MCD BIO 165A Lecture Notes - Lecture 2: Total Internal Reflection, Total Internal Reflection Fluorescence Microscope, Deconvolution
11 Feb 2018
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Question: whether plasma membrane proteins are distributed homogeneously or not. Integral proteins - transmembrane plasma membrane proteins: peripheral proteins - noncovalent interactions (not mentioned in this paper) Lipid anchor proteins - covalently linked to lipids. Is the distribution of proteins into specific micro-domains important for their function: dynamics of proteins. Tools: tirf microscopy, resolution improved through deconvolution and sim, deconvolution - computationally remove distortion. Sim - super resolution technique to transcend the point spread problem. Purpose of figure: compare two different methods of tirf imaging. Determined that tirf sim and deconvolution were similar enough. A: different categories of proteins randomly chosen for the study, several transmembrane proteins and several lipid anchors, 42 randomly-chosen membrane proteins were tagged with gfp. Limitations: can affect function, folding, or localization of protein: advantages: don"t need to use a bunch of different antibodies to detect; can visualize in a live cell.
