Published online by Cambridge University Press: 14 March 2018
Fluorescence imaging of brains from older animals, particularly primates, can be hindered by the presence of lipofuscin, a strongly fluorescent pigment that accumulates with age, primarily in neuronal cells. Lipofuscin has a broad emission spectrum that overlaps emission of most fluorescent labels, is present throughout the brain and other tissues, and is very hard to quench chemically (Schnella et at.. Journal of Histochemistry and Cytochemistry, Vol. 47, 719-730, 1999). A more convenient way to eliminate lipofuscin from fluorescence images is the use image arithmetic to subtract its signal. This method takes advantage of lipofuscin's broad emission spectrum that makes it appear in all fluorescence channels.