Carbon nanotubes are promising new materials for molecular delivery in biological systems. The long-term fate of nanotubes intravenously injected into animals in vivo is currently unknown, an issue critical to potential clinical applications of these materials. Here, using the intrinsic Raman spectroscopic signatures of singlewalled carbon nanotubes (SWNTs), we measured the blood circulation of intravenously injected SWNTs and detect SWNTs in various organs and tissues of mice ex vivo over a period of three months. Functionalization of SWNTs by branched polyethyleneglycol (PEG) chains was developed, enabling thus far the longest SWNT blood circulation up to 1 day, relatively low uptake in the reticuloendothelial system (RES), and near-complete clearance from the main organs in Ϸ2 months. Raman spectroscopy detected SWNT in the intestine, feces, kidney, and bladder of mice, suggesting excretion and clearance of SWNTs from mice via the biliary and renal pathways. No toxic side effect of SWNTs to mice was observed in necropsy, histology, and blood chemistry measurements. These findings pave the way to future biomedical applications of carbon nanotubes.biodistribution ͉ blood circulation ͉ nanoparticles ͉ excretion ͉ toxicity T he utilization of novel nanomaterials for biological and biomedical applications has been an active and exciting direction of research in recent years (1-3). A wide range of nanomaterials, such as nanoparticles (4 -7), nanorods (8), nanowires (9), and nanotubes (10-12) have been investigated for their potential clinical applications in diagnosis and therapeutic treatment of diseases. The interesting structural, chemical, electrical, and optical properties of carbon nanotubes (13,14), when used in biological and medical settings, may bring new opportunities to biological detection, imaging, and therapy with high performance and efficacy. Carbon nanotube-based intercellular molecular delivery vehicles have been developed for intracellular gene (15-17) and drug delivery in vitro (18,19). Recently, research began to investigate the behavior of carbon nanotubes in animal bodies in vivo (20,21), including the finding of lack of toxicity of well PEGylated single-walled carbon nanotubes (SWNTs) in mice in a pilot study (M. L. Schipper, N. NakayamaRatchford, C.D., N. W. S. Kam, P. Chu, Z.L., X. Sun, L. C. Cork, H.D., and S. S. Gambhir, unpublished data).The biodistribution of intravenously injected carbon nanotubes in mice has been studied by using radiolabeling and isotope ratio mass spectroscopy methods (20,22). Promising result of efficient targeted tumor accumulation in vivo has been obtained by conjugating a ligand peptide to nanotubes to recognize receptors on the surface of tumor cells (20), suggesting high potential of nanotube-based drug delivery vehicles for cancer therapy. However, an important unaddressed question for carbon nanotubes and various nanomaterials in general in biomedical applications is their long term fate in vivo. It is known that most nanomaterials tend to exhibit high upt...