Measuring Callose area

Hey,
I usually use FIJI to analyze confocal images to determine the area of callose. The images are pretty simple with fluorescence sometimes outlining the cell membrane slightly and stronger intensity at the plasmodesmata where callose accumulates. This appears as a circular shape with strong fluorescence. Any suggestions of how to process the confocal image and how use CellProfiler to identify the object and obtain measurements.
Here is an example of how I use FIJI to identify and measure the area.
(upload://5mM0vgLANXsLqXJGSnvY8ZeisCV.png)
Thanks!

@hvariz

Please attach an original image file … this will help us better help you.

Hi @hvariz,

I think you might have to attach a sample raw image. Also check if any these example pipelines will of any help to start with.

Regards,
Lakshmi
Fujifilm Wako Automation (Consultant)
www.wakoautomation.com
For CellProfiler training or optimised pipeline write to,
lakshmi.balasubramanian.contractor@fujifilm.com

Read more on our site.
Yokogawa CV8000 - The Ultimate in Confocal HCS
https://www.wakoautomation.com/products/yokogawa-high-content-imaging

5.tif (1.0 MB)
Also here is a raw image in tiff format
Thanks

Hi @lakshmi Thanks for the advice

Hi @hvariz,

You are welcome!!!Here is the quick pipeline i tried with you image. This must give you a quick start and more optimisation might give your expected results more precisely. To export the measurements to an excel sheet you could add “ExportToSpreadmodule” while you are doing the batch process. PFA pipeline & segmented overlay

screenshot.pdf (1.1 MB)
callose_trail.cpproj (759.4 KB)

Hope this helps.

Regards,
Lakshmi
Fujifilm Wako Automation (Consultant)
www.wakoautomation.com
For CellProfiler training or optimised pipeline write to,
lakshmi.balasubramanian.contractor@fujifilm.com

Read more on our site.
Yokogawa CV8000 - The Ultimate in Confocal HCS
https://www.wakoautomation.com/products/yokogawa-high-content-imaging

2 Likes

Wow that’s amazing thank you so much!