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Re: Creating Pseudocolor Images with more than 3 channels

Posted by Wayne Rasband on Jun 06, 2008; 5:22pm
URL: http://imagej.273.s1.nabble.com/Creating-Pseudocolor-Images-with-more-than-3-channels-tp3696013p3696014.html

> How do I create a pseudo-color images with more than 3 colors. I have a
> set of images with 5 channels with different bio-markers, and would
> like
> to specify the color for each marker. For example, blue color for DAPI,
> green color for cytoplamic marker, orange color for target protein.

    1. Create a stack containing the 5 images
    2. Use Image>Color>Make Composite to create a 5 channel composite
image
    3. Press shift-z (Image>Hyperstack>Channels) to open the Channels
window
    4. Select "Color" in the Channels window
    5. Use the slider under the composite image to select the DAPI
channel
    6. Select Image>Lookup Tables>Blue
    7. Select the cytoplamic marker channel
    8. Select Image>Lookup Tables>Green
    9. Select the target protein channel
    10. Open the LUT Editor (Image>Color>Edit LUT)
    11. Create an orange LUT (drag across all 256 entries then set
red=255, green=~160, blue=0)
    12. Add LUTs to the other 2 channels
    13. Select "Composite" in the Channels window
    14. Use the B&C tool (press shift-c to open) to adjust each of the
channels
    15. Use the Channels tool to enable/disable channels

For practice, open the 5 channel File>Open Samples>Neuron image, which
was added in v1.41d.

> In addition, is there any automated registration plugin or tools for
> the
> alignment for these images? Any suggestion for registration of images
> is
> highly appreciated.

There is the StackReg plugin at
<http://bigwww.epfl.ch/thevenaz/stackreg/>.

-wayne


>
> Thanks,
>
> Zhengyu
>
>> g Global Research Center
>>
>> ______________________________________________________________________
>>
>>           Zhengyu Pang, Ph.D.
> Bioanalytics and Protein Science Laboratory
> Biosciences Global Technology Organization
>> One Research Circle, K1 5B37A
>> Niskayuna, NY12309
>> * [hidden email]
>> *(518) 387-4015
>>
>>
>