I have done IF with Abcam antibody, but the image that I get is not look like as the Abcam review on the same anitbody.
My image (attached red image) is kind blush-like or blur color?, but from the review is look dot-like color (green image, http://www.abcam.com/calcineurin-a-antibody-ab3673.html/reviews/32402). What are factors that make these two image so different? and Is these a way to improve my image to be dot like as closed as the review image?
Thank you
Cheerio,
Are you following the protocol previously used exactly? is there a difference in cell type? fixation conditions?
Hello Asa,
First of all I am surprised that why u want a dot like image? Infact your image is better than the reference image. OK, as a hardcore microscopy researcher let me tell you, the green dot like image is at higher magnification and the expression of antibody or staining intensity is poor, so they people have set the LUTs to increase the expression of green color which has resulted in dot like expression. It means the pixels of image have distorted in this high magnification and LUT enhancement. Thats the reason for image to be dotted. If u want to make your image like this, you can also play with LUTs , (a left hand movement from extreme point on RHS in red channel) and increase magnification, you will get the desired image. But according to me, your image is better, for improvement just increase the magnification and m sure you are gonna get a better image than the reference one..
Hope it will be helpful for you...!
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