I am using the LI-COR-Odessy system to image the membrane of western blots. Sometimes I can see blue markers covered on my bands. Maybe it is the indicator of overexposed bands. Does anyone know why it shows up? And how can I get rid of them? Thank you very much!
The blue markers that you are seeing on your Western blot membrane could be due to overexposure of the sample during imaging. These markers are often present on X-ray film as well and are indicative of overexposure of the sample.
To reduce the appearance of these blue markers, you can try the following:
Overall, the key to reducing the appearance of blue markers on your Western blot membrane is to optimize your experimental conditions and imaging settings to achieve a balance between signal intensity and background noise.
Hi Yuan Tian
It's important to understand that you cannot trust the results highlighted in blue, they've been highlighted for a reason and just turning off the option to display saturated bands is not the answer.
When bands are deemed "saturated" and highlighted in your image what it actually means is that the intensity of those bands has reached the upper limit of what can be captured by your equipment.
An example would be that if your equipment could handle an intensity of 10 units and your results actually had an intensity of 100 units, the image would stop at 10 because that's all your equipment can measure. You would then falsely believe that all those bands had an intensity value of 10 rather than whatever their "true" value is.
As mentioned above, the correct course of action would be to change your image capture settings (likely a shorter exposure time) to get all of your bands within the limit of detection for your LICOR device.
Only if you're not measuring the intensity of those highlighted bands is turning off saturation a reasonable course of action for rigorous results.
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