Abstract
The compound 1,1’-dioctadecyl-3,3,3’,3’-tetramethylindocarbocyanine perchlorate (Dil) is a fluorescent molecule with diffusion properties similar to those of phospholipids. Intramembranous (“lateral”) diffusion of Dil is preserved in aldehyde-fixed tissues and has been used to label neuronal pathways. We found that Dil-label, when illuminated (“photo-converted”) in the presence of diaminobenzidine (DAB), forms an electron-dense reaction product that is suitable for EM analysis. We show that the Dil molecule has a surprising diffusion ability and differential affinitiy to internal membranes after retrograde diffusion along neurites in fixed tissue. This finding may have implications for normal diffusion of phospholipids along intracellular pathways in vivo.Two 20-day old White Leghorn chick embryos were anesthetized and perfused intracardially with 0.1M phosphate buffer (PB, pH 7.4) followed by perfusion with 2% paraformaldehyde and 1% glutaraldehyde in PB.
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Schedule a callMore From: Proceedings, annual meeting, Electron Microscopy Society of America
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