The use of quantum dots for analysis of chick CAM vasculature

Quantum dots (QDs) are fluorescent semiconductor nanocrystals that possess a number of superior fluorescent properties compared to more established organic dyes and fluorescent proteins. As a result, QDs are being studied for use in a wide range of biological applications. We have examined QDs for one such application, visualization of blood vessels of the chick chorioallantoic membrane (CAM), a popular model for studying various aspects of blood vessel development including angiogenesis. Intravitally injected QDs were found to be biocompatible and were kept in circulation over the course of 4 days without any observed deleterious effects. QD vascular residence time was tunable through QD surface chemistry modification. We also found that use of QDs with higher emission wavelengths (>655 nm) virtually eliminated all chick-derived autofluorescence and improved depth-of-field imaging. QDs were compared to FITC-dextrans, a fluorescent dye commonly used for imaging CAM vessels. QDs were found to image vessels as well as or better than FITC-dextrans at 2-3 orders of magnitude lower concentration. We also demonstrated that QDs are fixable with low fluorescence loss and thus can be used in conjunction with histological processing for further sample analysis.