FBS Colloquia No.276Nuclear Dynamics Group

Tips for avoiding misinterpretation of fluorescence images

"We co-stained protein A, B and C with different colors, and then observed those localizations by microscope. The three proteins co-localized each other, suggesting that these proteins cooperatively function in the cell." Many biologists may have written this kind of description in their paper. But wait a moment! Is this discussion scientifically correct? In most cases, this type of discussion is incorrect or it is difficult for readers to assess its correctness from the description in the paper. This misinterpretation of the data results from a chromatic aberration of the objective lens: an unexpected chromatic shift is introduced into your image when you observe an object by microscope. The aberrations of the objective lens are becoming a severe problem for the high-resolution microscope, especially super-resolution microscopes. In this seminar, I will explain various aberrations which objective lenses have, and introduce some Tips to correctly assess your fluorescence images.