I have read some papers where the authors showed that parcentage of cells harboring parkin in mitochondria (e.g. "Parkin is recruited selectively to impaired mitochondria and promotes their autophagy" and its addendum in the journal Autophagy). These articles are very significant input to mitochondrial biology as well as autophagy. I am very much curious about the co-localization experiment they have performed. I like to ask if anybody can help me understanding method of co-localization they have used. I have already asked them.
The basically used the FLIP assay where they overexpressed YFP-Parkin and DsRed-Mito in a cell, find a spot that has the mitochondrial signal, photobleach the two channels, and do time lapse and track level of YFP and DsRed re-entering that region. You should also note that they did a lot of confocal immunofluorescence showing in the z-axis that the parkin signal overlaps with the mitochondrial signal.
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