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We have developed two tools that can be used to quantify the intensity of one fluorescent marker that overlays another fluorescent marker. We have used these to quantify the buildup of actin filaments on mitochondria, which we refer to as ADA (acute damage-induced actin, described in PMID 35290799). However, these methods can be used to quantify any change in fluorescence in specific areas of the cell (as long as that area is fluorescently labeled itself).
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