wonder if you could tell me how to analyze these three different
>>> color particles separately.
>>>
>>> I am looking forward to hearing from you.
>>>
>>> Sincerely yours,
>>>
>>> Haruo Suzuki
>>> --------------------------------------
>>> Protect Your PC Now ! -- 0 Yen Security Campaign
>>> http://pr.mail.yahoo.co.jp/security/
>>>
>
> --------------------------------------
> Protect Your PC Now ! -- 0 Yen Security Campaign
> http://pr.mail.yahoo.co.jp/security/
>
>

Dr Adam Rodaway
http://www.kcl.ac.uk/depsta/biomedical/randall/arodaway.html

King's College London
The Randall Centre
3rd Floor North
New Hunt's House
Guy's Campus
London SE1 1UL

tel. office [phone]
tel. lab. [phone]
fax [phone]
tel. mobile [phone]