Monitoring lysosomal activity in nanoparticle-treated cells.
Certain nanoparticles have been shown to accumulate within lysosome and hence may cause lysosomal pathologies such as phospholipidosis, lysosomal overload, and autophagy. This chapter describes a method for evaluation of lysosomal activity in porcine kidney cells (LLC-PK1) after exposure to nanoparticles. This method uses the accumulation of a cationic fluorescent dye (LysoTracker Red) in acidic cellular compartments as an indicator of total lysosome content. The lysotracker signal is normalized to the signal from a thiol-reactive dye which is proportional to the total number of viable cells.
Nanotechnology Characterization Laboratory, Advanced Technology Program, SAIC-Frederick, Inc., National Cancer Institute at Frederick, Frederick, MD, USA.
Barry W
BW
Neun
Stephan T
ST
Stern
D000588:Amines / Q000032:analysis / Q000378:metabolism; D000818:Animals; D001343:Autophagy / Q000187:drug effects* / Q000502:physiology; D002470:Cell Survival / Q000187:drug effects; D004847:Epithelial Cells / Q000187:drug effects; D005452:Fluoresceins / Q000378:metabolism; D005456:Fluorescent Dyes / Q000032:analysis* / Q000378:metabolism; D007668:Kidney / Q000187:drug effects; D018374:LLC-PK1 Cells; D008247:Lysosomes / Q000378:metabolism*; D053758:Nanoparticles / Q000633:toxicity*; D015398:Signal Transduction; D013552:Swine
D000588:Amines; D005452:Fluoresceins; D005456:Fluorescent Dyes; C493330:Red DND-99; C069306:5-chloromethylfluorescein