Quantum dots can be used to perform multicolor images with high fluorescent intensity and are of a nanosize suitable for lymphatic imaging via direct interstitial injection. Here simultaneous multicolor in vivo wavelength-resolved spectral fluorescence lymphangiography is shown using five quantum dots with similar physical sizes but different emission spectra. This allows noninvasive and simultaneous visualization of five separate lymphatic flows draining and may have implications for predicting the route of cancer metastasis into the lymph nodes.